LOCUS       V032979                 9368 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032979
VERSION     V032979
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(join(9282..9368,1..18))
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     492..508
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     LTR             751..931
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    975..1100
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1593..1826
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             2010..2054
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2315..2432
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2522..2725
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2799..2843
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             2877..2900
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2904..2927
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2937..3050
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             3132..4754
                     /label="Ugt8a(NM_011674)"
                     /note="Ugt8a(NM_011674)"
                     /gene="Ugt8a"
     CDS             4761..4784
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             4788..4811
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4855..5066
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5079..5347
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5374..5973
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5980..6568
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6642..6875
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6947..7068
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7087..7222
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7260..7276)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7284..7300
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7308..7338)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7353..7374
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7662..8250)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8421..9281)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
ORIGIN
        1 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
       61 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
      121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      241 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
      361 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      421 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
      481 gtcacgacgt tgtaaaacga cggccagtgc caagctgacg cgtgtagtct tatgcaatac
      541 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
      601 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
      661 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
      721 tatttaagtg cctagctcga tacaataaac gggtctctct ggttagacca gatctgagcc
      781 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      841 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      901 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ctgaaagcga
      961 aagggaaacc agagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1021 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1081 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1141 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1201 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1261 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1321 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1381 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1441 ccactgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     1501 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     1561 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     1621 gcagcaggaa gcactatggg cgcagcctca atgacgctga cggtacaggc cagacaatta
     1681 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     1741 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     1801 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1861 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gattggaatc
     1921 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1981 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2041 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2101 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2161 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2221 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2281 ccattcgatt agtgaacgga tctcgacggt taacttttaa aagaaaaggg gggattgggg
     2341 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2401 tacaaaaaca aattacaaaa attcaaaatt ttatcgatac tagtattatg cccagtacat
     2461 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2521 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     2581 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     2641 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     2701 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     2761 tccacgctgt tttgacctcc atagaagatt ctagaatgaa agaaaccgct gctgctaaat
     2821 tcgaacgcca gcacatggac agcggagccg gcgcaggagc cggcgctgac gcgcctgact
     2881 acaaagacga tgacgacaag ggagattaca aggatgacga tgacaaaggc gcgtcgatgg
     2941 acgagaagac caccggctgg cggggcggcc acgtggtgga gggcctggcc ggcgagctgg
     3001 agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc tccggcggct
     3061 gcaagctggg cggcgcgcct gaattctgca gtcgacggta ccgcgggccc atcgctcgat
     3121 ctagagctag catgaagtct tacactccat atttcatgct cctgtggagt gctgttggaa
     3181 tagcaagggc tgctaaaatc atcattgtgc cgccaattat gtttgaaagc catttgtaca
     3241 ttttcaagac actagcatcg gccttgcacg agagaggcca tcacaccgtg ctcctcctct
     3301 cagaaggcag agacattgcc ccatctaatc actacagcct ccagcggtac ccaggcatct
     3361 tcaacagtac cacctcagat gctttcctgc agtctaaaat gaggaatatt ttttctggga
     3421 gattgacagc agttgaactg gttgacatac tggatcacta tactaagaat tgtgacatga
     3481 tggttggcaa ccaagcccta atccagggtt taaaaaaaga aaagtttgac ctgctactgg
     3541 ttgaccccaa tgatatgtgt ggatttgtga tcgcccatct tttaggcgtt aagtatgctg
     3601 tattttctac tggcctttgg tatcctgctg aagtaggagc acctgctcct ttagcttatg
     3661 tcccagagtt taactcactc ctcacagacc gcatgaactt cctggaaagg atgaaaaata
     3721 ctggtgttta cctcatatcc agaatagggg ttagctttct ggttctcccg aaatatgaga
     3781 ggataatgca gaagtacaac ctgctgcctg caaagtccat gtatgatttg gttcatgggt
     3841 ccagcctatg gatgttatgt actgacgtag cactggagtt tccaagacca acgctgccta
     3901 atgttgtgta tgttggagga atcctgacaa agccggccag cccactgcca gaagatctgc
     3961 agaggtgggt aagtggtgct caggaacatg gctttgtcct ggtgtctttt ggagctggtg
     4021 tcaagtatct gtcagaagac attgccaaca aactggcagg agctctgggg agattgcctc
     4081 aaaaagtgat ttggagattt tctggaacca aaccaaagaa cctaggaaac aacactaagc
     4141 tcatagaatg gctgcctcaa aatgacctgc ttggccattc aaatatcaga gccttcctaa
     4201 gccatggtgg tttgaacagt atatttgaaa ctatgtatca tggtgtccct gtagtgggaa
     4261 tcccactgtt tggagaccat tatgatacta tgactagagt acaggcaaaa ggcatgggga
     4321 tcttgttaga atggaataca gttactgaag gagagctgta tgatgccctg gtgaaagtta
     4381 tcaataatcc cagttatcgg cagagggctc agaagttatc ggaaattcac aaggatcaac
     4441 ccggccaccc tgtcaatcgg actacctatt ggatagacta cattcttcgc catgacggag
     4501 cccgtcacct ccgttctgct gtccatcaga tctctttctg ccagtatttc ctactggata
     4561 ttgcctttgt gcttctgctt ggtgctgtct tgctctactt catattgtcc tatgtgacaa
     4621 aatttatcta tagaaaaatc aaaagtctat ggtctaaaaa tgagcatagc acagttaatg
     4681 gacattacca aaatggaatc cgcaatggca aatacaaagg caatggtcgt gttaaacacg
     4741 aaaagaaggt gagaggatcc gattataagg atgatgacga caaaggagac tataaagacg
     4801 acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     4861 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     4921 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     4981 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     5041 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     5101 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     5161 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     5221 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     5281 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     5341 acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc
     5401 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     5461 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     5521 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5581 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5641 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5701 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5761 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5821 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5881 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5941 acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa
     6001 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     6061 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     6121 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     6181 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     6241 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     6301 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     6361 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     6421 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6481 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6541 cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc
     6601 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     6661 ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     6721 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6781 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6841 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6901 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     7081 ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     7141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     7201 tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg
     7261 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7321 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7381 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7441 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7501 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7561 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7621 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7681 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7741 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7801 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7861 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7921 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7981 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8041 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8101 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8161 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8221 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8281 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8341 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8401 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8461 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8521 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8581 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8641 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8701 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8761 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8821 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8881 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8941 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9001 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9061 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9121 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9181 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9241 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9301 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9361 aaaaataa
//
