LOCUS       V046353                 8695 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046353
VERSION     V046353
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3549
                     /label="CSNK1E(NM_152221)"
                     /note="CSNK1E(NM_152221)"
                     /gene="CSNK1E"
     CDS             3556..3621
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3665..3876
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3889..4157
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4184..4783
                     /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    4790..5378
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5452..5685
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5757..5878
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5897..6032
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6070..6086)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6094..6110
                     /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(6118..6148)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6163..6184
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6472..7060)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7231..8091)
                     /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"
     promoter        complement(8092..8196)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8670..8686
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggagcta cgtgtgggga acaagtaccg cctgggacgg
     2341 aagatcggga gcgggtcctt cggagatatc tacctgggtg ccaacatcgc ctctggtgag
     2401 gaagtcgcca tcaagctgga gtgtgtgaag acaaagcacc cccagctgca catcgagagc
     2461 aagttctaca agatgatgca gggtggcgtg gggatcccgt ccatcaagtg gtgcggagct
     2521 gagggcgact acaacgtgat ggtcatggag ctgctggggc ctagcctcga ggacctgttc
     2581 aacttctgtt cccgcaaatt cagcctcaag acggtgctgc tcttggccga ccagatgatc
     2641 agccgcatcg agtatatcca ctccaagaac ttcatccacc gggacgtcaa gcccgacaac
     2701 ttcctcatgg ggctggggaa gaagggcaac ctggtctaca tcatcgactt cggcctggcc
     2761 aagaagtacc gggacgcccg cacccaccag cacattccct accgggaaaa caagaacctg
     2821 accggcacgg cccgctacgc ttccatcaac acgcacctgg gcattgagca aagccgtcga
     2881 gatgacctgg agagcctggg ctacgtgctc atgtacttca acctgggctc cctgccctgg
     2941 caggggctca aagcagccac caagcgccag aagtatgaac ggatcagcga gaagaagatg
     3001 tcaacgccca tcgaggtcct ctgcaaaggc tatccctccg aattctcaac atacctcaac
     3061 ttctgccgct ccctgcggtt tgacgacaag cccgactact cttacctacg tcagctcttc
     3121 cgcaacctct tccaccggca gggcttctcc tatgactacg tctttgactg gaacatgctg
     3181 aaattcggtg cagcccggaa tcccgaggat gtggaccggg agcggcgaga acacgaacgc
     3241 gaggagagga tggggcagct acgggggtcc gcgacccgag ccctgccccc tggcccaccc
     3301 acgggggcca ctgccaaccg gctccgcagt gccgccgagc ccgtggcttc cacgccagcc
     3361 tcccgcatcc agccggctgg caatacttct cccagagcga tctcgcgggt cgaccgggag
     3421 aggaaggtga gtatgaggct gcacaggggt gcgcccgcca acgtctcctc ctcagacctc
     3481 actgggcggc aagaggtctc ccggatccca gcctcacaga caagtgtgcc atttgaccat
     3541 ctcgggaagg gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3601 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3661 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3721 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3781 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3841 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     3901 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3961 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     4021 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     4081 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     4141 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     4201 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     4261 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     4321 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4381 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4441 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4501 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4561 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4621 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4681 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4741 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     4801 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4861 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4921 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4981 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5041 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5101 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5161 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5221 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5281 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5341 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     5401 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5461 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     5521 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5581 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5641 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     5701 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     5761 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5821 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5881 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5941 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6001 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     6061 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6121 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6181 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6901 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8221 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8281 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     8341 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8401 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     8461 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     8521 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     8581 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     8641 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
