LOCUS       V041429                10009 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041429
VERSION     V041429
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2843..4705
                     /label="Papss2(NM_011864)"
                     /note="Papss2(NM_011864)"
                     /gene="Papss2"
     CDS             4712..4777
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4797..5296
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5317..5916
                     /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    5930..6518
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6725..7358
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7487..7503)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7511..7527
                     /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(7535..7565)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7580..7601
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7889..8477)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8648..9508)
                     /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(9509..9613)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9661..9795
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattcgcca ccatgtctgc aaatttcaaa atgaaccata aaagagacca
     2881 gcaaaaatcc accaatgtgg tctaccaggc ccatcatgtg agcaggaaca agagaggaca
     2941 agtggttgga accaggggag gattccgagg atgtaccgtg tggctaacag gtctctctgg
     3001 tgctgggaaa acaaccataa gctttgcttt ggaagagtac cttgtatctc acgccatccc
     3061 atgttactct ctggatgggg acaatgtccg tcatggcctt aataagaacc tgggattctc
     3121 tgcgggggac cgagaagaga atatccgccg gatcgcggag gtggccaggc tctttgccga
     3181 cgccggcctg gtttgcatca ccagctttat ctctcctttt gcaaaggatc gtgagaatgc
     3241 ccgaaaaatc cacgaatcag caggactccc gttctttgag atctttgtag atgcgccttt
     3301 aaatatctgt gaaagccgag acgtaaaagg actctacaaa cgagcccgag caggagagat
     3361 taaagggttt acaggcatcg attctgacta tgagaaacct gaaactccag agtgtgtgct
     3421 gaagaccaac ctgtcttcag taagcgactg tgtgcaacag gtggtggaac ttttgcagga
     3481 gcagaacatt gtaccccaca ccaccatcaa aggcatccac gaactctttg tgccagaaaa
     3541 caaagtcgat caaatccgag ctgaggcaga gactctccca tcactaccaa ttaccaagct
     3601 ggatctgcag tgggtgcaga ttctgagtga aggctgggcc actcccctca aaggctttat
     3661 gcgggagaag gagtacttgc aaactctaca cttcgacact ctactggacg gcgtggttcc
     3721 ccgtgatgga gtcatcaaca tgagtattcc cattgtattg cccgtttctg cggatgacaa
     3781 ggcacggctc gaagggtgca gcaaatttgc cttgatgtac gaaggtcgga gggtcgctct
     3841 attacaggac cctgaattct atgagcatag gaaagaggag cgctgttctc gtgtgtgggg
     3901 aacagccact gcaaagcacc cccatatcaa aatggtgatg gaaagtgggg actggcttgt
     3961 tggtggagac ctacaggtgc tagagagaat aaggtgggac gatgggctgg accaataccg
     4021 ccttacgcct ctggagctca aacagaagtg taaagacatg aatgctgatg ccgtgtttgc
     4081 attccagttg cgcaatcctg tccacaatgg tcatgccctc ctgatgcagg acacccgccg
     4141 caggctcctg gagaggggtt acaagcaccc agtcctcctg ctccaccctc ttgggggctg
     4201 gaccaaggac gatgacgtac ctctggaatg gaggatgaaa cagcatgcag ctgtactgga
     4261 ggaaagggtc ctggatccca agtcaactat tgttgccatc tttccatctc ctatgttata
     4321 tgctggtccc acagaggtcc agtggcattg cagatgccgg atgattgcag gagccaattt
     4381 ctacattgtg ggtagggatc ccgcaggaat gccccatcct gagacaaaga aagacctata
     4441 tgaacccacc cacgggggca aggtcttgag tatggcccct ggccttacct ctgtggaaat
     4501 aattccgttc cgagtggctg cctacaataa aattaaaaag gccatggact tttatgatcc
     4561 agcaaggcac gaggagtttg acttcatctc aggaactcgc atgaggaagc tcgcccggga
     4621 aggagaagat cccccagatg gcttcatggc cccgaaagcg tggaaagtgt tgacagatta
     4681 ctacaggtct ctggagaaga ccaacggatc cgactacaaa gaccatgacg gtgattataa
     4741 agatcatgac atcgactaca aggatgacga tgacaagtag tctagataat tctaccgggt
     4801 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     4861 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     4921 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     4981 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     5041 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     5101 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     5161 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     5221 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     5281 atctccgggc ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg
     5341 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     5401 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     5461 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5521 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5581 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5641 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5701 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5761 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5821 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5881 cgcacctggt gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct
     5941 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     6001 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     6061 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     6121 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     6181 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     6241 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     6301 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     6361 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     6421 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6481 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga
     6541 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     6601 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     6661 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg
     6721 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     6781 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat
     6841 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag
     6901 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac
     6961 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc
     7021 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt
     7081 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc
     7141 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat
     7201 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7261 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7321 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     7381 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg
     7441 ctagcgtttt accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     7501 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7561 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7621 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7681 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7741 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7801 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7861 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7921 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7981 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     8041 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     8101 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     8161 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     8221 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     8281 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     8341 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8401 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     8461 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8521 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8581 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8641 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8701 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8761 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8821 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8881 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8941 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     9001 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     9061 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     9121 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     9181 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     9241 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     9301 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     9361 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     9421 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9481 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     9541 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9601 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     9661 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     9721 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     9781 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     9841 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     9901 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     9961 gtttttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
