LOCUS       V039537                 9523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039537
VERSION     V039537
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..4210
                     /label="HS3ST4(NM_006040)"
                     /note="HS3ST4(NM_006040)"
                     /gene="HS3ST4"
     CDS             4226..4291
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4311..4810
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4831..5430
                     /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    5444..6032
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6239..6872
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7001..7017)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7025..7041
                     /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(7049..7079)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7094..7115
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7403..7991)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8162..9022)
                     /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(9023..9127)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9175..9309
                     /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 ccatggcccg gtggcccgca cctcctccgc ctccgcctcc
     2881 gcctccacct ctggccgcgc cgccgccgcc cggcgcctct gctaaggggc cgccggcgcg
     2941 caagctgctt tttatgtgca ccttgtccct gtctgtcacc tacctgtgct acagcctcct
     3001 gggcggctcg ggctccctgc aattccctct ggcgctgcag gagtcgccgg gcgccgccgc
     3061 cgagcccccg ccgagcccgc cgccaccctc tctgctgcct acccccgtgc gcctcggcgc
     3121 cccctcgcag ccgcccgcgc cgccgccgct ggacaacgcg agccacgggg agccgcccga
     3181 gcccccagag cagccagccg cccccgggac cgacggctgg gggctgccga gcggcggcgg
     3241 aggcgcccag gacgcctggc tccggacccc gctggccccc agcgagatga tcacggctca
     3301 gagcgcgctg ccggagaggg aagcgcagga gtccagcacc accgacgagg atctcgcagg
     3361 ccggagagcg gccaacggga gcagcgagag gggcggcgcc gtcagcaccc ccgactatgg
     3421 ggagaagaag ctgccacagg cgctcatcat cggggtcaag aaaggaggga cccgcgcgct
     3481 gctggaggcg atccgcgtgc acccggacgt gcgggcggtg ggcgtagagc cgcacttctt
     3541 cgacaggaac tacgaaaagg ggttggagtg gtacagaaat gtgatgccca agactttgga
     3601 tgggcaaata accatggaga agactccaag ttactttgtg acaaatgagg ctcccaagcg
     3661 cattcactcc atggccaagg acatcaaact gattgtggtg gtgagaaacc ccgtgaccag
     3721 ggccatctct gactacacgc agacactgtc aaagaaaccc gagatcccca cctttgaggt
     3781 gctggccttc aaaaaccgga ccctcgggct gatcgatgct tcctggagtg ccattcgaat
     3841 agggatctat gcgctgcatc tggaaaactg gctccagtat ttccccctct cccagatcct
     3901 ctttgtcagt ggtgagcgac tcattgtgga ccccgccggg gaaatggcca aagtacagga
     3961 ttttctaggc ctcaaacgtg ttgtgactga gaagcatttc tatttcaaca aaaccaaggg
     4021 gttcccttgc ctaaagaagc cagaagacag cagtgccccg aggtgcttag gcaagagcaa
     4081 aggtcggact catcctcgca ttgacccaga tgtcatccac agactgagga aattctacaa
     4141 acccttcaac ttgatgtttt accaaatgac tggtcaagat tttcagtggg aacaggaaga
     4201 gggtgataaa ggtggaggcg gatccgacta caaagaccat gacggtgatt ataaagatca
     4261 tgacatcgac tacaaggatg acgatgacaa gtagtctaga taattctacc gggtagggga
     4321 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     4381 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     4441 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     4501 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     4561 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     4621 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     4681 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     4741 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     4801 gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc
     4861 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4921 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4981 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5041 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5101 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5161 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5221 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5281 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5341 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5401 tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta
     5461 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5521 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5581 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5641 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5701 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5761 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5821 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5881 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5941 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6001 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     6061 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     6121 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     6181 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     6241 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     6301 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
     6361 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
     6421 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     6481 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
     6541 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
     6601 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
     6661 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     6721 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6781 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6841 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6901 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg
     6961 ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7021 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7081 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7141 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8221 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8341 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8581 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8821 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9001 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9061 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9121 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg
     9181 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     9241 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     9301 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
     9361 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
     9421 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt
     9481 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
