LOCUS       V032603                 9874 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032603
VERSION     V032603
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..4570
                     /label="Nt5e(NM_011851)"
                     /note="Nt5e(NM_011851)"
                     /gene="Nt5e"
     CDS             4577..4642
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4662..5161
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5182..5781
                     /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    5795..6383
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6590..7223
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7352..7368)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7376..7392
                     /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(7400..7430)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7445..7466
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7754..8342)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8513..9373)
                     /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(9374..9478)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9526..9660
                     /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 ccatgcgtcc cgcggccgct aaggtaccca agtggctgct
     2881 tctcgcactg agcgctctac taccacagtg gcccgcagcc agtgcctggg agctcacgat
     2941 cctgcacaca aacgacgtgc acagccggct agagcagacc agcgatgact ccaccaagtg
     3001 cctcaacgcc agcctgtgtg tgggcggcgt ggcccggctc tttaccaagg tgcagcagat
     3061 ccgcaaggaa gaacccaacg tgctgttttt ggatgccgga gaccagtacc agggcaccat
     3121 ctggttcacc gtttacaaag gccttgaagt ggcacacttc atgaacatcc tgggctacga
     3181 tgctatggca ctgggaaatc atgaatttga taacggtgtg gaaggactga ttgatcccct
     3241 cctcagaaac gttaaatttc caattctgag cgcaaacatt aaggcacggg ggcctctagc
     3301 acatcagata tctggacttt ttctgccatc taaagttctc tctgttggcg gtgaggttgt
     3361 ggggattgtt ggatatactt caaaggaaac ccctttcctc tcaaatccag ggacaaattt
     3421 agtctttgaa gatgaaatct ctgcattgca gcctgaagta gataaactaa agacgctaaa
     3481 tgtgaataag atcatcgccc tggggcactc tggttttgag atggacaaac ttatcgctca
     3541 gaaagttcga ggtgtggaca tcgtggtggg aggacactcc aacacctttc tctacacagg
     3601 aaatccacct tccaaagaag tgcctgcggg gaagtaccca ttcatagtca ccgcagatga
     3661 tggacggcag gtgcctgtgg tccaggccta tgcctttggc aaatacctgg gctacctgaa
     3721 ggttgagttt gatgataaag gcaatgttat cacttcctat ggaaatccca ttcttctcaa
     3781 cagcagcatt cctgaagatg cgaccatcaa agcagacatt aaccaatgga ggataaaatt
     3841 agataattat tctacccagg aactcgggag aacgatcgtc tacctggatg gctccactca
     3901 gacgtgccgc ttcagggaat gcaacatggg aaacctgatc tgtgatgcca tgattaacaa
     3961 caacctcaga cacccagatg aaatgttttg gaaccacgtg tccatgtgca ttgtaaacgg
     4021 aggtggcatc cggtccccca ttgatgagaa gaacaatggt accatcacct gggagaacct
     4081 ggctgctgtg ctgccctttg gagggacatt tgacctcgtc caattaaaag ggtccaccct
     4141 gaagaaggct tttgagcaca gcgtgcatcg ctatggccag tccaccggag agttcctgca
     4201 agtgggtgga atccatgtgg tgtacgatat taaccgaaag ccctggaaca gagtggtcca
     4261 attagaagtt ctctgcacca agtgtcgagt gcccatctat gagcctcttg aaatggataa
     4321 agtgtataaa gtgaccctcc caagctatct ggccaacggt ggagatggat tccagatgat
     4381 aaaagatgaa ttactaaagc atgactctgg tgatcaagat atcagcgtgg tttctgaata
     4441 catctcaaaa atgaaagtag tttacccagc cgttgaaggg cggatcaagt tctctgcagc
     4501 aagtcattac cagggaagct ttcctttagt aattctttca ttttgggcaa tgatccttat
     4561 tttgtaccaa ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga
     4621 ctacaaggat gacgatgaca agtagtctag ataattctac cgggtagggg aggcgctttt
     4681 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     4741 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     4801 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     4861 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     4921 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     4981 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     5041 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     5101 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     5161 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     5221 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5281 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5341 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5401 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5461 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5521 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5581 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5641 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5701 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5761 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg
     5821 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5881 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5941 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6001 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6061 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6121 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6181 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     6241 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6301 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6361 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     6421 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6481 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6541 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     6601 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6661 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     6721 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     6781 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     6841 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     6901 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
     6961 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     7021 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7081 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7141 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7201 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     7261 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt
     7321 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9481 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca
     9541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     9661 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
     9721 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
     9781 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt
     9841 gtatttgtta aatatgtact acaaacttag tagt
//
