LOCUS       V019681                 9243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019681
VERSION     V019681
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             2827..3999
                     /label="NAP1L1(NM_004537)"
                     /note="NAP1L1(NM_004537)"
                     /gene="NAP1L1"
     promoter        4031..4530
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4551..5150
                     /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    5164..5752
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5959..6592
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6721..6737)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6745..6761
                     /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(6769..6799)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6814..6835
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7123..7711)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7882..8742)
                     /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(8743..8847)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8895..9029
                     /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 gccaccatgg cagacattga caacaaagaa cagtctgaac ttgatcaaga tttggatgat
     2881 gttgaagaag tagaagaaga ggaaactggt gaagaaacaa aactcaaagc acgtcagcta
     2941 actgttcaga tgatgcaaaa tcctcagatt cttgcagccc ttcaagaaag acttgatggt
     3001 ctggtagaaa caccaacagg atacattgaa agcctgccta gggtagttaa aagacgagtg
     3061 aatgctctca aaaacctgca agttaaatgt gcacagatag aagccaaatt ctatgaggaa
     3121 gttcacgatc ttgaaaggaa gtatgctgtt ctctatcagc ctctatttga taagcgattt
     3181 gaaattatta atgcaattta tgaacctacg gaagaagaat gtgaatggaa accagatgaa
     3241 gaagatgaga tttcggagga attgaaagaa aaggccaaga ttgaagatga gaaaaaagat
     3301 gaagaaaaag aagaccccaa aggaattcct gaattttggt taactgtttt taagaatgtt
     3361 gacttgctca gtgatatggt tcaggaacac gatgaaccta ttctgaagca cttgaaagat
     3421 attaaagtga agttctcaga tgctggccag cctatgagtt ttgtcttaga atttcacttt
     3481 gaacccaatg aatattttac aaatgaagtg ctgacaaaga catacaggat gaggtcagaa
     3541 ccagatgatt ctgatccctt ttcttttgat ggaccagaaa ttatgggttg tacagggtgc
     3601 cagatagatt ggaaaaaagg aaagaatgtc actttgaaaa ctattaagaa gaagcagaaa
     3661 cacaagggac gtgggacagt tcgtactgtg actaaaacag tttccaatga ctctttcttt
     3721 aacttttttg cccctcctga agttcctgag agtggagatc tggatgatga tgctgaagct
     3781 atccttgctg cagacttcga aattggtcac tttttacgtg agcgtataat cccaagatca
     3841 gtgttatatt ttactggaga agctattgaa gatgatgatg atgattatga tgaagaaggt
     3901 gaagaagcgg atgaggaagg ggaagaagaa ggagatgagg aaaatgatcc agactatgac
     3961 ccaaagaagg atcaaaaccc agcagagtgc aagcagcagt gaggatcccg cgactctaga
     4021 taattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     4081 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     4141 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     4201 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     4261 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     4321 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     4381 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     4441 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     4501 ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt
     4561 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4621 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4681 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4741 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4801 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4861 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4921 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4981 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     5041 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5101 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg
     5161 aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5221 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5281 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5341 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5401 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5461 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5521 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     5581 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5641 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5701 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     5761 aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     5821 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     5881 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5941 taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     6001 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc
     6061 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag
     6121 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc
     6181 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat
     6241 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt
     6301 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
     6361 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc
     6421 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     6481 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     6541 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     6601 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     6661 aggccttgac attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg
     6721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7561 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7801 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8041 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8401 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8761 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8821 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8881 cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     8941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     9001 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
     9061 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
     9121 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
     9181 ctacaaactt agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     9241 agt
//
