LOCUS       V035048                 9502 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035048
VERSION     V035048
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..4258
                     /label="FOXA1(NM_004496)"
                     /note="FOXA1(NM_004496)"
                     /gene="FOXA1"
     promoter        4290..4789
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4810..5409
                     /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    5423..6011
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6218..6851
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6980..6996)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7004..7020
                     /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(7028..7058)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7073..7094
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7382..7970)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8141..9001)
                     /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(9002..9106)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9154..9288
                     /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 ccatgttagg aactgtgaag atggaagggc atgaaaccag
     2881 cgactggaac agctactacg cagacacgca ggaggcctac tcctccgtcc cggtcagcaa
     2941 catgaactca ggcctgggct ccatgaactc catgaacacc tacatgacca tgaacaccat
     3001 gactacgagc ggcaacatga ccccggcgtc cttcaacatg tcctatgcca acccgggcct
     3061 aggggccggc ctgagtcccg gcgcagtagc cggcatgccg gggggctcgg cgggcgccat
     3121 gaacagcatg actgcggccg gcgtgacggc catgggtacg gcgctgagcc cgagcggcat
     3181 gggcgccatg ggtgcgcagc aggcggcctc catgaatggc ctgggcccct acgcggccgc
     3241 catgaacccg tgcatgagcc ccatggcgta cgcgccgtcc aacctgggcc gcagccgcgc
     3301 gggcggcggc ggcgacgcca agacgttcaa gcgcagctac ccgcacgcca agccgcccta
     3361 ctcgtacatc tcgctcatca ccatggccat ccagcaggcg cccagcaaga tgctcacgct
     3421 gagcgagatc taccagtgga tcatggacct cttcccctat taccggcaga accagcagcg
     3481 ctggcagaac tccatccgcc actcgctgtc cttcaatgac tgcttcgtca aggtggcacg
     3541 ctccccggac aagccgggca agggctccta ctggacgctg cacccggact ccggcaacat
     3601 gttcgagaac ggctgctact tgcgccgcca gaagcgcttc aagtgcgaga agcagccggg
     3661 ggccggcggc gggggcggga gcggaagcgg gggcagcggc gccaagggcg gccctgagag
     3721 ccgcaaggac ccctctggcg cctctaaccc cagcgccgac tcgcccctcc atcggggtgt
     3781 gcacgggaag accggccagc tagagggcgc gccggccccc gggcccgccg ccagccccca
     3841 gactctggac cacagtgggg cgacggcgac agggggcgcc tcggagttga agactccagc
     3901 ctcctcaact gcgcccccca taagctccgg gcccggggcg ctggcctctg tgcccgcctc
     3961 tcacccggca cacggcttgg caccccacga gtcccagctg cacctgaaag gggaccccca
     4021 ctactccttc aaccacccgt tctccatcaa caacctcatg tcctcctcgg agcagcagca
     4081 taagctggac ttcaaggcat acgaacaggc actgcaatac tcgccttacg gctctacgtt
     4141 gcccgccagc ctgcctctag gcagcgcctc ggtgaccacc aggagcccca tcgagccctc
     4201 agccctggag ccggcgtact accaaggtgt gtattccaga cccgtcctaa acacttccta
     4261 gggatcccgc gactctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     4321 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     4381 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     4441 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     4501 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     4561 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     4621 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     4681 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     4741 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
     4801 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4861 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4921 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4981 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5041 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5101 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5161 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5221 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5281 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5341 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5401 ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     5461 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5521 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5581 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5641 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5701 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5761 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5821 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5881 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5941 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6001 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     6061 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     6121 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     6181 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa
     6241 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag
     6301 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag
     6361 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg
     6421 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg
     6481 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag
     6541 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt
     6601 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt
     6661 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6721 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6781 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6841 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     6901 atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct
     6961 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7021 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7081 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7141 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7201 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7261 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7321 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7561 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7801 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8221 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8341 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8581 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8641 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8821 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9001 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9061 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9121 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt
     9181 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9241 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     9301 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     9361 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     9421 tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa
     9481 tatgtactac aaacttagta gt
//
