LOCUS       V047035                10363 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047035
VERSION     V047035
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..5119
                     /label="Cul4a(NM_146207)"
                     /note="Cul4a(NM_146207)"
                     /gene="Cul4a"
     promoter        5151..5650
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5671..6270
                     /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    6284..6872
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7079..7712
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7841..7857)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7865..7881
                     /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(7889..7919)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7934..7955
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8243..8831)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9002..9862)
                     /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(9863..9967)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10015..10149
                     /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 ccatggcgga cgagggccct cggaagggca gcgtctcggc
     2881 tctcatgggc cgcaccaacg gcctcaccaa gcccgcggcg ttggccggtg gccccgccaa
     2941 gccggggggc acgggaggct ccaggaagct ggtcatcaag aacttccgag acaggccgcg
     3001 gctgcctgac aactacactc aggacacgtg gcggaagctt cacgaagcgg tcaaggccat
     3061 ccagagcagc acatccatca ggtacaacct ggaggagctg taccaggctg ttgaaaatct
     3121 ttgttctcac aaagtctccc caacgctcta caagcagctg cgccaggtct gtgaagacca
     3181 tgtccaggcc cagatcctcc cattcagaga agactcacta gacagcgttt tatttctaaa
     3241 gaagattaac acatgctggc aagatcactg cagacaaatg atcatgatca gaagcatctt
     3301 cctgtttctg gatcgaacct acgtcctcca gaattccatg ctgccttcta tctgggacat
     3361 gggcctagag ctgtttagga accacatcat cagcgacagg atggtgcaga gtaagaccat
     3421 tgatgggatc ctgctgctga ttgggcgtga acgaagtgga gaggctgtgg acaggagtct
     3481 gcttcggagt ttgctaagca tgctctcaga tctccaggtg tacaaagact catttgagct
     3541 gaagttcttg gaagagacta actgtctata cgctgctgaa ggccagaggc tgatgcagga
     3601 cagggaggtt ccagaatacc ttaaccatgt gagcaaacgt ctagaagagg aagcagaccg
     3661 agtgatcacc tacttagacc acagcacaca aaaaccgttg attgcctgtg tggagaaaca
     3721 gctgttagga gaacatttga cagcaattct acagaaaggc ctagagcacc tgctggatga
     3781 gaacagggtg cccgacctca cccagatgta ccagctcttc agccgggtga aaggagggca
     3841 gcatgcgctg ctgcagcact ggagcgagta catcaagacc tttggaacaa ccattgtcat
     3901 caatcctgag aaagacaaag acatggtcca agacctgctg gactttaagg acaaagtgga
     3961 ccacgtggtg gaggtgtgct tccagaggaa cgagcgcttc atcaacctga tgaaggagtc
     4021 cttcgagacg ttcatcaaca agagaccaaa caagcctgca gagctcattg ccaagcacgt
     4081 ggactcaaag ttacgggccg gcaacaaaga agccacagat gaggaactgg agaggatcct
     4141 ggacaagatc atgattctct tccgtttcat ccatggtaaa gatgtctttg aagcgtttta
     4201 taagaaagac ttagcaaaaa ggctgctggt ggggaagagc gcgtcggtgg acgcggagaa
     4261 gtccatgctg tcgaagctca agcacgagtg cggagctgcg tttaccagca aactggaggg
     4321 catgttcaag gacatggagc tctccaaaga catcatggtt cacttcaagc agcatatgca
     4381 gaaccagagt gcgccaggcc ccattgacct cacggtgaac atactcacca tgggctactg
     4441 gcccacgtac acacccatgg aagtgcacct ccctccagag atggtcagac ttcaggaagt
     4501 attcaagacg ttctacctgg gcaagcacag tgggcggaag ctgcaatggc agacaacgct
     4561 gggacacgcc gtgctgaagg cggactttaa ggaggggaag aaggagttcc aggtatccct
     4621 cttccagacg ctggtgctgc tcatgttcaa cgaaggggat ggatttagct ttgaagaaat
     4681 aaaaatggcc actggcatag aggacagtga gttgcgaaga acactacagt ctctggcctg
     4741 tggtaaagca cgggtcctaa ttaaaagtcc caaaggaaag gaggtggaag atggagataa
     4801 gttcatcttc aatgcagact tcaaacacaa gttgttcaga ataaagatca atcaaatcca
     4861 gatgaaggag accgttgaag aacaggttag cactacagag agagtattcc aggatagaca
     4921 gtaccagatc gacgctgcta tagtcagaat aatgaaaatg aggaagacac ttggtcataa
     4981 tcttctagtt tctgaattat acaatcagct gaaatttcca gtgaagcctg gagatttgaa
     5041 aaaaaggatt gagtcactta tagacagaga ctatatggaa cgagacaaag acagtccaaa
     5101 tcagtaccac tacgtggcat gaggatcccg cgactctaga taattctacc gggtagggga
     5161 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     5221 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     5281 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     5341 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     5401 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     5461 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     5521 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     5581 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     5641 gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc
     5701 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5761 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5821 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5881 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5941 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     6001 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     6061 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     6121 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     6181 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     6241 tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta
     6301 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6361 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6421 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6481 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6541 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6601 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6661 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     6721 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6781 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6841 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     6901 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     6961 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     7021 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     7081 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     7141 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
     7201 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
     7261 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     7321 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
     7381 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
     7441 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
     7501 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     7561 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7621 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7681 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     7741 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg
     7801 ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7861 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7921 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     8041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     8101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     8161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9961 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg
    10021 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
    10081 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    10141 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
    10201 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
    10261 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt
    10321 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
