LOCUS       V033205                10564 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033205
VERSION     V033205
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..5320
                     /label="HIF1A(NM_001530)"
                     /note="HIF1A(NM_001530)"
                     /gene="HIF1A"
     promoter        5352..5851
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5872..6471
                     /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    6485..7073
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7280..7913
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(8042..8058)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8066..8082
                     /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(8090..8120)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8135..8156
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8444..9032)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9203..10063)
                     /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(10064..10168)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10216..10350
                     /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 ccatggaggg cgccggcggc gcgaacgaca agaaaaagat
     2881 aagttctgaa cgtcgaaaag aaaagtctcg agatgcagcc agatctcggc gaagtaaaga
     2941 atctgaagtt ttttatgagc ttgctcatca gttgccactt ccacataatg tgagttcgca
     3001 tcttgataag gcctctgtga tgaggcttac catcagctat ttgcgtgtga ggaaacttct
     3061 ggatgctggt gatttggata ttgaagatga catgaaagca cagatgaatt gcttttattt
     3121 gaaagccttg gatggttttg ttatggttct cacagatgat ggtgacatga tttacatttc
     3181 tgataatgtg aacaaataca tgggattaac tcagtttgaa ctaactggac acagtgtgtt
     3241 tgattttact catccatgtg accatgagga aatgagagaa atgcttacac acagaaatgg
     3301 ccttgtgaaa aagggtaaag aacaaaacac acagcgaagc ttttttctca gaatgaagtg
     3361 taccctaact agccgaggaa gaactatgaa cataaagtct gcaacatgga aggtattgca
     3421 ctgcacaggc cacattcacg tatatgatac caacagtaac caacctcagt gtgggtataa
     3481 gaaaccacct atgacctgct tggtgctgat ttgtgaaccc attcctcacc catcaaatat
     3541 tgaaattcct ttagatagca agactttcct cagtcgacac agcctggata tgaaattttc
     3601 ttattgtgat gaaagaatta ccgaattgat gggatatgag ccagaagaac ttttaggccg
     3661 ctcaatttat gaatattatc atgctttgga ctctgatcat ctgaccaaaa ctcatcatga
     3721 tatgtttact aaaggacaag tcaccacagg acagtacagg atgcttgcca aaagaggtgg
     3781 atatgtctgg gttgaaactc aagcaactgt catatataac accaagaatt ctcaaccaca
     3841 gtgcattgta tgtgtgaatt acgttgtgag tggtattatt cagcacgact tgattttctc
     3901 ccttcaacaa acagaatgtg tccttaaacc ggttgaatct tcagatatga aaatgactca
     3961 gctattcacc aaagttgaat cagaagatac aagtagcctc tttgacaaac ttaagaagga
     4021 acctgatgct ttaactttgc tggccccagc cgctggagac acaatcatat ctttagattt
     4081 tggcagcaac gacacagaaa ctgatgacca gcaacttgag gaagtaccat tatataatga
     4141 tgtaatgctc ccctcaccca acgaaaaatt acagaatata aatttggcaa tgtctccatt
     4201 acccaccgct gaaacgccaa agccacttcg aagtagtgct gaccctgcac tcaatcaaga
     4261 agttgcatta aaattagaac caaatccaga gtcactggaa ctttctttta ccatgcccca
     4321 gattcaggat cagacaccta gtccttccga tggaagcact agacaaagtt cacctgagcc
     4381 taatagtccc agtgaatatt gtttttatgt ggatagtgat atggtcaatg aattcaagtt
     4441 ggaattggta gaaaaacttt ttgctgaaga cacagaagca aagaacccat tttctactca
     4501 ggacacagat ttagacttgg agatgttagc tccctatatc ccaatggatg atgacttcca
     4561 gttacgttcc ttcgatcagt tgtcaccatt agaaagcagt tccgcaagcc ctgaaagcgc
     4621 aagtcctcaa agcacagtta cagtattcca gcagactcaa atacaagaac ctactgctaa
     4681 tgccaccact accactgcca ccactgatga attaaaaaca gtgacaaaag accgtatgga
     4741 agacattaaa atattgattg catctccatc tcctacccac atacataaag aaactactag
     4801 tgccacatca tcaccatata gagatactca aagtcggaca gcctcaccaa acagagcagg
     4861 aaaaggagtc atagaacaga cagaaaaatc tcatccaaga agccctaacg tgttatctgt
     4921 cgctttgagt caaagaacta cagttcctga ggaagaacta aatccaaaga tactagcttt
     4981 gcagaatgct cagagaaagc gaaaaatgga acatgatggt tcactttttc aagcagtagg
     5041 aattggaaca ttattacagc agccagacga tcatgcagct actacatcac tttcttggaa
     5101 acgtgtaaaa ggatgcaaat ctagtgaaca gaatggaatg gagcaaaaga caattatttt
     5161 aataccctct gatttagcat gtagactgct ggggcaatca atggatgaaa gtggattacc
     5221 acagctgacc agttatgatt gtgaagttaa tgctcctata caaggcagca gaaacctact
     5281 gcagggtgaa gaattactca gagctttgga tcaagttaac tgaggatccc gcgactctag
     5341 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     5401 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     5461 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     5521 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     5581 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     5641 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     5701 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc
     5761 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     5821 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag
     5881 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     5941 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     6001 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     6061 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     6121 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     6181 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6241 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6301 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6361 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6421 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg
     6481 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     6541 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     6601 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     6661 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     6721 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6781 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6841 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     6901 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6961 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     7021 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     7081 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     7141 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     7201 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     7261 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt
     7321 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg
     7381 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
     7441 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
     7501 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
     7561 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct
     7621 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg
     7681 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct
     7741 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     7801 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     7861 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     7921 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     7981 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat
     8041 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     8101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     8161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     8221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     8281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8881 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     9001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     9061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     9121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    10021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    10081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    10141 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
    10201 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    10261 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    10321 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
    10381 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
    10441 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
    10501 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag
    10561 tagt
//
