LOCUS       V025696                 9693 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025696
VERSION     V025696
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             2830..2856
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2863..4449
                     /label="ATP5F1B(NM_001686)"
                     /note="ATP5F1B(NM_001686)"
                     /gene="ATP5F1B"
     promoter        4481..4980
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5001..5600
                     /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    5614..6202
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6409..7042
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7171..7187)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7195..7211
                     /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(7219..7249)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7264..7285
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7573..8161)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8332..9192)
                     /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(9193..9297)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9345..9479
                     /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 gccaccatgt acccatacga cgtaccagat tacgctgaat tcatgttggg gtttgtgggt
     2881 cgggtggccg ctgctccggc ctccggggcc ttgcggagac tcaccccttc agcgtcgctg
     2941 cccccagctc agctcttact gcgggccgct ccgacggcgg tccatcctgt cagggactat
     3001 gcggcgcaaa catctccttc gccaaaagca ggcgccgcca ccgggcgcat cgtggcggtc
     3061 attggcgcag tggtggacgt ccagtttgat gagggactac caccaattct aaatgccctg
     3121 gaagtgcaag gcagggagac cagactggtt ttggaggtgg cccagcattt gggtgagagc
     3181 acagtaagga ctattgctat ggatggtaca gaaggcttgg ttagaggcca gaaagtactg
     3241 gattctggtg caccaatcaa aattcctgtt ggtcctgaga ctttgggcag aatcatgaat
     3301 gtcattggag aacctattga tgaaagaggt cccatcaaaa ccaaacaatt tgctcccatt
     3361 catgctgagg ctccagagtt catggaaatg agtgttgagc aggaaattct ggtgactggt
     3421 atcaaggttg tcgatctgct agctccctat gccaagggtg gcaaaattgg gctttttggt
     3481 ggtgctggag ttggcaagac tgtactgatc atggagttaa tcaacaatgt cgccaaagcc
     3541 catggtggtt actctgtgtt tgctggtgtt ggtgagagga cccgtgaagg caatgattta
     3601 taccatgaaa tgattgaatc tggtgttatc aacttaaaag atgccacctc taaggtagcg
     3661 ctggtatatg gtcaaatgaa tgaaccacct ggtgctcgtg cccgggtagc tctgactggg
     3721 ctgactgtgg ctgaatactt cagagaccaa gaaggtcaag atgtactgct atttattgat
     3781 aacatctttc gcttcaccca ggctggttca gaggtgtctg cattattggg ccgaatccct
     3841 tctgctgtgg gctatcagcc taccctggcc actgacatgg gtactatgca ggaaagaatt
     3901 accactacca agaagggatc tatcacctct gtacaggcta tctatgtgcc tgctgatgac
     3961 ttgactgacc ctgcccctgc tactacgttt gcccatttgg atgctaccac tgtactgtcg
     4021 cgtgccattg ctgagctggg catctatcca gctgtggatc ctctagactc cacctctcgt
     4081 atcatggatc ccaacattgt tggcagtgag cattacgatg ttgcccgtgg ggtgcaaaag
     4141 atcctgcagg actacaaatc cctccaggat atcattgcca tcctgggtat ggatgaactt
     4201 tctgaggaag acaagttgac cgtgtcccgt gcacggaaaa tacagcgttt cttgtctcag
     4261 ccattccagg ttgctgaggt cttcacaggt catatgggga agctggtacc cctgaaggag
     4321 accatcaaag gattccagca gattttggca ggtgaatatg accatctccc agaacaggcc
     4381 ttctatatgg tgggacccat tgaagaagct gtggcaaaag ctgataagct ggctgaagag
     4441 cattcatcgt gaggatcccg cgactctaga taattctacc gggtagggga ggcgcttttc
     4501 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     4561 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     4621 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     4681 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     4741 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     4801 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     4861 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     4921 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     4981 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     5041 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     5101 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     5161 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     5221 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     5281 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     5341 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5401 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5461 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5521 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5581 cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt
     5641 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5701 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5761 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5821 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5881 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5941 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6001 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     6061 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6121 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6181 tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg
     6241 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     6301 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     6361 acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa
     6421 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     6481 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
     6541 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
     6601 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
     6661 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
     6721 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
     6781 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
     6841 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6901 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6961 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7021 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     7081 cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc
     7141 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7201 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7261 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7321 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7381 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7441 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7741 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7981 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     8041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8221 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8281 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8341 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8401 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8461 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8521 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8581 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8641 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8701 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8761 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8821 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8881 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8941 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9001 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9061 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9121 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9181 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9301 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag
     9361 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     9421 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     9481 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
     9541 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
     9601 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg
     9661 tatttgttaa atatgtacta caaacttagt agt
//
