LOCUS       V025599                 9765 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025599
VERSION     V025599
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..4521
                     /label="ATP5F1A(NM_004046)"
                     /note="ATP5F1A(NM_004046)"
                     /gene="ATP5F1A"
     promoter        4553..5052
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5073..5672
                     /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    5686..6274
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6481..7114
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7243..7259)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7267..7283
                     /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(7291..7321)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7336..7357
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7645..8233)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8404..9264)
                     /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(9265..9369)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9417..9551
                     /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 tcatgctgtc cgtgcgcgtt
     2881 gctgcggccg tggtccgcgc ccttcctcgg cgggccggac tggtctccag aaatgctttg
     2941 ggttcatctt tcattgctgc aaggaacttc catgcctcta acactcatct tcaaaagact
     3001 gggactgctg agatgtcctc tattcttgaa gagcgtattc ttggagctga tacctctgtt
     3061 gatcttgaag aaactgggcg tgtcttaagt attggtgatg gtattgcccg cgtacatggg
     3121 ctgaggaatg ttcaagcaga agaaatggta gagttttctt caggcttaaa gggtatgtcc
     3181 ttgaacttgg aacctgacaa tgttggtgtt gtcgtgtttg gaaatgataa actaattaag
     3241 gaaggagata tagtgaagag gacaggagcc attgtggacg ttccagttgg tgaggagctg
     3301 ttgggtcgtg tagttgatgc ccttggtaat gctattgatg gaaagggtcc aattggttcc
     3361 aagacgcgta ggcgagttgg tctgaaagcc cccggtatca ttcctcgaat ttcagtgcgg
     3421 gaaccaatgc agactggcat taaggctgtg gatagcttgg tgccaattgg tcgtggtcag
     3481 cgtgaactga ttattggtga ccgacagact gggaaaacct caattgctat tgacacaatc
     3541 attaaccaga aacgtttcaa tgatggatct gatgaaaaga agaagctgta ctgtatttat
     3601 gttgctattg gtcaaaagag atccactgtt gcccagttgg tgaagagact tacagatgca
     3661 gatgccatga agtacaccat tgtggtgtcg gctacggcct cggatgctgc cccacttcag
     3721 tacctggctc cttactctgg ctgttccatg ggagagtatt ttagagacaa tggcaaacat
     3781 gctttgatca tctatgacga cttatccaaa caggctgttg cttaccgtca gatgtctctg
     3841 ttgctccgcc gaccccctgg tcgtgaggcc tatcctggtg atgtgttcta cctacactcc
     3901 cggttgctgg agagagcagc caaaatgaac gatgcttttg gtggtggctc cttgactgct
     3961 ttgccagtca tagaaacaca ggctggtgat gtgtctgctt acattccaac aaatgtcatt
     4021 tccatcactg acggacagat cttcttggaa acagaattgt tctacaaagg tatccgccct
     4081 gcaattaacg ttggtctgtc tgtatctcgt gtcggatccg ctgcccaaac cagggctatg
     4141 aagcaggtag caggtaccat gaagctggaa ttggctcagt atcgtgaggt tgctgctttt
     4201 gcccagttcg gttctgacct cgatgctgcc actcaacaac ttttgagtcg tggcgtgcgt
     4261 ctaactgagt tgctgaagca aggacagtat tctcccatgg ctattgaaga acaagtggct
     4321 gttatctatg cgggtgtaag gggatatctt gataaactgg agcccagcaa gattacaaag
     4381 tttgagaatg ctttcttgtc tcatgtcgtc agccagcacc aagccttgtt gggcactatc
     4441 agggctgatg gaaagatctc agaacaatca gatgcaaagc tgaaagagat tgtaacaaat
     4501 ttcttggctg gatttgaagc ttaaggatcc cgcgactcta gataattcta ccgggtaggg
     4561 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     4621 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     4681 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     4741 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     4801 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     4861 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     4921 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     4981 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     5041 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     5101 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     5161 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     5221 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5281 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5341 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5401 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5461 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5521 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5581 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5641 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat
     5701 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5761 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5821 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5881 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5941 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6001 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     6061 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     6121 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6181 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6241 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc
     6301 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     6361 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     6421 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac
     6481 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     6541 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
     6601 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
     6661 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     6721 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
     6781 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
     6841 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
     6901 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
     6961 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7021 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7081 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     7141 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag
     7201 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     7261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     8041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     8101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8281 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8341 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8401 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8461 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8521 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8581 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8641 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8701 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8761 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8821 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8881 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8941 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     9001 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     9061 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     9121 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9181 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9241 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9301 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9361 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact
     9421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     9481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     9541 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat
     9601 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg
     9661 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt
     9721 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
