LOCUS       V018726                10335 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018726
VERSION     V018726
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             2827..5061
                     /label="PAPOLA(NM_032632)"
                     /note="PAPOLA(NM_032632)"
                     /gene="PAPOLA"
     CDS             5068..5091
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        5123..5622
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5643..6242
                     /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    6256..6844
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7051..7684
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7813..7829)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7837..7853
                     /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(7861..7891)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7906..7927
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8215..8803)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8974..9834)
                     /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(9835..9939)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9987..10121
                     /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 gccaccatgc cgtttccagt tacaacacag ggatcacaac aaacacaacc gccacagaag
     2881 cactatggca ttacttctcc tatcagctta gcagccccca aggagactga ctgcgtactt
     2941 acacagaaac taattgagac attgaaaccc tttggggttt ttgaagagga agaggaactg
     3001 cagcgcagga ttttaatttt gggaaaacta aataacctgg taaaagagtg gatacgagaa
     3061 atcagtgaaa gcaagaatct tccacaatct gtaattgaaa atgttggagg aaaaattttt
     3121 acatttggat cttacagatt aggagtgcat acaaaaggtg ctgatattga tgcgttgtgt
     3181 gttgcaccaa gacatgttga tcgaagtgac tttttcacct cattctatga taagttgaaa
     3241 ttacaggaag aagtaaaaga tttaagagct gttgaagagg cattcgtacc agttattaaa
     3301 ctctgttttg atgggataga gattgatatt ttgtttgcaa gattagcact gcagacaatt
     3361 cctgaagatt tggatctacg agatgacagt ctgctaaaaa atttagatat aagatgtata
     3421 agaagtctta acggttgcag ggtaaccgat gaaattttac atctagtacc aaacattgac
     3481 aacttcaggt taactctgag agctatcaaa ctatgggcca aacgccacaa catctattcc
     3541 aatatattag gtttcctcgg tggtgtttcc tgggctatgc tagtagcaag aacttgccag
     3601 ctttatccaa atgcaatagc atcaactctt gtacataaat ttttcttggt attttctaaa
     3661 tgggaatggc caaatccagt gctattgaaa cagcctgaag aatgcaatct taatttgcct
     3721 gtatgggacc caagggtaaa ccccagtgat aggtaccatc ttatgcctat aattacacca
     3781 gcatacccac aacagaactc cacgtacaat gtgtccgttt caacacggat ggtcatggtt
     3841 gaggagttta aacaaggtct tgctatcaca gatgaaattt tgctgagtaa ggcagagtgg
     3901 tccaaacttt ttgaagctcc aaacttcttt caaaagtaca agcattatat tgtacttcta
     3961 gcaagtgcac caacagaaaa acaacgcctg gaatgggtgg gcttggtgga atcaaaaatc
     4021 cgaatcctgg ttggaagctt ggagaagaat gaatttatta cactggctca tgtgaatccc
     4081 cagtcatttc cagcacccaa agaaaatccc gacaaggaag aatttcgcac gatgtgggtg
     4141 attgggttag tgtttaaaaa aacagaaaac tctgaaaacc tcagtgttga tctcacctat
     4201 gatattcagt ctttcacaga tacagtttat aggcaagcaa taaacagcaa gatgtttgag
     4261 gtggatatga aaattgctgc aatgcatgta aaaagaaagc aactccatca actactacct
     4321 aatcatgtgc ttcagaaaaa gaaaaagcat tcaacagaag gtgtcaaatt gacagctctc
     4381 aatgacagca gcctcgactt gtctatggac agtgataaca gcatgtctgt gccttcacct
     4441 actagtgcta cgaagaccag tccattgaac agttctggca gctctcaggg cagaaacagt
     4501 cctgctccag ctgtaacagc agcatctgtg accaacatac aggctactga agtttctgtg
     4561 ccacaagtaa attccagtga aagctcaggg ggtacatcga gtgaaagcat tcctcaaact
     4621 gccacacaac cagccatttc tccaccacca aagcctacgg tctccagagt tgtttcttca
     4681 acacgtctgg taaacccacc acctagatct tcaggaaatg cagcaacttc aggaaatgca
     4741 gcaacaaaaa tacctactcc tatagtagga gtcaagagga catcctcacc tcataaagaa
     4801 gagagtccca agaaaaccaa aacagaagag gatgaaacaa gtgaagatgc taactgtctt
     4861 gctttgagtg gacatgataa aacagaagca aaggaacaac ttgatacaga gacaagtaca
     4921 actcaatcag aaactattca gacagcggct tctctgttgg cctctcagaa aacatccagt
     4981 acagaccttt ctgatatccc tgctctccct gcaaatccta ttcctgttat caagaattca
     5041 ataaaactga gattgaatcg ggaattcgat tacaaggatg acgacgataa gtgaggatcc
     5101 cgcgactcta gataattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     5161 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     5221 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     5281 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     5341 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     5401 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     5461 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     5521 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     5581 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     5641 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     5701 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     5761 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5821 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5881 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5941 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     6001 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     6061 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     6121 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     6181 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     6241 gaccgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6301 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6361 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6421 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6481 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6541 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6601 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     6661 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6721 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6781 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6841 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     6901 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     6961 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     7021 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
     7081 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
     7141 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
     7201 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
     7261 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
     7321 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
     7381 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
     7441 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     7501 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     7561 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7621 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     7681 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     7741 tcagagagtg agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt
     7801 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7861 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7921 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     8101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8641 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8821 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8881 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8941 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9001 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9061 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9121 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9181 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9241 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9301 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9361 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9421 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9481 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9541 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9601 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9661 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9721 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9781 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9961 acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat
    10021 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    10081 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta
    10141 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt
    10201 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
    10261 gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac
    10321 tacaaactta gtagt
//
