LOCUS       V030686                10199 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030686
VERSION     V030686
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..5031
                     /label="ANO6(NM_001025356)"
                     /note="ANO6(NM_001025356)"
                     /gene="ANO6"
     CDS             5038..5064
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5068..5094
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5098..5124
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        5169..5380
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5393..5661
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5688..6287
                     /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    6294..6882
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6956..7189
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7261..7382
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7401..7536
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7574..7590)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7598..7614
                     /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(7622..7652)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7667..7688
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7976..8564)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8735..9595)
                     /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(9596..9700)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10174..10190
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgaaaaag atgagcagga atgttttgct acaaatggag
     2341 gaggaggagg acgacgacga tggggatatc gtgttggaaa accttggaca gacaattgtc
     2401 cccgatttgg gatcactgga aagtcagcat gattttcgaa ccccggagtt tgaagaattt
     2461 aatggaaaac ctgactccct cttttttaat gatggccagc gaagaattga ctttgttcta
     2521 gtatatgagg atgaaagcag aaaagagacc aataaaaagg gtacaaatga aaaacaaagg
     2581 aggaaaagac aagcatacga atctaacctt atctgtcatg gcctgcagtt agaagcaaca
     2641 agatcagtat tggatgacaa gcttgtattt gtaaaagtac acgcaccatg ggaggtgtta
     2701 tgtacgtatg ctgagataat gcacatcaaa ttgcctctga aacccaatga tctgaaaaac
     2761 cggtcctcag cctttggtac actcaactgg tttaccaaag tcctcagtgt agacgaaagc
     2821 atcatcaagc cagagcaaga gtttttcact gccccatttg agaagaaccg gatgaatgat
     2881 ttttacatag ttgatagaga tgctttcttc aatccagcca ccagaagccg cattgtttac
     2941 ttcatcctct ctcgggtcaa gtatcaagtg ataaacaatg ttagcaagtt tgggatcaac
     3001 agacttgtaa actctgggat ctacaaggca gctttcccac tccatgattg caaattccgc
     3061 cgtcagtcag aggatcccag ctgccctaat gaacggtacc ttctgtacag agaatgggct
     3121 catcctcgaa gcatatacaa aaagcagccc ttggatctta tcaggaaata ctatggagag
     3181 aagattggaa tctactttgc ttggctgggc tattacactc agatgcttct cctggccgca
     3241 gttgtaggag tggcttgctt tctctatgga tatcttaatc aagataactg tacatggagc
     3301 aaagaagttt gtcatcctga tattggtggc aagatcataa tgtgtcctca gtgtgatagg
     3361 ctttgtccat tctggaaact caatattact tgcgagtcct caaagaaatt gtgcatcttc
     3421 gacagttttg gaaccctggt ctttgcagta tttatgggag tatgggttac cttgtttttg
     3481 gagttttgga agcgacgcca ggcagaactt gagtatgaat gggatactgt tgagttacag
     3541 caggaagaac aagcccgacc agaatacgaa gcacgatgta ctcacgtagt gataaatgag
     3601 attactcagg aagaagaacg cattcccttt actgcctggg gaaaatgtat acggataacc
     3661 ctctgtgcca gtgctgtctt tttctggatc ctattgatca tcgcttcagt tattgggatc
     3721 attgtctata ggctctcggt gttcattgta ttttctgcaa aacttcccaa gaacattaat
     3781 ggaacagacc caatccagaa atacctgact ccacagacag ccacgtccat cacggcctcc
     3841 atcatcagct ttataattat catgattctg aacaccatat atgaaaaagt ggcaattatg
     3901 attactaact tcgaactccc aaggacccag actgattatg agaacagcct caccatgaag
     3961 atgttcttat tccagtttgt caactactac tcttcatgct tctacatagc attctttaag
     4021 ggcaaatttg taggctatcc aggagaccca gtttattggt tgggaaaata cagaaatgaa
     4081 gagtgtgacc caggtggctg tcttcttgaa ctgacaactc agctgacaat aatcatggga
     4141 ggaaaagcaa tctggaataa catacaagaa gtattattgc cctggatcat gaatctaatt
     4201 gggcgatttc acagagtttc tggatcagaa aagataaccc cacgatggga acaggactac
     4261 catctgcagc ctatgggcaa actgggatta ttttatgaat atcttgaaat gattattcag
     4321 tttgggttcg tcaccttatt tgtggcctct tttccactgg cccctctgtt ggctctcgtg
     4381 aacaatatat tggaaataag agtggacgca tggaaactga ccacccagtt tagacgcctg
     4441 gtaccagaga aagcccaaga cattggagca tggcagccca tcatgcaagg aatagcaatt
     4501 ctggctgtgg tgaccaatgc catgatcata gctttcacgt cggacatgat cccccgccta
     4561 gtgtactact ggtccttctc cgtccctccc tacggggacc acacttccta caccatggaa
     4621 gggtacatca acaacactct ctccatcttc aaagtcgcag acttcaaaaa caaaagcaag
     4681 ggaaacccgt actctgacct gggtaaccat accacatgca ggtatcgtga tttccgatac
     4741 ccacctggac acccccagga gtataaacac aacatctact attggcatgt gattgcagcc
     4801 aagctggctt ttatcattgt catggagcac gtcatctact ctgtgaaatt tttcatttca
     4861 tatgcaattc ccgatgtatc aaaacgcaca aagagcaaga tccagagaga aaaataccta
     4921 acccaaaagc ttcttcatga gaatcacctc aaagatatga cgaaaaatat gggggtgata
     4981 gctgagcgga tgatagaagc agtagataac aatttacggc caaaatcaga aggatcctac
     5041 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat
     5101 ccttatgatg tcccagacta cgcataggcg gccgcgaagg atctgcgatc gctccggtgc
     5161 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     5221 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     5281 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     5341 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     5401 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     5461 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     5521 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     5581 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     5641 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagatg accgagtaca
     5701 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
     5761 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
     5821 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5881 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     5941 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     6001 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     6061 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     6121 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     6181 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     6241 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc gacaatcaac
     6301 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6361 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6421 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6481 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6541 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6601 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6661 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     6721 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6781 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6841 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca
     6901 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     6961 gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg
     7021 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     7081 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     7141 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     7201 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     7261 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     7321 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     7381 tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc
     7441 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     7501 tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa
     7561 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7621 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7681 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7741 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7801 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7861 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7921 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7981 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8041 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8101 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8161 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8221 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8281 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8341 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8401 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8461 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     9001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     9061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     9121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9721 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     9781 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     9841 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     9901 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     9961 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
    10021 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
    10081 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
    10141 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
