LOCUS       V024474                10021 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024474
VERSION     V024474
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..4947
                     /label="CDH1(NM_004360)"
                     /note="CDH1(NM_004360)"
                     /gene="CDH1"
     promoter        4991..5202
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5215..5483
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5510..6109
                     /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    6116..6704
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6778..7011
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7083..7204
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7223..7358
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7396..7412)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7420..7436
                     /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(7444..7474)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7489..7510
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7798..8386)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8557..9417)
                     /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(9418..9522)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9996..10012
                     /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 catgggccct tggagccgca gcctctcggc gctgctgctg
     2341 ctgctgcagg tctcctcttg gctctgccag gagccggagc cctgccaccc tggctttgac
     2401 gccgagagct acacgttcac ggtgccccgg cgccacctgg agagaggccg cgtcctgggc
     2461 agagtgaatt ttgaagattg caccggtcga caaaggacag cctatttttc cctcgacacc
     2521 cgattcaaag tgggcacaga tggtgtgatt acagtcaaaa ggcctctacg gtttcataac
     2581 ccacagatcc atttcttggt ctacgcctgg gactccacct acagaaagtt ttccaccaaa
     2641 gtcacgctga atacagtggg gcaccaccac cgccccccgc cccatcaggc ctccgtttct
     2701 ggaatccaag cagaattgct cacatttccc aactcctctc ctggcctcag aagacagaag
     2761 agagactggg ttattcctcc catcagctgc ccagaaaatg aaaaaggccc atttcctaaa
     2821 aacctggttc agatcaaatc caacaaagac aaagaaggca aggttttcta cagcatcact
     2881 ggccaaggag ctgacacacc ccctgttggt gtctttatta ttgaaagaga aacaggatgg
     2941 ctgaaggtga cagagcctct ggatagagaa cgcattgcca catacactct cttctctcac
     3001 gctgtgtcat ccaacgggaa tgcagttgag gatccaatgg agattttgat cacggtaacc
     3061 gatcagaatg acaacaagcc cgaattcacc caggaggtct ttaaggggtc tgtcatggaa
     3121 ggtgctcttc caggaacctc tgtgatggag gtcacagcca cagacgcgga cgatgatgtg
     3181 aacacctaca atgccgccat cgcttacacc atcctcagcc aagatcctga gctccctgac
     3241 aaaaatatgt tcaccattaa caggaacaca ggagtcatca gtgtggtcac cactgggctg
     3301 gaccgagaga gtttccctac gtataccctg gtggttcaag ctgctgacct tcaaggtgag
     3361 gggttaagca caacagcaac agctgtgatc acagtcactg acaccaacga taatcctccg
     3421 atcttcaatc ccaccacgta caagggtcag gtgcctgaga acgaggctaa cgtcgtaatc
     3481 accacactga aagtgactga tgctgatgcc cccaataccc cagcgtggga ggctgtatac
     3541 accatattga atgatgatgg tggacaattt gtcgtcacca caaatccagt gaacaacgat
     3601 ggcattttga aaacagcaaa gggcttggat tttgaggcca agcagcagta cattctacac
     3661 gtagcagtga cgaatgtggt accttttgag gtctctctca ccacctccac agccaccgtc
     3721 accgtggatg tgctggatgt gaatgaagcc cccatctttg tgcctcctga aaagagagtg
     3781 gaagtgtccg aggactttgg cgtgggccag gaaatcacat cctacactgc ccaggagcca
     3841 gacacattta tggaacagaa aataacatat cggatttgga gagacactgc caactggctg
     3901 gagattaatc cggacactgg tgccatttcc actcgggctg agctggacag ggaggatttt
     3961 gagcacgtga agaacagcac gtacacagcc ctaatcatag ctacagacaa tggttctcca
     4021 gttgctactg gaacagggac acttctgctg atcctgtctg atgtgaatga caacgccccc
     4081 ataccagaac ctcgaactat attcttctgt gagaggaatc caaagcctca ggtcataaac
     4141 atcattgatg cagaccttcc tcccaataca tctcccttca cagcagaact aacacacggg
     4201 gcgagtgcca actggaccat tcagtacaac gacccaaccc aagaatctat cattttgaag
     4261 ccaaagatgg ccttagaggt gggtgactac aaaatcaatc tcaagctcat ggataaccag
     4321 aataaagacc aagtgaccac cttagaggtc agcgtgtgtg actgtgaagg ggccgctggc
     4381 gtctgtagga aggcacagcc tgtcgaagca ggattgcaaa ttcctgccat tctggggatt
     4441 cttggaggaa ttcttgcttt gctaattctg attctgctgc tcttgctgtt tcttcggagg
     4501 agagcggtgg tcaaagagcc cttactgccc ccagaggatg acacccggga caacgtttat
     4561 tactatgatg aagaaggagg cggagaagag gaccaggact ttgacttgag ccagctgcac
     4621 aggggcctgg acgctcggcc tgaagtgact cgtaacgacg ttgcaccaac cctcatgagt
     4681 gtcccccggt atcttccccg ccctgccaat cccgatgaaa ttggaaattt tattgatgaa
     4741 aatctgaaag cggctgatac tgaccccaca gccccgcctt atgattctct gctcgtgttt
     4801 gactatgaag gaagcggttc cgaagctgct agtctgagct ccctgaactc ctcagagtca
     4861 gacaaagacc aggactatga ctacttgaac gaatggggca atcgcttcaa gaagctggct
     4921 gacatgtacg gaggcggcga ggacgactag cggccgcgaa ggatctgcga tcgctccggt
     4981 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     5041 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     5101 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     5161 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     5221 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     5281 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     5341 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     5401 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     5461 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta
     5521 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     5581 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     5641 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5701 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5761 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5821 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5881 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5941 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     6001 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     6061 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca
     6121 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6181 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6241 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6301 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6361 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6421 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6481 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6541 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6601 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6661 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6721 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6781 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6841 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6901 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6961 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7021 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7081 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7141 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7201 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     7261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     7321 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     7381 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7441 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7501 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8221 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8521 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8581 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8641 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8701 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8761 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8821 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8881 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8941 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9001 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9061 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9121 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9181 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9241 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9301 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9361 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9421 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9541 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9601 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9661 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9721 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9781 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9841 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9901 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9961 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
    10021 g
//
