LOCUS       V045265                 9311 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045265
VERSION     V045265
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"
     regulatory      2296..2305
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2305..2346
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             2353..4230
                     /label="CANX(NM_001363995)"
                     /note="CANX(NM_001363995)"
                     /gene="CANX"
     promoter        4281..4492
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4505..4773
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4800..5399
                     /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    5406..5994
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6068..6301
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6373..6494
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6513..6648
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6686..6702)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6710..6726
                     /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(6734..6764)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6779..6800
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7088..7676)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7847..8707)
                     /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(8708..8812)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9286..9302
                     /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 catgggtaag cctatcccta accctctcct cggtctcgat
     2341 tctacactcg agatggaagg gaagtggttg ctgtgtatgt tactggtgct tggaactgct
     2401 attgttgagg ctcatgatgg acatgatgat gatgtgattg atattgagga tgaccttgac
     2461 gatgtcattg aagaggtaga agactcaaaa ccagatacca ctgctcctcc ttcatctccc
     2521 aaggttactt acaaagctcc agttccaaca ggggaagtat attttgctga ttcttttgac
     2581 agaggaactc tgtcagggtg gattttatcc aaagccaaga aagacgatac cgatgatgaa
     2641 attgccaaat atgatggaaa gtgggaggta gaggaaatga aggagtcaaa gcttccaggt
     2701 gataaaggac ttgtgttgat gtctcgggcc aagcatcatg ccatctctgc taaactgaac
     2761 aagcccttcc tgtttgacac caagcctctc attgttcagt atgaggttaa tttccaaaat
     2821 ggaatagaat gtggtggtgc ctatgtgaaa ctgctttcta aaacaccaga actcaacctg
     2881 gatcagttcc atgacaagac cccttatacg attatgtttg gtccagataa atgtggagag
     2941 gactataaac tgcacttcat cttccgacac aaaaacccca aaacgggtat ctatgaagaa
     3001 aaacatgcta agaggccaga tgcagatctg aagacctatt ttactgataa gaaaacacat
     3061 ctttacacac taaaaagccg agggactaat catttatgtt tcgtgcataa ggaaatgggc
     3121 gttagcttcc ccaagacccc aggagccagg cattgttctt accccaaagc ctcagtcttg
     3181 aatccagata atagttttga aatactggtt gaccaatctg tggtgaatag tggaaatctg
     3241 ctcaatgaca tgactcctcc tgtaaatcct tcacgtgaaa ttgaggaccc agaagaccgg
     3301 aagcccgagg attgggatga aagaccaaaa atcccagatc cagaagctgt caagccagat
     3361 gactgggatg aagatgcccc tgctaagatt ccagatgaag aggccacaaa acccgaaggc
     3421 tggttagatg atgagcctga gtacgtacct gatccagacg cagagaaacc tgaggattgg
     3481 gatgaagaca tggatggaga atgggaggct cctcagattg ccaaccctag atgtgagtca
     3541 gctcctggat gtggtgtctg gcagcgacct gtgattgaca accccaatta taaaggcaaa
     3601 tggaagcctc ctatgattga caatcccagt taccagggaa tctggaaacc caggaaaata
     3661 ccaaatccag atttctttga agatctggaa cctttcagaa tgactccttt tagtgctatt
     3721 ggtttggagc tgtggtccat gacctctgac attttttttg acaactttat catttgtgct
     3781 gatcgaagaa tagttgatga ttgggccaat gatggatggg gcctgaagaa agctgctgat
     3841 ggggctgctg agccaggcgt tgtggggcag atgatcgagg cagctgaaga gcgcccgtgg
     3901 ctgtgggtag tctatattct aactgtagcc cttcctgtgt tcctggttat cctcttctgc
     3961 tgttctggaa agaaacagac cagtggtatg gagtataaga aaactgatgc acctcaaccg
     4021 gatgtgaagg aagaggaaga agagaaggaa gaggaaaagg acaagggaga tgaggaggag
     4081 gaaggagaag agaaacttga agagaaacag aaaagtgatg ctgaagaaga tggtggcact
     4141 gtcagtcaag aggaggaaga cagaaaacct aaagcagagg aggatgaaat tttgaacaga
     4201 tcaccaagaa acagaaagcc acgaagagag tgaggatccg cggccgcgaa ggatctgcga
     4261 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     4321 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     4381 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     4441 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     4501 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     4561 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     4621 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     4681 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     4741 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga
     4801 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     4861 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     4921 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4981 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5041 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5101 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5161 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5221 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5281 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5341 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     5401 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5461 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5521 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5581 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5641 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5701 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5761 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5821 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5881 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5941 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta
     6001 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6061 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     6121 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6181 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6241 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6301 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6361 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6421 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6481 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6541 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6601 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6661 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6721 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6781 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6841 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6901 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6961 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7021 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7081 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7141 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7201 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7261 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7321 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7381 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7441 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7501 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7561 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7621 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7681 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7741 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7801 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7861 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7921 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7981 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8041 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8101 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8161 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8221 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8281 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8341 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8401 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8461 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8521 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8581 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8641 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8701 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8761 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8821 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8881 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8941 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     9001 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     9061 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     9121 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     9181 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     9241 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     9301 gtgccaagct g
//
