LOCUS       V043147                 7407 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043147
VERSION     V043147
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2087
                     /label="Sirt1(NM_001372090)"
                     /note="Sirt1(NM_001372090)"
                     /gene="Sirt1"
     CDS             2094..2159
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2183..2771
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2837..2885
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3087..3515
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3529..3858
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3925..4719
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4895..5016
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5065..5081)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5089..5105
                     /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(5113..5143)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5158..5179
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5467..6055)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6226..7086)
                     /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(7087..7191)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtg cctgtgcagt ggaaggaaag
      781 caattttgga aatatatccc ggacagttcc agccatctct gtgtcacaaa ttcatagctt
      841 tgtcagataa ggaaggaaaa ctacttcgaa attatactca aaatatagat accttggagc
      901 aggttgcagg aatccaaagg atcattcagt gtcatggttc ctttgcaaca gcatcttgcc
      961 tgatttgtaa atacaaagtt gattgtgaag ctgttcgtgg agatattttt aatcaggtag
     1021 ttcctcggtg tcctaggtgc ccagctgatg agccacttgc catcatgaag ccagagattg
     1081 tcttctttgg tgaaaactta ccagaacagt ttcatagagc catgaagtat gacaaagatg
     1141 aagttgacct cctcattgtt attgggtctt ctctgaaagt aagaccagta gcactaattc
     1201 caagttctat accccatgaa gtgcctcaaa tattaataaa tagggaacct ctgcctcatc
     1261 tacattttga tgtagagctt cttggagact gcgatgtcat aattaatgag ttgtgtcata
     1321 ggttaggtgg cgagtatgcc aaactttgtt gtaaccctgt aaagctttca gaaattactg
     1381 aaaaacctcc acgaacacaa aaggaattgg ttcatttatc agagttgcca ccaacacctc
     1441 ttcatatttc agaagactca agttcacctg aaagaactgt accacaagac tcttctgtga
     1501 ttgctacact tgtagaccaa acaataaaga acaaagttga cgatttagaa gtatctgaac
     1561 caaaaagttg tgtggaagaa aaatcacagg aagtacagac ttataggaat gttgagagca
     1621 ttaatgtgga aaacccagat ttcaaggctg ttggttccag tactggagac aaaaatgaaa
     1681 gaacttctgt tgcagaaaca gtgagaaaat gctggcctaa tagacttgca aaggagcaga
     1741 ttagtaagcg tcttgacggt aatcaatact tgtttgtacc accaaatcgt tacatatttc
     1801 atggtgctga ggtatactca gactctgaag atgacgcctt atcctctagt tcctgtggca
     1861 gtaacagtga cagtggcaca tgccagagtc caagtttaga agaacccttg gaagatgaaa
     1921 gtgaaattga agagttctac aatggcttgg aagatgatgc tgacagaccg gagtgtgctg
     1981 gaggatctgg agctgacgga ggggatcaag aggcagttaa tgaagctata gccatgaaac
     2041 aggaattaac agatgtaaac tgtacaccag acaaatcaga gcactatgga tccgactaca
     2101 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2161 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2221 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2281 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2341 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2401 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2461 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2521 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2581 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2641 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2701 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2761 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     2821 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     2881 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     2941 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3001 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3061 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3121 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3181 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3241 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3301 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3361 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3421 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3481 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3541 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3601 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3661 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3781 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3841 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3901 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3961 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4021 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4081 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4141 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4201 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4261 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4321 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4381 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4441 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4501 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4561 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4621 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4681 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     4741 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4801 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4861 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4921 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4981 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5041 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5101 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5161 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5221 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5281 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5341 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5401 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5461 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5521 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5581 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5641 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5701 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5761 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5821 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5881 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5941 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6001 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6061 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6121 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6181 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6241 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6301 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6361 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6421 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6481 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6541 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6601 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6661 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6721 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6781 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6841 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6901 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6961 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7021 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7081 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7141 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7201 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7261 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7321 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7381 gacaattgca tgaagaatct gcttagg
//
