LOCUS       V019919                 8243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019919
VERSION     V019919
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"
     CDS             683..2923
                     /codon_start=1
                     /gene=""
                     /note="homolog of budding yeast Sir2"
                     /product="human NAD+-dependent protein deacetylase
                     sirtuin-1"
                     /protein_id=""
                     /transl_table=1
                     /translation="MADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLE
                     RSPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNG
                     PGLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESD
                     EEDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQ
                     IVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYA
                     RLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLL
                     RNYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCP
                     ADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHE
                     VPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRT
                     QKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCME
                     EKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISR
                     RLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESE
                     IEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
                     /label="SIRT1"
     CDS             2930..2995
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3019..3607
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3673..3721
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3923..4351
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4365..4694
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4761..5555
                     /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    5731..5852
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5901..5917)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5925..5941
                     /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(5949..5979)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5994..6015
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6303..6891)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7062..7922)
                     /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(7923..8027)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcgga cgaggcggcc ctcgcccttc agcccggcgg
      721 ctccccctcg gcggcggggg ccgacaggga ggccgcgtcg tcccccgccg gggagccgct
      781 ccgcaagagg ccgcggagag atggtcccgg cctcgagcgg agcccgggcg agcccggtgg
      841 ggcggcccca gagcgtgagg tgccggcggc ggccaggggc tgcccgggtg cggcggcggc
      901 ggcgctgtgg cgggaggcgg aggcagaggc ggcggcggca ggcggggagc aagaggccca
      961 ggcgactgcg gcggctgggg aaggagacaa tgggccgggc ctgcagggcc catctcggga
     1021 gccaccgctg gccgacaact tgtacgacga agacgacgac gacgagggcg aggaggagga
     1081 agaggcggcg gcggcggcga ttgggtaccg agataacctt ctgttcggtg atgaaattat
     1141 cactaatggt tttcattcct gtgaaagtga tgaggaggat agagcctcac atgcaagctc
     1201 tagtgactgg actccaaggc cacggatagg tccatatact tttgttcagc aacatcttat
     1261 gattggcaca gatcctcgaa caattcttaa agatttattg ccggaaacaa tacctccacc
     1321 tgagttggat gatatgacac tgtggcagat tgttattaat atcctttcag aaccaccaaa
     1381 aaggaaaaaa agaaaagata ttaatacaat tgaagatgct gtgaaattac tgcaagagtg
     1441 caaaaaaatt atagttctaa ctggagctgg ggtgtctgtt tcatgtggaa tacctgactt
     1501 caggtcaagg gatggtattt atgctcgcct tgctgtagac ttcccagatc ttccagatcc
     1561 tcaagcgatg tttgatattg aatatttcag aaaagatcca agaccattct tcaagtttgc
     1621 aaaggaaata tatcctggac aattccagcc atctctctgt cacaaattca tagccttgtc
     1681 agataaggaa ggaaaactac ttcgcaacta tacccagaac atagacacgc tggaacaggt
     1741 tgcgggaatc caaaggataa ttcagtgtca tggttccttt gcaacagcat cttgcctgat
     1801 ttgtaaatac aaagtagatt gcgaggcagt ccgaggagat atttttaatc aggtagttcc
     1861 tcgatgtcct aggtgcccag ctgatgaacc gcttgctatc atgaaaccag agattgtgtt
     1921 ttttggtgaa aatttaccag aacagtttca tagagccatg aagtatgaca aagatgaagt
     1981 tgacctcctc attgttattg ggtcttccct caaagtaaga ccagtagcac taattccaag
     2041 ttccataccc catgaagtgc ctcagatatt aattaataga gaacctttgc ctcatctgca
     2101 ttttgatgta gagcttcttg gagactgtga tgtcataatt aatgaattgt gtcataggtt
     2161 aggtggtgaa tatgccaaac tttgctgtaa ccctgtaaag ctttcagaaa ttactgaaaa
     2221 acctccacga acacaaaaag aattggctta tttgtcagag ttgccaccca cacctcttca
     2281 tgtttcagaa gactcaagtt caccagaaag aacttcacca ccagattctt cagtgattgt
     2341 cacactttta gaccaagcag ctaagagtaa tgatgattta gatgtgtctg aatcaaaagg
     2401 ttgtatggaa gaaaaaccac aggaagtaca aacttctagg aatgttgaaa gtattgctga
     2461 acagatggaa aatccggatt tgaagaatgt tggttctagt actggggaga aaaatgaaag
     2521 aacttcagtg gctggaacag tgagaaaatg ctggcctaat agagtggcaa aggagcagat
     2581 tagtaggcgg cttgatggta atcagtatct gtttttgcca ccaaatcgtt acattttcca
     2641 tggcgctgag gtatattcag actctgaaga tgacgtctta tcctctagtt cttgtggcag
     2701 taacagtgat agtgggacat gccagagtcc aagtttagaa gaacccatgg aggatgaaag
     2761 tgaaattgaa gaattctaca atggcttaga agatgagcct gatgttccag agagagctgg
     2821 aggagctgga tttgggactg atggagatga tcaagaggca attaatgaag ctatatctgt
     2881 gaaacaggaa gtaacagaca tgaactatcc atcaaacaaa tcactcgagg actacaaaga
     2941 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     3001 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3061 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3121 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3181 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3241 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3301 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3361 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3421 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3481 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3541 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3601 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3661 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3721 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3781 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3841 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3901 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3961 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4021 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4081 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4141 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4201 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4261 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4321 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4381 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4441 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4501 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4561 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4621 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4681 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4741 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4801 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4861 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4921 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4981 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5041 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5101 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5161 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5221 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5281 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5341 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5401 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5461 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5521 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5581 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5641 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5701 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5881 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6481 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6721 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8041 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     8101 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8161 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     8221 attgcatgaa gaatctgctt agg
//
