LOCUS       V019796                 8264 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019796
VERSION     V019796
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..2956
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2960..2986
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2990..3016
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3040..3628
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3694..3742
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3944..4372
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4386..4715
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4782..5576
                     /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    5752..5873
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5922..5938)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5946..5962
                     /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(5970..6000)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6015..6036
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6324..6912)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7083..7943)
                     /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(7944..8048)
                     /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 aaagttgact gtgaagctgt acgaggagat 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 tcactcgagt acccatacga
     2941 cgtacctgat tatgcaactt atccttatga tgtacctgat tatgctagct atccttatga
     3001 tgtcccagac tacgcatagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     3061 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3121 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3181 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3241 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3301 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3361 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3421 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3481 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3541 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3601 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     3661 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     3721 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3781 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3841 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3901 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3961 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4021 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4081 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     4141 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4201 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4261 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     4321 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4381 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4441 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4561 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4621 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4681 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4741 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4801 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4861 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4921 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4981 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5041 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5101 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5161 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5221 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5281 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5341 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5401 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5461 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5521 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5581 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5641 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5701 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5881 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8041 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     8101 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     8161 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     8221 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
