LOCUS       V044916                 8325 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044916
VERSION     V044916
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3148
                     /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             3185..3841
                     /codon_start=1
                     /gene=""
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /transl_table=1
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK*"
                     /label="GST"
     polyA_signal    3892..4116
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4162..4590
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4604..4933
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5000..5794
                     /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    5968..6089
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6138..6154)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6162..6178
                     /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(6186..6216)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6231..6252
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6540..7125)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7296..8156)
                     /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(8157..8261)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gcggacgagg cggccctcgc ccttcagccc ggcggctccc cctcggcggc
      961 gggggccgac agggaggccg cgtcgtcccc cgccggggag ccgctccgca agaggccgcg
     1021 gagagatggt cccggcctcg agcggagccc gggcgagccc ggtggggcgg ccccagagcg
     1081 tgaggtgccg gcggcggcca ggggctgccc gggtgcggcg gcggcggcgc tgtggcggga
     1141 ggcggaggca gaggcggcgg cggcaggcgg ggagcaagag gcccaggcga ctgcggcggc
     1201 tggggaagga gacaatgggc cgggcctgca gggcccatct cgggagccac cgctggccga
     1261 caacttgtac gacgaagacg acgacgacga gggcgaggag gaggaagagg cggcggcggc
     1321 ggcgattggg taccgagata accttctgtt cggtgatgaa attatcacta atggttttca
     1381 ttcctgtgaa agtgatgagg aggatagagc ctcacatgca agctctagtg actggactcc
     1441 aaggccacgg ataggtccat atacttttgt tcagcaacat cttatgattg gcacagatcc
     1501 tcgaacaatt cttaaagatt tattgccgga aacaatacct ccacctgagt tggatgatat
     1561 gacactgtgg cagattgtta ttaatatcct ttcagaacca ccaaaaagga aaaaaagaaa
     1621 agatattaat acaattgaag atgctgtgaa attactgcaa gagtgcaaaa aaattatagt
     1681 tctaactgga gctggggtgt ctgtttcatg tggaatacct gacttcaggt caagggatgg
     1741 tatttatgct cgccttgctg tagacttccc agatcttcca gatcctcaag cgatgtttga
     1801 tattgaatat ttcagaaaag atccaagacc attcttcaag tttgcaaagg aaatatatcc
     1861 tggacaattc cagccatctc tctgtcacaa attcatagcc ttgtcagata aggaaggaaa
     1921 actacttcgc aactataccc agaacataga cacgctggaa caggttgcgg gaatccaaag
     1981 gataattcag tgtcatggtt cctttgcaac agcatcttgc ctgatttgta aatacaaagt
     2041 tgactgtgaa gctgtacgag gagatatttt taatcaggta gttcctcgat gtcctaggtg
     2101 cccagctgat gaaccgcttg ctatcatgaa accagagatt gtgttttttg gtgaaaattt
     2161 accagaacag tttcatagag ccatgaagta tgacaaagat gaagttgacc tcctcattgt
     2221 tattgggtct tccctcaaag taagaccagt agcactaatt ccaagttcca taccccatga
     2281 agtgcctcag atattaatta atagagaacc tttgcctcat ctgcattttg atgtagagct
     2341 tcttggagac tgtgatgtca taattaatga attgtgtcat aggttaggtg gtgaatatgc
     2401 caaactttgc tgtaaccctg taaagctttc agaaattact gaaaaacctc cacgaacaca
     2461 aaaagaattg gcttatttgt cagagttgcc acccacacct cttcatgttt cagaagactc
     2521 aagttcacca gaaagaactt caccaccaga ttcttcagtg attgtcacac ttttagacca
     2581 agcagctaag agtaatgatg atttagatgt gtctgaatca aaaggttgta tggaagaaaa
     2641 accacaggaa gtacaaactt ctaggaatgt tgaaagtatt gctgaacaga tggaaaatcc
     2701 ggatttgaag aatgttggtt ctagtactgg ggagaaaaat gaaagaactt cagtggctgg
     2761 aacagtgaga aaatgctggc ctaatagagt ggcaaaggag cagattagta ggcggcttga
     2821 tggtaatcag tatctgtttt tgccaccaaa tcgttacatt ttccatggcg ctgaggtata
     2881 ttcagactct gaagatgacg tcttatcctc tagttcttgt ggcagtaaca gtgatagtgg
     2941 gacatgccag agtccaagtt tagaagaacc catggaggat gaaagtgaaa ttgaagaatt
     3001 ctacaatggc ttagaagatg agcctgatgt tccagagaga gctggaggag ctggatttgg
     3061 gactgatgga gatgatcaag aggcaattaa tgaagctata tctgtgaaac aggaagtaac
     3121 agacatgaac tatccatcaa acaaatcaga attcggtgga ggcggatcag gaggcggtgg
     3181 atctatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact
     3241 tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga
     3301 taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat
     3361 tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca
     3421 caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg aaggagcggt
     3481 tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg aaactctcaa
     3541 agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca
     3601 taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct
     3661 tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt
     3721 taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat
     3781 agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatg
     3841 agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     3901 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3961 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4021 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4081 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4141 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4201 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4261 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4321 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4381 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4441 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4501 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4561 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4621 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4681 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4741 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4801 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4861 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4921 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4981 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5041 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5101 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5161 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5221 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5281 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5341 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5401 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5461 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5521 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5581 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5641 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5701 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5761 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5821 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5881 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5941 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6001 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6061 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6121 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6181 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6241 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6301 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6361 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6421 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6481 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6541 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6601 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6661 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6721 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6781 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6841 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6901 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6961 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7021 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     7081 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7141 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7201 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7261 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7321 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7381 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7441 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7501 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7561 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7621 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7681 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7741 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7801 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7861 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7921 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7981 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8041 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8101 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8161 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8221 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8281 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
