LOCUS       V021173                 7753 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021173
VERSION     V021173
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1033..3276
                     /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
                     RNYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYRVDCEAVRGDIFNQVVPRCPRCP
                     ADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHE
                     VPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRT
                     QKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCME
                     EKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISR
                     RLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESE
                     IEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS*"
                     /label="SIRT1"
     polyA_signal    3353..3577
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3623..4051
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4065..4394
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4461..5255
                     /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    5429..5550
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5599..5615)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5623..5639
                     /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(5647..5677)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5692..5713
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6001..6586)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6757..7617)
                     /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(7618..7722)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttatggcgga cgaggcggcc ctcgcccttc agcccggcgg ctccccctcg
     1081 gcggcggggg ccgacaggga ggccgcgtcg tcccccgccg gggagccgct ccgcaagagg
     1141 ccgcggagag atggtcccgg cctcgagcgg agcccgggcg agcccggtgg ggcggcccca
     1201 gagcgtgagg tgccggcggc ggccaggggc tgcccgggtg cggcggcggc ggcgctgtgg
     1261 cgggaggcgg aggcagaggc ggcggcggca ggcggggagc aagaggccca ggcgactgcg
     1321 gcggctgggg aaggagacaa tgggccgggc ctgcagggcc catctcggga gccaccgctg
     1381 gccgacaact tgtacgacga agacgacgac gacgagggcg aggaggagga agaggcggcg
     1441 gcggcggcga ttgggtaccg agataacctt ctgttcggtg atgaaattat cactaatggt
     1501 tttcattcct gtgaaagtga tgaggaggat agagcctcac atgcaagctc tagtgactgg
     1561 actccaaggc cacggatagg tccatatact tttgttcagc aacatcttat gattggcaca
     1621 gatcctcgaa caattcttaa agatttattg ccggaaacaa tacctccacc tgagttggat
     1681 gatatgacac tgtggcagat tgttattaat atcctttcag aaccaccaaa aaggaaaaaa
     1741 agaaaagata ttaatacaat tgaagatgct gtgaaattac tgcaagagtg caaaaaaatt
     1801 atagttctaa ctggagctgg ggtgtctgtt tcatgtggaa tacctgactt caggtcaagg
     1861 gatggtattt atgctcgcct tgctgtagac ttcccagatc ttccagatcc tcaagcgatg
     1921 tttgatattg aatatttcag aaaagatcca agaccattct tcaagtttgc aaaggaaata
     1981 tatcctggac aattccagcc atctctctgt cacaaattca tagccttgtc agataaggaa
     2041 ggaaaactac ttcgcaacta tacccagaac atagacacgc tggaacaggt tgcgggaatc
     2101 caaaggataa ttcagtgtca tggttccttt gcaacagcat cttgcctgat ttgtaaatac
     2161 agagttgact gtgaagctgt acgaggagat atttttaatc aggtagttcc tcgatgtcct
     2221 aggtgcccag ctgatgaacc gcttgctatc atgaaaccag agattgtgtt ttttggtgaa
     2281 aatttaccag aacagtttca tagagccatg aagtatgaca aagatgaagt tgacctcctc
     2341 attgttattg ggtcttccct caaagtaaga ccagtagcac taattccaag ttccataccc
     2401 catgaagtgc ctcagatatt aattaataga gaacctttgc ctcatctgca ttttgatgta
     2461 gagcttcttg gagactgtga tgtcataatt aatgaattgt gtcataggtt aggtggtgaa
     2521 tatgccaaac tttgctgtaa ccctgtaaag ctttcagaaa ttactgaaaa acctccacga
     2581 acacaaaaag aattggctta tttgtcagag ttgccaccca cacctcttca tgtttcagaa
     2641 gactcaagtt caccagaaag aacttcacca ccagattctt cagtgattgt cacactttta
     2701 gaccaagcag ctaagagtaa tgatgattta gatgtgtctg aatcaaaagg ttgtatggaa
     2761 gaaaaaccac aggaagtaca aacttctagg aatgttgaaa gtattgctga acagatggaa
     2821 aatccggatt tgaagaatgt tggttctagt actggggaga aaaatgaaag aacttcagtg
     2881 gctggaacag tgagaaaatg ctggcctaat agagtggcaa aggagcagat tagtaggcgg
     2941 cttgatggta atcagtatct gtttttgcca ccaaatcgtt acattttcca tggcgctgag
     3001 gtatattcag actctgaaga tgacgtctta tcctctagtt cttgtggcag taacagtgat
     3061 agtgggacat gccagagtcc aagtttagaa gaacccatgg aggatgaaag tgaaattgaa
     3121 gaattctaca atggcttaga agatgagcct gatgttccag agagagctgg aggagctgga
     3181 tttgggactg atggagatga tcaagaggca attaatgaag ctatatctgt gaaacaggaa
     3241 gtaacagaca tgaactatcc atcaaacaaa tcataggaat tctgcagata tccagcacag
     3301 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3361 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3421 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3481 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3541 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3601 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3661 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3841 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3901 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3961 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4021 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4141 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4201 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4261 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4321 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4381 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4441 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4501 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4561 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4621 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4681 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4741 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4801 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4861 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4921 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4981 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5041 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5101 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5161 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5221 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5281 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5341 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5401 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5461 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5521 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5581 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7621 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7681 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7741 gccacctgac gtc
//
