LOCUS       V047662                 7714 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047662
VERSION     V047662
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"
     CDS             964..3204
                     /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             3211..3276
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3314..3538
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3584..4012
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4026..4355
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4422..5216
                     /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    5390..5511
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5560..5576)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5584..5600
                     /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(5608..5638)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5653..5674
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5962..6547)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6718..7578)
                     /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(7579..7683)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcgg acgaggcggc cctcgccctt cagcccggcg gctccccctc ggcggcgggg
     1021 gccgacaggg aggccgcgtc gtcccccgcc ggggagccgc tccgcaagag gccgcggaga
     1081 gatggtcccg gcctcgagcg gagcccgggc gagcccggtg gggcggcccc agagcgtgag
     1141 gtgccggcgg cggccagggg ctgcccgggt gcggcggcgg cggcgctgtg gcgggaggcg
     1201 gaggcagagg cggcggcggc aggcggggag caagaggccc aggcgactgc ggcggctggg
     1261 gaaggagaca atgggccggg cctgcagggc ccatctcggg agccaccgct ggccgacaac
     1321 ttgtacgacg aagacgacga cgacgagggc gaggaggagg aagaggcggc ggcggcggcg
     1381 attgggtacc gagataacct tctgttcggt gatgaaatta tcactaatgg ttttcattcc
     1441 tgtgaaagtg atgaggagga tagagcctca catgcaagct ctagtgactg gactccaagg
     1501 ccacggatag gtccatatac ttttgttcag caacatctta tgattggcac agatcctcga
     1561 acaattctta aagatttatt gccggaaaca atacctccac ctgagttgga tgatatgaca
     1621 ctgtggcaga ttgttattaa tatcctttca gaaccaccaa aaaggaaaaa aagaaaagat
     1681 attaatacaa ttgaagatgc tgtgaaatta ctgcaagagt gcaaaaaaat tatagttcta
     1741 actggagctg gggtgtctgt ttcatgtgga atacctgact tcaggtcaag ggatggtatt
     1801 tatgctcgcc ttgctgtaga cttcccagat cttccagatc ctcaagcgat gtttgatatt
     1861 gaatatttca gaaaagatcc aagaccattc ttcaagtttg caaaggaaat atatcctgga
     1921 caattccagc catctctctg tcacaaattc atagccttgt cagataagga aggaaaacta
     1981 cttcgcaact atacccagaa catagacacg ctggaacagg ttgcgggaat ccaaaggata
     2041 attcagtgtc atggttcctt tgcaacagca tcttgcctga tttgtaaata caaagttgac
     2101 tgtgaagctg tacgaggaga tatttttaat caggtagttc ctcgatgtcc taggtgccca
     2161 gctgatgaac cgcttgctat catgaaacca gagattgtgt tttttggtga aaatttacca
     2221 gaacagtttc atagagccat gaagtatgac aaagatgaag ttgacctcct cattgttatt
     2281 gggtcttccc tcaaagtaag accagtagca ctaattccaa gttccatacc ccatgaagtg
     2341 cctcagatat taattaatag agaacctttg cctcatctgc attttgatgt agagcttctt
     2401 ggagactgtg atgtcataat taatgaattg tgtcataggt taggtggtga atatgccaaa
     2461 ctttgctgta accctgtaaa gctttcagaa attactgaaa aacctccacg aacacaaaaa
     2521 gaattggctt atttgtcaga gttgccaccc acacctcttc atgtttcaga agactcaagt
     2581 tcaccagaaa gaacttcacc accagattct tcagtgattg tcacactttt agaccaagca
     2641 gctaagagta atgatgattt agatgtgtct gaatcaaaag gttgtatgga agaaaaacca
     2701 caggaagtac aaacttctag gaatgttgaa agtattgctg aacagatgga aaatccggat
     2761 ttgaagaatg ttggttctag tactggggag aaaaatgaaa gaacttcagt ggctggaaca
     2821 gtgagaaaat gctggcctaa tagagtggca aaggagcaga ttagtaggcg gcttgatggt
     2881 aatcagtatc tgtttttgcc accaaatcgt tacattttcc atggcgctga ggtatattca
     2941 gactctgaag atgacgtctt atcctctagt tcttgtggca gtaacagtga tagtgggaca
     3001 tgccagagtc caagtttaga agaacccatg gaggatgaaa gtgaaattga agaattctac
     3061 aatggcttag aagatgagcc tgatgttcca gagagagctg gaggagctgg atttgggact
     3121 gatggagatg atcaagaggc aattaatgaa gctatatctg tgaaacagga agtaacagac
     3181 atgaactatc catcaaacaa atcactcgag gactacaaag accatgacgg tgattataaa
     3241 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     3301 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3361 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3421 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3481 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3541 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3661 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3781 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3901 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3961 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4021 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4081 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4141 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4201 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4261 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4321 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4381 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4441 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4501 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4561 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4621 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4681 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4741 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4801 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4861 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4921 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4981 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5041 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5101 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5161 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5221 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5281 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5341 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5521 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5581 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5641 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5701 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5761 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6361 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6481 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6541 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6601 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6661 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6721 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6781 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6841 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6901 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6961 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7021 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7081 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7141 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7201 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7261 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7321 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7381 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7441 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7501 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7561 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7621 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7681 gcgcacattt ccccgaaaag tgccacctga cgtc
//
