LOCUS       V019390                 8207 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019390
VERSION     V019390
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        431..708
                     /label="chicken β-actin promoter"
     intron          709..1726
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     CDS             1823..2887
                     /label="SIRT6(NM_016539)"
                     /note="SIRT6(NM_016539)"
                     /gene="SIRT6"
     CDS             2894..2959
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2983..3571
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3637..3685
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3887..4315
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4329..4658
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4725..5519
                     /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    5695..5816
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5865..5881)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5889..5905
                     /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(5913..5943)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5958..5979
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6267..6855)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7026..7886)
                     /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(7887..7991)
                     /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 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      481 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      541 gggggggggg ggcccccccc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      601 cggaaaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      661 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgttgcg
      721 cgctgccttc cccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      781 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      841 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      901 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      961 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     1021 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1081 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1141 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1201 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1261 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1321 gccggggagg gctcggggga aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
     1381 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
     1441 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1501 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
     1561 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1621 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1681 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1741 gctggttatt gtgctgtctc atcattttgg caaagaattc tgcagctggc tagcgtttaa
     1801 acttaagctt ggtaccgcca ccatgtcggt gaattacgcg gcggggctgt cgccgtacgc
     1861 ggacaagggc aagtgcggcc tcccggagat cttcgacccc ccggaggagc tggagcggaa
     1921 ggtgtgggaa ctggcgaggc tggtctggca gtcttccagt gtggtgttcc acacgggtgc
     1981 cggcatcagc actgcctctg gcatccccga cttcaggggt ccccacggag tctggaccat
     2041 ggaggagcga ggtctggccc ccaagttcga caccaccttt gagagcgcgc ggcccacgca
     2101 gacccacatg gcgctggtgc agctggagcg cgtgggcctc ctccgcttcc tggtcagcca
     2161 gaacgtggac gggctccatg tgcgctcagg cttccccagg gacaaactgg cagagctcca
     2221 cgggaacatg tttgtggaag aatgtgccaa gtgtaagacg cagtacgtcc gagacacagt
     2281 cgtgggcacc atgggcctga aggccacggg ccggctctgc accgtggcta aggcaagggg
     2341 gctgcgagcc tgcaggggag agctgaggga caccatccta gactgggagg actccctgcc
     2401 cgaccgggac ctggcactcg ccgatgaggc cagcaggaac gccgacctgt ccatcacgct
     2461 gggtacatcg ctgcagatcc ggcccagcgg gaacctgccg ctggctacca agcgccgggg
     2521 aggccgcctg gtcatcgtca acctgcagcc caccaagcac gaccgccatg ctgacctccg
     2581 catccatggc tacgttgacg aggtcatgac ccggctcatg aagcacctgg ggctggagat
     2641 ccccgcctgg gacggccccc gtgtgctgga gagggcgctg ccacccctgc cccgcccgcc
     2701 cacccccaag ctggagccca aggaggaatc tcccacccgg atcaacggct ctatccccgc
     2761 cggccccaag caggagccct gcgcccagca caacggctca gagcccgcca gccccaaacg
     2821 ggagcggccc accagccctg ccccccacag accccccaaa agggtgaagg ccaaggcggt
     2881 ccccagcctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     2941 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     3001 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3061 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3121 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3181 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3241 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3301 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3361 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     3421 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3481 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3541 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     3601 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3661 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3841 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     3901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4021 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     4081 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4201 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     4261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4321 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4561 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4621 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4681 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4741 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4801 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4861 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4921 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4981 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5041 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5101 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5161 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5221 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5281 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5341 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5401 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5461 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5521 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5581 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5641 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5821 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5881 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5941 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6001 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7981 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8041 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8101 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8161 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
