LOCUS       V018615                 6397 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018615
VERSION     V018615
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             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..1950
                     /label="SIRT4(NM_012240)"
                     /note="SIRT4(NM_012240)"
                     /gene="SIRT4"
     polyA_signal    1997..2221
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2267..2695
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2709..3038
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3105..3899
                     /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    4073..4194
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4243..4259)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4267..4283
                     /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(4291..4321)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4336..4357
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4645..5230)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5401..6261)
                     /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(6262..6366)
                     /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 ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat gaagatgagc
     1021 tttgcgttga ctttcaggtc agcaaaaggc cgttggatcg caaaccccag ccagccgtgc
     1081 tcgaaagcct ccattgggtt atttgtgcca gcaagtcctc ctctggaccc tgagaaggtc
     1141 aaagagttac agcgcttcat caccctttcc aagagactcc ttgtgatgac tggggcagga
     1201 atctccaccg aatcggggat accagactac aggtcagaaa aagtggggct ttatgcccgc
     1261 actgaccgca ggcccatcca gcatggtgat tttgtccgga gtgccccaat ccgccagcgg
     1321 tactgggcga gaaacttcgt aggctggcct caattctcct cccaccagcc taaccctgca
     1381 cactgggctt tgagcacctg ggagaaactc ggaaagctgt actggttggt gacccaaaat
     1441 gtggatgctt tgcacaccaa ggcggggagt cggcgcctga cagagctcca cggatgcatg
     1501 gacagggtcc tgtgcttgga ttgtggggaa cagactcccc ggggggtgct gcaagagcgt
     1561 ttccaagtcc tgaaccccac ctggagtgct gaggcccatg gcctggctcc tgatggtgac
     1621 gtctttctct cagaggagca agtccggagc tttcaggtcc caacctgcgt tcaatgtgga
     1681 ggccatctga aaccagatgt cgttttcttc ggggacacag tgaaccctga caaggttgat
     1741 tttgtgcaca agcgtgtaaa agaagccgac tccctcttgg tggtgggatc atccttgcag
     1801 gtatactctg gttacaggtt tatcctcact gcctgggaga agaagctccc gattgcaata
     1861 ctgaacattg ggcccacacg gtcggatgac ttggcgtgtc tgaaactgaa ttctcgttgt
     1921 ggagagttgc tgcctttgat agacccatgc tgactcgagt ctagagggcc cgtttaaacc
     1981 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2041 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2101 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2161 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2221 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2281 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2341 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2401 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2461 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2521 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2581 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2641 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2701 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2761 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2881 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2941 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3001 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3061 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3121 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3181 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3241 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3301 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3361 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3421 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3481 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3541 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3601 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3661 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3721 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3781 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3841 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3901 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3961 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4021 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4201 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5161 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6361 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
