LOCUS       V019181                 6172 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019181
VERSION     V019181
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             914..1660
                     /label="ING4(NM_001127582)"
                     /note="ING4(NM_001127582)"
                     /gene="ING4"
     CDS             1661..1684
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    1739..1963
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2009..2437
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2451..2780
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2847..3641
                     /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    3815..3936
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3985..4001)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4009..4025
                     /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(4033..4063)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4078..4099
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4387..4972)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5143..6003)
                     /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(6004..6108)
                     /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 cgctagcgcc accatggctg cggggatgta tttggaacat tatctggaca gtattgaaaa
      961 ccttcccttt gaattacaga gaaactttca gctcatgagg gacctagacc aaagaacaga
     1021 ggacctgaag gctgaaattg acaagttggc cactgagtat atgagtagtg cccgcagcct
     1081 gagctccgag gaaaaattgg cccttctcaa acagatccag gaagcctatg gcaagtgcaa
     1141 ggaatttggt gacgacaagg tgcagcttgc catgcagacc tatgagatgg tggacaaaca
     1201 cattcggcgg ctggacacag acctggcccg ttttgaggct gatctcaagg agaaacagat
     1261 tgagtcaagt gactatgaca gctcttccag caaaggcaaa aagaaaggcc ggactcaaaa
     1321 ggagaagaaa gctgctcgtg ctcgttccaa agggaaaaac tcggatgaag aagcccccaa
     1381 gactgcccag aagaagttaa agctcgtgcg cacaagtcct gagtatggga tgccctcagt
     1441 gacctttggc agtgtccacc cctctgatgt gttggatatg cctgtggatc ccaacgaacc
     1501 cacctattgc ctttgtcacc aggtctccta tggagagatg attggctgtg acaaccctga
     1561 ttgttccatt gagtggttcc attttgcctg tgtggggctg acaaccaagc ctcgggggaa
     1621 atggttttgc ccacgctgct cccaagaacg gaagaagaaa gattacaagg atgacgacga
     1681 taagtagggc gcgccctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     1741 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1801 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1861 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1921 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     1981 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2041 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2101 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2161 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2221 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2281 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2341 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2401 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2461 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2521 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2581 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2641 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2701 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2761 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2821 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2881 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2941 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3001 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3061 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3121 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3181 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3241 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3301 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3361 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3421 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3481 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3541 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3601 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3661 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3721 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3781 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3841 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3901 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3961 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4021 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4081 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4141 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4921 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6121 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt
//
