LOCUS       V043443                 6466 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043443
VERSION     V043443
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             931..1902
                     /label="SIAH2(NM_005067)"
                     /note="SIAH2(NM_005067)"
                     /gene="SIAH2"
     CDS             1909..1998
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2033..2257
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2303..2731
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2745..3074
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3141..3935
                     /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    4109..4230
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4279..4295)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4303..4319
                     /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(4327..4357)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4372..4393
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4681..5266)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5437..6297)
                     /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(6298..6402)
                     /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 cactagtcca gtgtggtgga attcgccacc atgagccgcc cgtcctccac cggccccagc
      961 gctaataaac cctgcagcaa gcagccgccg ccgcagcccc agcacactcc gtccccggct
     1021 gcgcccccgg ccgccgccac catctcggct gcgggccccg gctcgtccgc ggtgcccgcc
     1081 gcggcggcgg tgatctcggg ccccggcggc ggcggcgggg ccggcccggt gtccccgcag
     1141 caccacgagc tgacctcgct cttcgagtgt ccggtctgct ttgactatgt cctgcctcct
     1201 attctgcagt gccaggccgg gcacctggtg tgtaaccaat gccgccagaa gttgagctgc
     1261 tgcccgacgt gcaggggcgc cctgacgccc agcatcagga acctggctat ggagaaggtg
     1321 gcctcggcag tcctgtttcc ctgtaagtat gccaccacgg gctgttccct gaccctgcac
     1381 catacggaga aaccagaaca tgaagacata tgtgaatacc gtccctactc ctgcccatgt
     1441 cctggtgctt cctgcaagtg gcaggggtcc ctggaagctg tgatgtccca tctcatgcac
     1501 gcccacaaga gcattaccac ccttcaggga gaagacatcg tctttctagc tacagacatt
     1561 aacttgccag gggctgtcga ctgggtgatg atgcagtcat gttttggcca tcacttcatg
     1621 ctggtgctgg agaaacaaga gaagtacgaa ggccaccagc agttttttgc catcgtcctg
     1681 ctcattggca cccgcaagca agccgagaac tttgcctaca gactggagtt gaatgggaac
     1741 cggcggagat tgacctggga ggccacgccc cgttcgattc atgacggtgt ggctgcggcc
     1801 atcatgaaca gcgactgcct tgttttcgac acagccatag cacatctttt tgcagataat
     1861 gggaaccttg gaatcaatgt tactatttct acatgttgtc cactcgagga acaaaaacta
     1921 atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga gcaaaagtta
     1981 atttccgagg aggacttatg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2041 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2101 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2161 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2221 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2281 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2341 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2401 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2461 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2521 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2581 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2641 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2701 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2821 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2881 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2941 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3001 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3061 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3121 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3181 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3241 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3301 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3361 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3421 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3481 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3541 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3601 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3661 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3721 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3781 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3841 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3901 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3961 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4021 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4081 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4141 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4201 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4261 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4321 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4381 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4441 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4501 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4561 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4621 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4681 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4741 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4801 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4861 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4921 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4981 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5041 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5101 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5161 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     5221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6421 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
