LOCUS       V034248                 6901 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034248
VERSION     V034248
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1557
                     /label="ICMT(NM_012405)"
                     /note="ICMT(NM_012405)"
                     /gene="ICMT"
     CDS             1564..1653
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    1677..2265
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2331..2379
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2581..3009
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3023..3352
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3419..4213
                     /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    4389..4510
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4559..4575)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4583..4599
                     /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(4607..4637)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4652..4673
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4961..5549)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5720..6580)
                     /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(6581..6685)
                     /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 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gggctgcgcg
      721 gcgcgggctc cgccgggctc tgaggcgcgt ctcagcctcg ccaccttcct gctgggcgcc
      781 tcggtgctcg cgctgccgct gctcacgcgc gccggcctgc agggccgcac cgggctggcg
      841 ctctacgtgg ccgggctcaa cgcgctgctg ctgctgctct atcggccgcc tcgctaccag
      901 atagccatcc gagcttgttt cctggggttt gtgttcggct gcggcacgct gctaagtttt
      961 agccagtctt cttggagtca ctttggctgg tacatgtgct ccctgtcatt gttccactat
     1021 tctgaatact tggtgacagc agtcaataat cccaaaagtc tgtccttgga ttcctttctc
     1081 ctgaatcaca gcctggagta tacagtagct gctctttctt cttggttaga gttcacactt
     1141 gaaaatatct tttggccaga actgaagcag attacctggc tcagtgtcac agggctgctg
     1201 atggtggtct tcggagaatg tctgaggaag gcggccatgt ttacagctgg ctccaatttc
     1261 aaccacgtgg tacagaatga aaaatcagat acacatactc tggtgaccag tggagtgtac
     1321 gcttggtttc ggcatccttc ttacgtcggg tggttttact ggagtattgg aactcaggtg
     1381 atgctgtgta accccatctg cggcgtcagc tatgccctga cagtgtggcg attcttccgc
     1441 gatcgaacag aagaagaaga aatctcacta attcactttt ttggagagga gtacctggag
     1501 tataagaaga gggtgcccac gggcctgcct ttcataaagg gggtcaaggt ggacctgctc
     1561 gaggaacaaa aactaatatc agaggaagat ttggaacaga aactaatcag tgaagaagat
     1621 ttagagcaaa agttaatttc cgaggaggac ttatagtgag ggcccgatat ctcgacaatc
     1681 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1741 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1801 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     1861 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     1921 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     1981 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2041 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     2101 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2161 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2221 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     2281 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     2341 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     2401 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     2461 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     2521 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     2581 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2641 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2701 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     2761 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2821 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2881 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2941 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3001 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3181 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3241 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3301 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3361 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3421 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3481 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3541 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3601 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3661 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3721 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3781 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3841 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3901 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3961 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4021 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4081 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4141 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4201 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     4261 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4321 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4381 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4441 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4501 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4561 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4621 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4681 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4741 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     5161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5401 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6661 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6721 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     6781 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     6841 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     6901 g
//
