LOCUS       V019119                 7231 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019119
VERSION     V019119
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             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1033..2784
                     /label="KRT10(NM_000421)"
                     /note="KRT10(NM_000421)"
                     /gene="KRT10"
     polyA_signal    2831..3055
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3101..3529
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3543..3872
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3939..4733
                     /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    4907..5028
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5077..5093)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5101..5117
                     /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(5125..5155)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5170..5191
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5479..6064)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6235..7095)
                     /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(7096..7200)
                     /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 ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttatgtctgt tcgatacagc tcaagcaagc actactcttc ctcccgcagt
     1081 ggaggaggag gaggaggagg aggatgtgga ggaggaggag gagtgtcatc cctaagaatt
     1141 tctagcagca aaggctccct tggtggagga tttagctcag gggggttcag tggtggctct
     1201 tttagccgtg ggagctctgg tgggggctgc tttgggggct catcaggtgg ctatggagga
     1261 ttaggaggtt ttggtggagg tagctttcgt ggaagctatg gaagtagcag ctttggtggg
     1321 agttatggag gcatctttgg agggggcagt ttcggaggtg gcagctttgg tgggggcagc
     1381 tttggtggag gcggctttgg tggaggcggc tttggaggag gctttggtgg tggatttgga
     1441 ggagatggtg gccttctctc tggaaatgaa aaagtaacca tgcagaatct gaatgaccgc
     1501 ctggcttcct acttggacaa agttcgggct ctggaagaat caaactatga gctggaaggc
     1561 aaaatcaagg agtggtatga aaagcatggc aactcacatc agggggagcc tcgtgactac
     1621 agcaaatact acaaaaccat cgatgacctt aaaaatcaga ttctcaacct aacaactgat
     1681 aatgccaaca tcctgcttca gatcgacaat gccaggctgg cagctgatga cttcaggctg
     1741 aagtatgaga atgaggtagc tctgcgccag agcgtggagg ctgacatcaa cggcctgcgt
     1801 agggtgctgg atgagctgac cctgaccaag gctgacctgg agatgcaaat tgagagcctg
     1861 actgaagagc tggcctatct gaagaagaac cacgaggagg aaatgaaaga ccttcgaaat
     1921 gtgtccactg gtgatgtgaa tgtggaaatg aatgctgccc cgggtgttga tctgactcaa
     1981 cttctgaata acatgagaag ccaatatgaa caacttgctg aacaaaaccg caaagatgct
     2041 gaagcctggt tcaatgaaaa gagcaaggaa ctgactacag aaattgataa taacattgaa
     2101 cagatatcca gctataaatc tgagattact gaattgagac gtaatgtaca agctctggag
     2161 atagaactac agtcccaact ggccttgaaa caatccctgg aagcctcctt ggcagaaaca
     2221 gaaggtcgct actgtgtgca gctctcacag attcaggccc agatatccgc tctggaagaa
     2281 cagttgcaac agattcgagc tgaaaccgag tgccagaata ctgaatacca acaactcctg
     2341 gatattaaga tccgactgga gaatgaaatt caaacctacc gcagcctgct agaaggagag
     2401 ggaagttccg gaggcggcgg acgcggcggc ggaagtttcg gcggcggcta cggcggcgga
     2461 agctccggcg gcggaagctc cggcggcggc cacggcggcg gccacggcgg cagttccggc
     2521 ggcggctacg gaggcggaag ctccggcggc ggaagctccg gcggcggcta cgggggcgga
     2581 agctccagcg gcggccacgg cggcagttcc agcggcggct acggtggtgg cagttccggc
     2641 ggcggcggcg gcggctacgg gggcggcagc tccggcggcg gcagcagctc cggcggcgga
     2701 tacggcggcg gcagctccag cggaggccac aagtcctcct cttccgggtc cgtgggcgag
     2761 tcttcatcta agggaccaag atactaactc gagtctagag ggcccgttta aacccgctga
     2821 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2881 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2941 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3001 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3061 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3121 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3181 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3241 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3301 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3361 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3541 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3601 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3661 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3721 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3781 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3841 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3901 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3961 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4021 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4081 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4141 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4201 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4261 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4321 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4381 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4441 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4501 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4561 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4621 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4681 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4741 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4801 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4861 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4921 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4981 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5041 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5101 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5161 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5221 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5281 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5341 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5401 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5641 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6001 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6541 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7201 cacatttccc cgaaaagtgc cacctgacgt c
//
