LOCUS       V042235                 7306 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042235
VERSION     V042235
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..1962
                     /label="RNF130(NM_018434)"
                     /note="RNF130(NM_018434)"
                     /gene="RNF130"
     CDS             1969..2058
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2082..2670
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2736..2784
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2986..3414
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3428..3757
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3824..4618
                     /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    4794..4915
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4964..4980)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4988..5004
                     /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(5012..5042)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5057..5078
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5366..5954)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6125..6985)
                     /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(6986..7090)
                     /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 ccaccatgag ctgcgcgggg
      721 cgggcgggcc ctgcccggct cgccgcgctc gccctgctga cctgcagcct gtggccggca
      781 cgggcagaca acgcgagcca ggagtactac acggcgctca tcaacgtgac ggtgcaggag
      841 cccggccgcg gcgccccgct cacgtttcgc atcgaccgcg ggcgctacgg gcttgactcc
      901 cccaaggccg aggtccgcgg ccaggtgctg gcgccgctgc ccctccacgg agttgctgat
      961 catctgggct gtgatccaca aacccggttc tttgtccctc ctaatatcaa acagtggatt
     1021 gccttgctgc agaggggaaa ctgcacgttt aaagagaaaa tatcacgggc cgctttccac
     1081 aatgcagttg ctgtagtcat ctacaataat aaatccaaag aggagccagt taccatgact
     1141 catccaggca ctggagatat tattgctgtc atgataacag aattgagggg taaggatatt
     1201 ttgagttatc tggagaaaaa catctctgta caaatgacaa tagctgttgg aactcgaatg
     1261 ccaccgaaga acttcagccg tggctctcta gtcttcgtgt caatatcctt tattgttttg
     1321 atgattattt cttcagcatg gctcatattc tacttcattc agaagatcag gtacacaaat
     1381 gcacgcgaca ggaaccagcg tcgtctcgga gatgcagcca agaaagccat cagtaaattg
     1441 acaaccagga cagtaaagaa gggtgacaag gaaactgacc cagactttga tcattgtgca
     1501 gtctgcatag agagctataa gcagaatgat gtcgtccgaa ttctcccctg caagcatgtt
     1561 ttccacaaat cctgcgtgga tccctggctt agtgaacatt gtacctgtcc tatgtgcaaa
     1621 cttaatatat tgaaggccct gggaattgtg ccgaatttgc catgtactga taacgtagca
     1681 ttcgatatgg aaaggctcac cagaacccaa gctgttaacc gaagatcagc cctcggcgac
     1741 ctcgccggcg acaactccct tggccttgag ccacttcgaa cttcggggat ctcacctctt
     1801 cctcaggatg gggagctcac tccgagaaca ggagaaatca acattgcagt aacaaaagaa
     1861 tggtttatta ttgccagttt tggcctcctc agtgccctca cactctgcta catgatcatc
     1921 agagccacag ctagcttgaa tgctaatgag gtagaatggt ttctcgagga acaaaaacta
     1981 atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga gcaaaagtta
     2041 atttccgagg aggacttata gtgagggccc gatatctcga caatcaacct ctggattaca
     2101 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2161 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2221 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2281 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2341 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2401 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2461 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     2521 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2581 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2641 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     2701 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     2761 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     2821 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     2881 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     2941 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3001 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3061 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3121 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     3181 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3241 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3301 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     3361 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3421 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3481 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3601 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3661 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3721 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3781 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3841 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3901 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3961 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4021 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4081 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4141 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4201 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4261 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4321 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4381 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4441 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4501 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4561 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4621 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4681 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4741 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4801 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4861 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4921 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4981 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5041 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5101 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5161 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5221 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     5581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6001 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6061 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6121 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6181 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6241 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6301 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6361 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6421 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6481 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6541 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6601 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6661 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6721 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6781 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6841 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6901 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6961 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7021 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7081 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7141 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     7201 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     7261 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
