LOCUS       V027915                 7389 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027915
VERSION     V027915
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2069
                     /label="RNF150(NM_020724)"
                     /note="RNF150(NM_020724)"
                     /gene="RNF150"
     CDS             2076..2141
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2165..2753
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2819..2867
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3069..3497
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3511..3840
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3907..4701
                     /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    4877..4998
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5047..5063)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5071..5087
                     /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(5095..5125)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5140..5161
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5449..6037)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6208..7068)
                     /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(7069..7173)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc aatgtctctc atccaagcgt
      781 gctgcagtct ggctctctca acatggctgc tttccttttg tttcgtgcat ctgctctgcc
      841 tggactttac cgtggccgag aaggaggaat ggtacaccgc cttcgtgaac atcacctacg
      901 ccgagcccgc gccggacccc ggggccgggg cggcgggcgg cggcggcgcg gagctgcaca
      961 cggagaagac ggagtgcggg cgctacggag agcactcgcc caagcaggac gcccgcgggg
     1021 aggtggtcat ggccagctcg gcccacgacc gcctggcctg cgaccccaac accaagttcg
     1081 ccgccccgac ccgcggcaag aactggatag ccctcatccc caagggcaac tgcacgtaca
     1141 gggataagat ccggaacgcg ttcctgcaga acgcctcagc cgtggtcatc ttcaacgtgg
     1201 gctccaacac caacgagacc atcaccatgc cccacgcggg tgtagaagac atcgtggcca
     1261 taatgattcc tgagccaaaa gggaaggaga tagtaagcct gctggaaaga aacatcaccg
     1321 tgacaatgta catcaccatc ggaacccgga acttgcagaa atatgtgagc cgcacttcgg
     1381 ttgtgtttgt ctccatctcc ttcattgtcc tgatgatcat ttccctcgca tggctcgtct
     1441 tttattacat ccagaggttt cgatatgcaa atgccaggga taggaaccag cgccgactgg
     1501 gggatgcagc aaagaaagcc atcagcaaac tccagatcag gaccatcaag aagggtgaca
     1561 aggaaacaga gtctgatttt gacaactgtg cagtttgtat tgaagggtac aagcccaatg
     1621 acgttgtccg gatcctgccc tgccggcatc ttttccacaa gtcctgtgtt gacccctggc
     1681 ttctagacca tcgtacctgt cccatgtgca agatgaacat tcttaaagcc ctagggatcc
     1741 cgcccaatgc cgactgcatg gacgacttgc ccactgactt cgagggctct ctgggaggtc
     1801 cacccaccaa ccagatcaca ggtgccagcg acacaacagt gaatgaaagt tcagtcactt
     1861 tggaccctgc tgtccggact gtgggagcct tgcaggtggt ccaggataca gaccccatcc
     1921 cccaggaggg agacgtcatc tttactacta acagtgagca ggagccagct gtaagcagtg
     1981 attctgacat ttccttgatc atggcaatgg aggttggact gtctgatgta gaactttcca
     2041 ctgaccagga ctgtgaagaa gtgaaatctg gatccgacta caaagaccat gacggtgatt
     2101 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     2161 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2221 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2281 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2341 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2401 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2461 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2521 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2581 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2641 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2701 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2761 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2821 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     2881 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     2941 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3001 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3061 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3421 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3481 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3661 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3721 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3781 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3841 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4081 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4681 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4741 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4801 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4861 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4921 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4981 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5041 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5101 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5161 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5221 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5281 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5341 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5641 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5881 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6181 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6241 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6301 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6361 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6421 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6481 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6541 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6601 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6661 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6721 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6781 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6841 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6901 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6961 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7021 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7081 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7141 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7201 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7261 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7321 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7381 ctgcttagg
//
