LOCUS       V021430                 9068 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021430
VERSION     V021430
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             683..3715
                     /label="RUBCN(NM_001346873)"
                     /note="RUBCN(NM_001346873)"
                     /gene="RUBCN"
     CDS             3755..3820
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3844..4432
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4498..4546
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4748..5176
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5190..5519
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5586..6380
                     /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    6556..6677
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6726..6742)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6750..6766
                     /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(6774..6804)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6819..6840
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7128..7716)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7887..8747)
                     /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(8748..8852)
                     /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 ggatccgcca ccatgcggcc ggagggcgcg ggaatggagc tcggaggcgg
      721 cgaggagcgc ctgcctgagg agagcaggag ggagcactgg cagttgctgg gtaatttgaa
      781 gacgacggtg gagggtttgg tatcaaccaa cagccccaac gtctggtcta agtatggtgg
      841 cttggagcgg ctttgcaggg acatgcagag catcctctat cacgggctta tccgtgacca
      901 ggcgtgccgc cgccagacgg attactggca gttcgtgaaa gacatccggt ggctcagtcc
      961 ccactcagcc cttcacgtgg agaagttcat cagcgtgcac gagaacgacc agagcagtgc
     1021 tgatggtgcc agtgaacgtg ctgttgccga gctgtggctg cagcacagcc tgcagtacca
     1081 ctgcctctca gcccagctcc ggcccctgct cggggataga cagtatatca gaaaattcta
     1141 cacagatgct gccttcctgc taagtgacgc tcatgtcacg gccatgctgc agtgcctgga
     1201 agcagtggaa cagaacaacc cccgcctcct ggctcagatc gatgcgtcca tgtttgccag
     1261 aaagcacgag agcccgctcc tggtgacaaa gagccagagc ctgacagccc tgcccagttc
     1321 cacatacacc cctccaaaca gctatgctca gcattcctac tttgggtcct tctctagcct
     1381 ccaccaatcc gtgcccaaca atggctcaga gagaagatct acttcctttc cactctctgg
     1441 ccctccccgg aaacctcaag aaagcagagg gcacgtctca ccagcagagg atcaaaccat
     1501 ccaagccccc ccagtttcag tctctgcact agccagggat tcccctttga ccccaaatga
     1561 aatgagctcc agtactctga ccagccccat agaggcatcc tgggtcagca gccagaatga
     1621 ttccccaggt gatgccagtg aggggcctga gtacctggcc attggcaact tggacccccg
     1681 aggccggact gccagctgtc agagtcacag cagcaatgcc gagagcagca gttccaattt
     1741 gttctcctcc agcagctccc agaagccaga ttctgctgcc tcttccttag gggaccagga
     1801 aggaggtggg gagagccagc tgtccagtgt cctccgcagg tccagcttct cagaggggca
     1861 gacactcact gtcaccagtg gggcaaagaa aagccacatt cgctcccatt cggataccag
     1921 cattgcctcc aggggagctc caggaggccc caggaatatc accattatag ttgaagatcc
     1981 cattgcagaa tcctgcaatg ataaggcgaa gttgagaggc cctttgccct actctggtca
     2041 aagcagtgaa gtcagcacac ccagctctct gtacatggaa tatggtggtc ggtacctgtg
     2101 ctcaggggaa ggcatgttcc gaagaccatc agaaggacag tccctcatca gctacctctc
     2161 tgagcaagac ttcggcagct gtgccgacct ggaaaaggag aatgcccact tcagcatctc
     2221 agagtcctta attgctgcca tcgagctaat gaagtgcaac atgatgagcc agtgcctaga
     2281 ggaggaggaa gtggaagagg aagacagtga tagagagatc caggagctga agcagaagat
     2341 ccgccttcgg cgccagcaaa tccgcaccaa gaacctgctc cccatgtacc aggaggctga
     2401 gcacggaagc tttcgggtca cctccagcag ctcccagttc agctcacgtg attcggcaca
     2461 gctctctgac tctggctctg ctgatgaggt tgatgaattt gaaatccaag atggtagcga
     2521 aggatctaac ctgactcaca tatcaaagaa cggactgtca gtgtcactgg cctcgatgtt
     2581 ctcagatgct gacatcagaa ggaacacagc ctcaagcagc aaatccttcg tttcctccca
     2641 gtccttctcc cactgcttcc tgcactccac gtctgctgag gcggtggcca tggggctcct
     2701 gaagcagttt gaggggatgc agcttccagc cgcctcggag ctggagtggc ttgtcccgga
     2761 gcatgatgcc cctcagaagc tcctgcccat tcctgactca ctgcccatct caccggatga
     2821 cgggcagcac gctgacatct acaagctgcg gattcgtgtt cgtggcaact tggagtgggc
     2881 cccgccccgg cctcagataa tttttaatgt tcatccagcc ccaacgagga aaattgccgt
     2941 ggccaagcag aattaccgct gtgcaggatg tggcatccgg actgaccctg attacatcaa
     3001 gcgactgcgg tactgtgagt acctgggcaa gtacttctgc cagtgctgcc acgagaatgc
     3061 ccagatggcc atccccagcc gggttctgcg caagtgggac ttcagcaagt actacgtcag
     3121 caacttctcc aaggacctgc tcattaagat ctggaatgat cctctcttca acgtgcagga
     3181 cataaacagt gccctctata ggaaggtcaa gctgctcaat caagtccggc tgctgcgggt
     3241 ccagctgtgt cacatgaaga acatgttcaa gacttgccga ctggccaagg agcttctgga
     3301 ttcctttgac acagtcccag gccacctgac agaggacctc cacctgtact cactgaatga
     3361 cctgactgcg accaggaagg gggagctggg gccccggctt gctgagctca ccagggcagg
     3421 ggctacccat gtggagagat gcatgctctg ccaagccaaa ggcttcatct gtgagttctg
     3481 tcagaatgag gatgacatca tctttccctt tgagctccat aagtgccgga cctgtgaaga
     3541 gtgtaaagcg tgttaccata aagcctgctt caagtctgga agctgtccgc gctgcgagcg
     3601 gctgcaggcc cggcgggagg cactggccag gcagagcctg gagtcttacc tgtcagacta
     3661 cgaggaggag cccgcggaag cgctggccct ggaagccgcc gtcctggagg ccaccgaatt
     3721 ctgcagatat ccagcacagt ggcggccgct cgaggactac aaagaccatg acggtgatta
     3781 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     3841 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3901 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3961 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4021 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4081 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4141 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4201 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4261 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4321 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4381 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     4441 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4501 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     4561 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4621 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4681 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     4741 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4801 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4861 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     4921 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4981 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5041 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5101 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5161 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5281 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5341 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5401 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5461 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5521 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5581 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5641 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5701 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5761 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5821 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5881 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5941 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6001 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6061 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6121 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6181 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6241 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6301 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6361 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6421 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6481 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6541 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6601 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6661 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6721 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6781 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6841 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6901 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6961 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7021 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7081 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7141 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7201 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7261 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7321 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7381 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7441 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7501 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7561 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7621 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7681 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7741 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7801 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7861 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7921 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7981 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8041 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8101 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8161 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8221 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8281 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8341 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8401 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8461 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8521 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8581 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8641 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8701 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8761 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8821 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8881 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     8941 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     9001 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     9061 tgcttagg
//
