LOCUS       V028508                 8308 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028508
VERSION     V028508
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             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..1032
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             1039..3828
                     /label="RBM10(NM_005676)"
                     /note="RBM10(NM_005676)"
                     /gene="RBM10"
     polyA_signal    3908..4132
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4178..4606
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4620..4949
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5016..5810
                     /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    5984..6105
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6154..6170)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6178..6194
                     /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(6202..6232)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6247..6268
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6556..7141)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7312..8172)
                     /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(8173..8277)
                     /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 ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaagccta tccctaaccc tctcctcggt
     1021 ctcgattcta caggtaccat ggagtatgaa agacgtggtg gtcgtggtga caggactggc
     1081 cgctatggag ccactgaccg ctcgcaggat gatggtgggg agaaccgcag ccgagaccac
     1141 gactaccggg acatggacta ccgttcatat cctcgcgagt atggcagcca ggagggcaag
     1201 catgactatg acgactcatc tgaggagcag agtgcggagg attcctacga ggcctccccg
     1261 ggctccgaga ctcagcgtag gcggcggcgg cggcacaggc acagccccac cggcccgcca
     1321 ggcttccccc gagacggcga ctatcgggac caggactatc ggaccgagca aggggaggag
     1381 gaggaggagg aggaggatga ggaggaggag gagaaggcca gtaacatcgt catgctgagg
     1441 atgctgccac aggcagccac tgaggatgac atccgtggcc agctgcagtc gcacggcgtg
     1501 caagcacggg aggttcggct gatgcggaac aaatcttcag gtcagagccg gggcttcgcc
     1561 ttcgtcgagt ttagtcactt gcaggacgct acacgatgga tggaagccaa tcagcactcc
     1621 ctcaacatcc tgggccagaa ggtgtcgatg cactacagtg accccaagcc caagatcaat
     1681 gaggactggc tgtgcaataa gtgtggcgtc cagaacttca aacgccgaga gaagtgcttc
     1741 aaatgtggcg tgcccaagtc agaggcagag cagaagctgc ccctcggcac gaggctggat
     1801 cagcagacac tgccactggg tggccgggag ctgagccagg gcctgcttcc cctgccgcag
     1861 ccctaccagg cccagggagt cctggcctcc caagccctgt cacagggctc ggagccaagc
     1921 tcagagaacg ccaatgacac catcattttg cgcaacctga acccacacag caccatggat
     1981 tccatcctgg gggccctggc accctacgcg gtgctgtcct cctccaacgt gcgcgtcata
     2041 aaggacaagc agacccaact gaaccgcggc tttgccttca tccagctctc caccatcgtg
     2101 gaggcagccc agctgctgca gatcctgcag gccctgcacc caccactcac tatcgacggc
     2161 aagaccatca atgttgagtt tgccaagggt tctaagaggg acatggcctc caatgaaggc
     2221 agtcgcatca gtgctgcctc tgtggccagc actgccattg ctgcggccca gtgggccatc
     2281 tcacaggcct cccaaggtgg ggagggtacc tgggccacct ccgaggagcc gccggtcgac
     2341 tacagctact accaacagga tgagggctat ggcaacagcc agggcacaga gtcttccctc
     2401 tatgcccatg gctacctcaa gggcaccaag ggccctggca tcactggaac caaaggggat
     2461 cccactggag caggtcccga ggcctcccta gagcctgggg ccgactctgt gtcgatgcag
     2521 gctttctctc gcgcccagcc tggtgctgct cctggcatct accaacaatc agccgaggcg
     2581 agcagtagcc agggcactgc tgccaacagc cagtcgtata ccatcatgtc acccgctgtg
     2641 ctcaaatctg agctccagag ccctacccat cctagttctg ctctcccacc ggctaccagc
     2701 cccactgccc aggaatccta cagccagtac cctgttcccg acgtctctac ctaccagtac
     2761 gatgagacct ccggctacta ctatgacccc cagaccggcc tctactatga ccccaactcc
     2821 cagtattact acaatgctca gagccagcag tacctgtact gggatgggga gaggcggacc
     2881 tatgttcccg ccctggagca gtcggccgac ggacataagg agacaggggc accctcgaag
     2941 gagggcaaag agaagaagga gaagcacaag accaagacag ctcaacagat tgccaaggac
     3001 atggaacgct gggcccgcag tctcaacaaa caaaaagaaa acttcaaaaa tagcttccag
     3061 cctatcagct ccctgcgaga tgacgagagg cgggagtcag ccactgcaga tgctggctat
     3121 gccatcctcg agaagaaggg agcactagcc gagagacagc acaccagcat ggatctcccg
     3181 aaattggcca gtgacgaccg cccaagccct ccgcgaggac tggtggcagc ctacagcggg
     3241 gagagtgaca gtgaggagga gcaggagcgt gggggccctg agcgggagga gaagctcacc
     3301 gactggcaga agctggcctg tctgctctgc cgacgccagt tccccagcaa agaggcgctc
     3361 atccggcacc agcagctctc agggctccac aagcaaaacc ttgagattca ccggcgagcc
     3421 cacttgtcag aaaacgagct agaagcacta gagaagaatg acatggagca aatgaagtac
     3481 cgggaccgtg cagctgaacg cagagaaaag tatggcatcc ccgagccgcc agagcccaag
     3541 aggaggaagt acggcggcat atccacagcc tctgtagact tcgagcagcc tactcgggac
     3601 gggctgggca gtgacaacat tggcagtcgg atgctgcagg ccatgggctg gaaagagggc
     3661 agcggcctgg gccgcaagaa gcagggcatt gtaacgccta tcgaggccca aacacgggtg
     3721 cggggctccg gcctgggtgc acggggcagc tcctacgggg tcacctcaac cgagtcctac
     3781 aaggagacac tgcacaagac aatggtgacc cgcttcaacg aggcccagtg agaattctgc
     3841 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     3901 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3961 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4021 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4081 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4141 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4201 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4261 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4321 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4381 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4441 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4501 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4561 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4621 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4681 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4741 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4801 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4861 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4921 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4981 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5041 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5101 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5161 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5221 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5281 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5341 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5401 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5461 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5521 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5581 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5641 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5701 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5761 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5821 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5881 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5941 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6001 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6061 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6121 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6181 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6241 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6301 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6361 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6421 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6481 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7081 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7141 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7201 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7261 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7321 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7381 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7441 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7501 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7561 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7621 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7681 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7741 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7801 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7861 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7921 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7981 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8041 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8101 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8161 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8221 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8281 atttccccga aaagtgccac ctgacgtc
//
