LOCUS       V024014                 8894 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024014
VERSION     V024014
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             779..3574
                     /label="RBM12(NM_001198840)"
                     /note="RBM12(NM_001198840)"
                     /gene="RBM12"
     CDS             3581..3646
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3670..4258
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4324..4372
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4574..5002
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5016..5345
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5412..6206
                     /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    6382..6503
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6552..6568)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6576..6592
                     /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(6600..6630)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6645..6666
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6954..7542)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7713..8573)
                     /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(8574..8678)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggctgtggtc atccgtttgc aaggtctccc aattgtggcg gggaccatgg acattcgcca
      841 cttcttctct ggattgacca ttcctgatgg gggcgtgcat attgtagggg gtgaactggg
      901 tgaggctttc atcgtttttg ccactgatga agatgcaagg cttggtatga tgcgcacagg
      961 tggtacaatt aaagggtcaa aagtaacact attgttgagt agtaagacgg aaatgcagaa
     1021 tatgattgaa ctgagtcgta ggcgttttga aactgccaac ttagatatac caccagcaaa
     1081 tgccagtaga tcaggaccac cacctagctc aggaatgagt agcagggtaa acttgcccac
     1141 aacagtatcc aactttaata atccatcacc cagtgtagtt actgccacca cttctgttca
     1201 tgaaagcaac aaaaacatac agacattttc cacagccagc gtaggaacag ctcctccaaa
     1261 tatgggggct tcctttggga gcccaacgtt tagctcaact gttccaagca cagcctctcc
     1321 aatgaacaca gtcccgccgc caccaattcc tccaattcca gcgatgccat ctctgccacc
     1381 aatgccatcc attcccccaa ttccagttcc tcctccagta cctacattgc ctcctgtgcc
     1441 tcctgtgccc ccgattcccc cagttccttc tgtgccaccc atgaccccac tgccacccat
     1501 gtcgggcatg ccgcccttga atccgccacc tgtggcacct ctacctgctg gaatgaatgg
     1561 ctctggagca cctatgaatt tgaacaataa tctgaatcct atgtttcttg gtccgttgaa
     1621 tcctgttaac cctatccaga tgaactctca gagcagtgtg aagccactcc ccatcaaccc
     1681 tgatgatctg tatgtcagtg tgcatggaat gcccttttct gcaatggaaa atgatgtcag
     1741 agattttttt catgggctcc gtgttgatgc agtgcatttg ttgaaagatc atgtaggtcg
     1801 aaataatggg aatggattgg ttaagtttct ctcccctcaa gatacatttg aagctttgaa
     1861 acgaaacaga atgctgatga ttcaacgcta tgtggaagtt agccctgcca cagaaagaca
     1921 gtgggtagct gctggaggcc atatcacttt taagcaaaat atgggacctt ctggacaaac
     1981 tcatccccct cctcagacac ttcccaggtc aaaatcgccc agtgggcaga aaagatcaag
     2041 gtcaagatca ccacatgagg ctggtttttg tgtttacttg aaagggctac catttgaagc
     2101 agaaaacaaa catgtcattg atttttttaa aaagctggat attgtggaag atagtattta
     2161 tatagcttat ggacccaatg ggaaagcaac tggcgaaggc tttgtagagt tcagaaatga
     2221 ggctgactat aaggctgctc tgtgtcgtca taaacagtac atgggcaatc gctttattca
     2281 agttcatcca attactaaga aaggtatgct agaaaagata gatatgattc gaaaaagact
     2341 gcagaacttc agctatgacc agagggaaat gatactaaat ccagaggggg atgtcaactc
     2401 tgccaaagtc tgtgcccaca taacaaatat tccattcagc attacaaaga tggatgttct
     2461 tcagttccta gaaggaatcc cagtggatga aaatgctgta catgttcttg ttgataacaa
     2521 tgggcaaggt ctaggacagg cattggttca gtttaaaaat gaagatgatg cacgtaagtc
     2581 tgaacgctta caccgtaaaa aacttaatgg gagagaagct tttgttcatg tagttaccct
     2641 agaagatatg agagagattg agaaaaatcc ccctgcccaa ggaaaaaagg gattaaagat
     2701 gcctgtgcca ggtaatcctg cagttccagg aatgcccaat gcgggactgc ccggtgtggg
     2761 actgcccagt gcaggacttc ccggtgcagg cctgcccagc acaggactgc ctggttcagc
     2821 aataaccagt gcaggactgc ctggtgcggg aatgcccagt gcaggaatac ctagtgcagg
     2881 aggtgaagag catgccttcc tgactgtagg atcaaaggaa gccaataatg ggcctccatt
     2941 taactttcct ggtaattttg gtggatcaaa tgcctttggg ccaccaatcc ctcctccagg
     3001 attaggaggc ggggcctttg gtgatgctag gcctggtatg ccttcagttg gaaacagtgg
     3061 tttgcctggt ctaggactgg atgttccggg ttttggaggt ggaccaaaca atttaagtgg
     3121 gccatcggga tttggagggg gccctcagaa ttttggaaat ggccctggta gcttaggcgg
     3181 tcccccgggg tttggaagtg gccctcctgg tcttggaagt gcccctgggc atttgggtgg
     3241 gccaccagct tttgggcctg gccccggccc cggccccggc cctggcccaa tccatattgg
     3301 tggtccccct ggctttgcat ctagttctgg aaaaccagga ccgacagtaa ttaaagtgca
     3361 aaacatgccc tttactgtgt ctattgatga gattttagat ttcttttatg gctatcaagt
     3421 aatcccaggc tcagtgtgtt taaaatacaa tgaaaaaggt atgcccacag gtgaagccat
     3481 ggtggccttt gagtctcggg atgaagccac agctgctgtc attgacttaa atgacaggcc
     3541 tataggttca agaaaagtaa aacttgtatt agggctcgag gactacaaag accatgacgg
     3601 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     3661 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3721 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3781 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3841 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3901 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3961 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4021 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     4081 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4141 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4201 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4261 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4321 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     4381 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4441 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4501 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     4561 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4621 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4681 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     4741 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4801 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4861 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4921 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     4981 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5041 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5101 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5161 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5221 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5281 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5341 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5401 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5461 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5521 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5581 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5641 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5701 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5761 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5821 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5881 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5941 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     6001 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     6061 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6121 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6181 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     6241 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6301 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6361 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     6421 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6481 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     6541 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6601 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6661 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6721 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6781 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6841 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6901 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6961 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7021 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7081 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7141 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7201 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7261 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7321 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7381 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7441 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7501 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7561 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7621 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7681 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7741 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7801 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7861 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7921 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7981 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8041 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8101 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8161 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8221 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8281 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8341 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8401 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8461 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8521 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8581 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8641 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8701 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     8761 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8821 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8881 agaatctgct tagg
//
