LOCUS       V020317                 7217 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020317
VERSION     V020317
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..1897
                     /label="Rbm17(NM_152824)"
                     /note="Rbm17(NM_152824)"
                     /gene="Rbm17"
     CDS             1904..1969
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1993..2581
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2647..2695
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2897..3325
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3339..3668
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3735..4529
                     /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    4705..4826
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4875..4891)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4899..4915
                     /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(4923..4953)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4968..4989
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5277..5865)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6036..6896)
                     /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(6897..7001)
                     /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 ccatgtccct atatgatgac ctgggagtgg agaccagtga
      721 ctcaaaaact gaaggctggt ccaaaaactt caagctcctg cagtcccagc tccaggtgaa
      781 gaaggcggcg ctcactcagg ccaagagcca aaggaccaag caaagtacag tgcttgctcc
      841 ggtcatcgac ctaaagcgag gcggctcctc agatgaccgg cagattgcag acacaccacc
      901 tcacgtggca gctgggctga aggaccctgt gcccagtggg ttttctgcag gggaagttct
      961 gatcccctta gctgatgaat atgaccctat gttccccaat gactatgaga aagtggtgaa
     1021 gcgccagaga gaagagcggc agaggcagcg ggagctggaa agacagaagg aaatagagga
     1081 aagagaaaag aggcgtaaag acagacacga agccagtggg ttttcaagac gaccagaccc
     1141 tgattctgat gaggatgaag attatgagcg agagcggagg aaaagaagta tgggaggagc
     1201 tgccatcgcc ccaccgacgt ctcttgtaga gaaagacaaa gagttacccc gcgattttcc
     1261 ttatgaagag gactcaagac cgagatcaca gtcttccaaa gctgctattc ctccccccgt
     1321 gtatgaggag ccggacagac caagatctcc aacaggcccc agcaactcct tccttgctaa
     1381 catgggtggc acagtggctc ataagattat gcagaagtat ggcttccggg aaggtcaggg
     1441 actggggaaa cacgagcaag ggctgagtac tgcattgtct gtggagaaga ccagcaagcg
     1501 tggcggcaag atcattgtgg gggatgcgac agagaagggc gaggctcagg atgcatccaa
     1561 gaagtcggat tcaaatccat taactgaaat tcttaagtgc cctactaaag tggtcttgct
     1621 gaggaacatg gttggtgcag gagaggtcga tgaagacttg gaagttgaaa ccaaggaaga
     1681 atgtgaaaaa tatggcaaag ttgggaaatg tgtgatattt gagattcctg gtgcccctga
     1741 tgatgaagca gtacggatat ttttagaatt tgagagagtc gaatcagcaa ttaaagctgt
     1801 ggtggatctg aatgggaggt attttggtgg acgggtggta aaagcatgtt tctacaattt
     1861 ggataaattc agggtcttgg atctagcaga gcaagttctc gaggactaca aagaccatga
     1921 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     1981 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2041 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2101 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2161 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2221 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2281 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2341 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     2401 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2461 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2521 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2581 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     2641 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     2701 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     2761 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     2821 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     2881 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3241 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3301 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3361 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3421 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3541 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3601 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3661 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3721 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3781 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3841 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3901 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3961 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4021 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4081 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4141 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4201 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4261 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4321 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4381 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4441 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4501 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     4561 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4621 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4681 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4741 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4801 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4861 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4921 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4981 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5041 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5101 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5161 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5221 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5281 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5341 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5401 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5461 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5521 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5581 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5641 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5701 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5761 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5821 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5881 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5941 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6001 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6061 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7021 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     7081 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7141 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     7201 tgaagaatct gcttagg
//
