LOCUS       V024936                 7347 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024936
VERSION     V024936
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..2027
                     /label="SF3B4(NM_005850)"
                     /note="SF3B4(NM_005850)"
                     /gene="SF3B4"
     CDS             2034..2099
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2123..2711
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2777..2825
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3027..3455
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3469..3798
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3865..4659
                     /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    4835..4956
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5005..5021)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5029..5045
                     /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(5053..5083)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5098..5119
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5407..5995)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6166..7026)
                     /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(7027..7131)
                     /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 tgccgggccg atctccgagc
      781 ggaatcagga tgccactgtg tacgtggggg gcctggatga gaaggttagt gaaccgctgc
      841 tgtgggaact gtttctccag gctggaccag tagtcaacac ccacatgcca aaggatagag
      901 tcactggcca gcaccaaggc tatggctttg tggaattctt gagtgaggaa gatgctgact
      961 atgccattaa gatcatgaac atgatcaaac tctatgggaa gccaatacgg gtgaacaaag
     1021 catcagctca caacaaaaac ctggatgtag gggccaacat tttcattggg aacctggacc
     1081 ctgagattga tgagaagttg ctttatgata ctttcagcgc ctttggggtc atcttacaaa
     1141 cccccaaaat tatgcgggac cctgacacag gcaactccaa aggttatgcc tttattaatt
     1201 ttgcttcatt tgatgcttcg gatgcagcaa ttgaagccat gaatgggcag tacctctgta
     1261 accgtcctat caccgtatct tatgccttca agaaggactc caagggtgag cgccatggct
     1321 cagcagccga acgacttctg gcagctcaga acccgctctc ccaggctgat cgccctcatc
     1381 agctgtttgc agatgcacct cctccaccct ctgctcccaa tcctgtggta tcatcattgg
     1441 ggtctgggct tcctccacca ggcatgcctc ctcctggctc cttcccaccc ccagtgccac
     1501 ctcctggagc cctcccacct gggatacccc cagccatgcc cccaccacct atgcctcctg
     1561 gggctgcagg acatggcccc ccatcggcag gaaccccagg ggcaggacat cctggtcatg
     1621 gacactcaca tcctcaccca ttcccaccgg gtgggatgcc ccatccaggg atgtctcaga
     1681 tgcagcttgc acaccatggc cctcatggct taggacatcc ccacgctgga cccccaggct
     1741 ctgggggcca gccaccgccc cgaccaccac ctggaatgcc tcatcctgga cctcctccaa
     1801 tgggcatgcc cccccgaggg cctccattcg gatctcccat gggtcaccca ggtcctatgc
     1861 ctccgcatgg tatgcgtgga cctcctccac tgatgccccc ccatggatac actggccctc
     1921 cacgaccccc accctatggc taccagcggg ggcctctccc tccacccaga cccactcccc
     1981 ggccaccagt tccccctcga ggcccacttc gaggccctct ccctcaggga tccgactaca
     2041 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2101 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2161 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2221 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2281 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2341 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2401 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2461 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2521 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2581 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2641 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2701 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     2761 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     2821 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     2881 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     2941 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3001 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3061 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3121 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3181 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3241 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3301 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3361 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3421 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3481 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3541 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3601 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3661 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3721 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3781 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3841 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3901 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3961 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4021 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4081 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4141 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4201 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4261 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4321 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4381 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4441 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4501 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4561 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4621 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     4681 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4741 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4801 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4861 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4921 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4981 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5041 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5101 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5221 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5281 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5341 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5401 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5461 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5521 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5581 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5641 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5701 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5761 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5821 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5881 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5941 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6001 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6061 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7141 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7201 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7261 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7321 gacaattgca tgaagaatct gcttagg
//
