LOCUS       V047357                 7827 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047357
VERSION     V047357
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..2507
                     /label="IRF2BP1(NM_015649)"
                     /note="IRF2BP1(NM_015649)"
                     /gene="IRF2BP1"
     CDS             2514..2579
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2603..3191
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3257..3305
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3507..3935
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3949..4278
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4345..5139
                     /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    5315..5436
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5485..5501)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5509..5525
                     /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(5533..5563)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5578..5599
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5887..6475)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6646..7506)
                     /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(7507..7611)
                     /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 gtctgtgcag gcgtcccgcc
      781 gccagtggtg ctacctgtgc gacctgccca agatgccgtg ggccatggtg tgggacttca
      841 gcgaggccgt gtgtcgcggc tgcgtgaact tcgagggcgc ggaccgcatc gaactgctca
      901 tcgatgccgc ccgccagctc aagcgcagcc acgtgctccc cgagggccgc tcgcccgggc
      961 ccccggccct taagcacccg gccaccaagg acctggcggc ggcagccgca caggggcccc
     1021 agctgccgcc cccgcaggcc cagccccagc cgtcagggac cggcggcggc gtgtcgggcc
     1081 aggaccgcta tgacagggcc acatcatcag gccgcctccc cctgccctcg cccgccctgg
     1141 agtacactct ggggtcccgc ctggccaatg ggctgggccg tgaggaggcc gtggctgagg
     1201 gggcgcgaag ggccttgctt ggctccatgc ctggcttgat gccccctggg ctgctggcag
     1261 ctgcagtgtc tggcctggga agccgaggcc tgacgctggc acccggcttg agtcctgccc
     1321 gtcccctctt cggctccgat ttcgagaaag agaagcagca gaggaatgcg gactgtctgg
     1381 cagaactgaa cgaggccatg cgaggccggg cagaggaatg gcacgggcgc cccaaagcag
     1441 tgcgggaaca gctactggcg ctgtccgcct gcgccccgtt caatgtgcgc ttcaagaagg
     1501 atcacgggct ggtggggcga gtgttcgcct tcgatgctac tgcccgtcct ccaggatacg
     1561 agttcgagct gaagctcttc accgaatacc cctgtggttc cggcaatgtg tacgccggcg
     1621 tcctggcagt ggctcgccag atgttccacg atgctctgcg ggagccgggc aaggcactgg
     1681 cttcttcggg cttcaagtac ctcgaatatg aacgccggca tggatcagga gaatggcggc
     1741 agctgggcga gctgcttacc gacggcgtcc gcagcttccg cgagccagct cccgcggagg
     1801 ccctgcccca gcagtaccca gagccggccc ctgcggctct ctgtggccca cccccgcgag
     1861 ccccatcccg gaacctggcg cccacgccgc gccgtcgcaa ggcatccccc gagccggagg
     1921 gcgaggcggc tgggaagatg accaccgagg agcagcagca acggcactgg gtggcacccg
     1981 gcggcccgta ctccgctgag acccctggtg tgccctcgcc cattgccgcc ctgaagaatg
     2041 tggccgaagc cctgggccac tcacccaagg accctggcgg aggcgggggg cctgtgcgtg
     2101 cagggggcgc cagccctgca gcctcctcca cggcccagcc gccaacccag catcgccttg
     2161 tggcccgcaa cggcgaagca gaagtcagtc ccacagcggg ggccgaagct gtcagcgggg
     2221 gtggcagcgg cactggggcg acccctgggg cccccctgtg ctgtaccctg tgcagggagc
     2281 ggctagaaga cacccacttc gtccagtgcc cctcggtgcc cggacacaag ttctgctttc
     2341 cctgctcccg ggagttcatc aaggcgcagg gcccggccgg ggaggtgtac tgcccgagcg
     2401 gagacaagtg cccgctggtc ggctcctccg tgccctgggc cttcatgcag ggcgagatcg
     2461 ccaccatcct tgctggagac atcaaagtta agaaagaacg ggaccccgga tccgactaca
     2521 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2581 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2641 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2701 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2761 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2821 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2881 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2941 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3001 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3061 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3121 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3181 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3241 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3301 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3361 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3421 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3481 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3541 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3601 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3661 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3721 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3781 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3841 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3901 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4321 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4381 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4441 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4501 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4561 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4621 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4681 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4741 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4801 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4861 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4921 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4981 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5041 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5101 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5161 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5221 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5281 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5341 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5401 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5461 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5521 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5581 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5641 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5701 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5761 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5821 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5881 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5941 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6001 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6061 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6121 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6181 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6241 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6301 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6361 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6421 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6481 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6541 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6601 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6661 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6721 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6781 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6841 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6901 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6961 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7021 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7081 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7141 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7201 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7261 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7321 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7381 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7441 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7501 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7561 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7621 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7681 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7741 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7801 gacaattgca tgaagaatct gcttagg
//
