LOCUS       V039273                 8367 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039273
VERSION     V039273
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..3047
                     /label="UBTF(NM_014233)"
                     /note="UBTF(NM_014233)"
                     /gene="UBTF"
     CDS             3054..3119
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3143..3731
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3797..3845
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4047..4475
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4489..4818
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4885..5679
                     /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    5855..5976
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6025..6041)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6049..6065
                     /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(6073..6103)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6118..6139
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6427..7015)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7186..8046)
                     /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(8047..8151)
                     /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 ccaccatgaa cggagaagcc gactgcccca
      781 cagacctgga aatggccgcc cccaaaggcc aagaccgttg gtcccaggaa gacatgctga
      841 ctttgctgga atgcatgaag aacaaccttc catccaatga cagctccaag ttcaaaacca
      901 ccgaatcaca catggactgg gaaaaagtag catttaaaga cttttctgga gacatgtgca
      961 agctcaaatg ggtggagatt tctaatgagg tgaggaagtt ccgtacattg acagaattga
     1021 tcctcgatgc tcaggaacat gttaaaaatc cttacaaagg caaaaaactc aagaaacacc
     1081 cagacttccc aaagaagccc ctgacccctt atttccgctt cttcatggag aagcgggcca
     1141 agtatgcgaa actccaccct gagatgagca acctggacct aaccaagatt ctgtccaaga
     1201 aatacaagga gcttccggag aagaagaaga tgaaatatat tcaggacttc cagagagaga
     1261 aacaggagtt cgagcgaaac ctggcccgat tcagggagga tcaccccgac ctaatccaga
     1321 atgccaagaa atcggacatc ccagagaagc ccaaaacccc ccagcagctg tggtacaccc
     1381 acgagaagaa ggtgtatctc aaagtgcggc cagatgccac tacgaaggag gtgaaggact
     1441 ccctggggaa gcagtggtct cagctctcgg acaaaaagag gctgaaatgg attcataagg
     1501 ccctggagca gcggaaggag tacgaggaga tcatgagaga ctatatccag aagcacccag
     1561 agctgaacat cagtgaggag ggtatcacca agtccaccct caccaaggcc gaacgccagc
     1621 tcaaggacaa gtttgacggg cgacccacca agccacctcc gaacagctac tcgctgtact
     1681 gcgcagagct catggccaac atgaaggacg tgcccagcac agagcgcatg gtgctgtgca
     1741 gccagcagtg gaagctgctg tcccagaagg agaaggacgc ctatcacaag aagtgtgatc
     1801 agaaaaagaa agattacgag gtggagctgc tccgtttcct cgagagcctg cctgaggagg
     1861 agcagcagcg ggtcttgggg gaagagaaga tgctgaacat caacaagaag caggccacca
     1921 gccccgcctc caagaagcca gcccaggaag ggggcaaggg cggctccgag aagcccaagc
     1981 ggcccgtgtc ggccatgttc atcttctcgg aggagaaacg gcggcagctg caggaggagc
     2041 ggcctgagct ctccgagagc gagctgaccc gcctgctggc ccgaatgtgg aacgacctgt
     2101 ctgagaagaa gaaggccaag tacaaggccc gagaggcggc gctcaaggct cagtcggaga
     2161 ggaagcccgg cggggagcgc gaggaacggg gcaagctgcc cgagtccccc aaaagagctg
     2221 aggagatctg gcaacagagc gttatcggcg actacctggc ccgcttcaag aatgaccggg
     2281 tgaaggcctt gaaagccatg gaaatgacct ggaataacat ggaaaagaag gagaaactga
     2341 tgtggattaa gaaggcagcc gaagaccaaa agcgatatga gagagagctg agtgagatgc
     2401 gggcacctcc agctgctaca aattcttcca agaagatgaa attccaggga gaacccaaga
     2461 agcctcccat gaacggttac cagaagttct cccaggagct gctgtccaat ggggagctga
     2521 accacctgcc gctgaaggag cgcatggtgg agatcggcag tcgctggcag cgcatctccc
     2581 agagccagaa ggagcactac aaaaagctgg ccgaggagca gcaaaagcag tacaaggtgc
     2641 acctggacct ctgggttaag agcctgtctc cccaggaccg tgcagcatat aaagagtaca
     2701 tctccaataa acgtaagagc atgaccaagc tgcgaggccc aaaccccaaa tccagccgga
     2761 ctactctgca gtccaagtcg gagtccgagg aggatgatga agaggatgag gatgacgagg
     2821 acgaggatga agaagaggaa gatgatgaga atggggactc ctctgaagat ggcggcgact
     2881 cctctgagtc cagcagcgag gacgagagcg aggatgggga tgagaatgaa gaggatgacg
     2941 aggacgaaga cgacgacgag gatgacgatg aggatgaaga taatgagtcc gagggcagca
     3001 gctccagctc ctcctcctca ggggactcct cagactctga ctccaacgga tccgactaca
     3061 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3121 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3181 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3241 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3301 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3361 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3421 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3481 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3541 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3601 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3661 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3721 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3781 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3841 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3901 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3961 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     4021 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4081 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4141 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4201 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4261 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4321 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4381 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4441 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4501 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4561 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4681 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4741 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4801 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4861 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4921 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4981 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5041 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5101 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5161 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5221 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5281 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5341 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5401 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5461 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5521 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5581 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5641 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5701 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5761 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5821 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5941 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6001 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6601 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6841 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8161 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8221 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8281 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8341 gacaattgca tgaagaatct gcttagg
//
