LOCUS       V036107                10467 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036107
VERSION     V036107
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..5147
                     /label="CECR2(NM_001290047)"
                     /note="CECR2(NM_001290047)"
                     /gene="CECR2"
     CDS             5154..5219
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5243..5831
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5897..5945
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6147..6575
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6589..6918
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6985..7779
                     /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    7955..8076
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8125..8141)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8149..8165
                     /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(8173..8203)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8218..8239
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8527..9115)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9286..10146)
                     /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(10147..10251)
                     /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 ccaccatgtg cccagaggag ggcggcgcgg
      781 ccgggctggg cgagctccgc tcctggtggg aggtcccggc catcgcgcac ttctgctcgc
      841 tctttcgcac cgcgttccgc ctgcccgact tcgagatcga ggagttagaa gccgctcttc
      901 acagagatga cgtggagttt atcagtgacc tgattgcctg cctgcttcag ggctgctatc
      961 aacgaagaga tatcacgcct cagacattcc acagctacct agaggacatc atcaactacc
     1021 gctgggagct cgaagaaggg aagcccaacc ctctgaggga agccagtttc caggacctgc
     1081 ctcttcgcac acgggtggag atcctgcacc gactctgtga ttaccggctg gatgcagacg
     1141 atgtcttcga tcttctaaag ggcctggatg cagacagtct ccgtgtggag ccattgggtg
     1201 aagacaattc tggggcacta tattggtatt tctatggaac acgaatgtac aaagaggacc
     1261 cggtgcaagg aaaatccaat ggagaactct ctttgagcag ggaaagtgaa ggacaaaaaa
     1321 atgtctcaag tattcctgga aaaacgggaa aaagaagagg aagaccccca aaacggaaga
     1381 aactgcagga ggagattctg ttgagtgaaa agcaggaaga aaattccttg gcatccgagc
     1441 cacagacaag acatgggtcc caagggccag gccaaggtac ttggtggctc ctgtgccaga
     1501 cagaagagga atggagacag gtcaccgaga gttttcgcga gaggacctcc cttcgagaac
     1561 ggcagctcta caagctcctc agtgaggact tcctgcctga gatctgcaac atgatcgccc
     1621 agaagggaaa acgtccacag cgcacaaagg cagagttgca tcctaggtgg atgtctgacc
     1681 acctgtccat caaacccgtc aagcaagagg agactcctgt gctgaccaga atagaaaaac
     1741 aaaagcgcaa agaggaggaa gaagagcgtc agattcttct agcagtgcag aagaaggagc
     1801 aggagcagat gctaaaggaa gagaggaaac gcgagttgga ggagaaggtc aaggcagtgg
     1861 aagatcgagc gaagaggaga aagctcaggg aagaaagggc atggctgctg gctcaaggaa
     1921 aggagctccc tccagaactt tcccatctgg accccaattc ccccatgaga gaggaaaaaa
     1981 agactaaaga cctctttgag ttggatgatg atttcactgc tatgtataaa gttctagacg
     2041 tggtaaaggc tcacaaggat tcctggccct tcttggaacc tgtggatgaa tcttatgccc
     2101 ctaactatta tcagattatt aaggccccca tggatatttc cagcatggag aagaaactga
     2161 atggaggttt atactgtacc aaggaggaat ttgtaaatga catgaagacc atgttcagga
     2221 attgtcgaaa gtataatggg gaaagtagtg agtataccaa gatgtctgat aatttagaga
     2281 ggtgtttcca tcgggcaatg atgaaacatt ttcctggaga agatggagac acagatgaag
     2341 aattttggat tcgagaggat gaaaagcggg agaaaagacg gagtcgggct gggcgaagtg
     2401 gtgggagcca tgtttggacc cgctccaggg acccagaagg gtccagcagg aaacagcagc
     2461 ccatggagaa tggaggaaag tcgttgcccc ccacacgccg agcgccctct tctggggacg
     2521 atcagagcag cagctccaca cagcccccgc gggaggtggg cacttccaat ggccgaggtt
     2581 tttctcatcc cctgcattgt ggtgggacac ccagccaggc acccttttta aaccagatga
     2641 ggccagcagt accaggaaca tttggccctc tgcgaggatc agatcctgcc accttgtatg
     2701 gctcctctgg agtcccggag ccacaccccg gggagcctgt gcagcagcgt cagcctttca
     2761 ccatgcagcc tccagttgga attaacagcc tccgaggacc caggctaggc acaccagagg
     2821 agaagcaaat gtgcgggggg ctgacacacc tttctaacat gggcccacac cctggatcct
     2881 tgcagcttgg gcagataagt ggcccaagtc aggatggaag catgtatgct ccagctcagt
     2941 tccagccagg attcattcct ccccggcatg ggggggctcc agcccggcca ccagactttc
     3001 ctgaaagctc agaaattcct cccagccata tgtatcgatc gtacaagtac ctgaatcgag
     3061 tacactctgc cgtctggaat gggaaccatg gtgctacgaa ccaaggaccc ttgggcccag
     3121 atgagaagcc ccacctgggg ccaggaccct ctcaccagcc tcgcactctc ggtcacgtga
     3181 tggattcccg agtcatgaga ccacctgtcc cccccaacca gtggactgaa caatcaggct
     3241 tcctacctca tggagttcct tcctcagggt acatgcgacc gccctgcaag tctgccggac
     3301 atcggttaca gccacctcca gtgccagcac ccagttcttt gtttggagca cctgcccagg
     3361 ctcttcgggg ggtgcaggga ggggactcca tgatggacag cccagagatg attgcgatgc
     3421 agcagctctc ctcccgcgtc tgccccccag gtgtgcctta ccacccccac cagcctgcac
     3481 acccccgttt acctggccct tttccgcagg tagctcaccc aatgtcagtc actgtgtcag
     3541 cccccaagcc tgccctgggc aaccctggga gggcaccgga gaacagtgaa gcacaagagc
     3601 ctgagaatga ccaagcagag ccgttgcctg gccttgaaga gaaaccacca ggtgttggta
     3661 cttcagaggg ggtctacctc acacaactac ctcaccccac acctcccctg cagactgact
     3721 gcaccaggca gagctcacca caagaaaggg aaacagtggg cccggagctc aaaagcagct
     3781 cctccgaatc tgcggacaac tgtaaagcaa tgaagggcaa gaatccctgg ccctcggata
     3841 gcagctaccc cggcccagcc gcccaagggt gcgtgagaga cctctccacg gtggcagaca
     3901 ggggcgctct atccgagaac ggagtcattg gggaagcatc tccttgtgga tcggagggga
     3961 agggccttgg tagcagtggt tccgaaaagc tgctctgccc cagaggcaga acgttgcagg
     4021 aaaccatgcc atgcacggga cagaacgcag cgacaccgcc cagcacagac cccggtttga
     4081 cgggaggcac tgtgagccag tttcccccgc tgtatatgcc tggcctagag tacccgaatt
     4141 cagctgccca ttaccacatc agtccaggcc tgcagggtgt gggccctgtg atgggaggga
     4201 agtccccagc atcccatccc cagcattttc ccccaagggg ctttcagtct aaccacccac
     4261 attctggagg ctttccccgg tatcgccccc cacaaggaat gaggtattcc taccacccac
     4321 cgccacagcc ttcctaccac cactatcagc gaactcctta ctatgcctgt ccacagagct
     4381 tttctgactg gcagagacct ctccatcccc agggaagccc aagcggaccc ccagccagtc
     4441 agcctccccc accaaggtcc ctcttctcag ataagaatgc catggccagt ctgcaaggct
     4501 gtgagacact gaatgctgcc ttaacttctc caacccgtat ggatgcagtg gctgctaaag
     4561 tcccaaatga cgggcagaat cctggtccag aggaagagaa gctggatgaa tctatggaga
     4621 ggccagagag tcccaaagaa tttttagacc tggacaacca taacgcagct accaagcggc
     4681 agagctcgtt gtcagccagc gagtatctct atggaactcc tccgcctctg agttcaggaa
     4741 tgggatttgg ttcatctgca tttccacccc acagtgtgat gctgcagacg gggcctccct
     4801 atacccctca gcggccggcc agtcactttc agcccagggc ttactcttcc cctgtggctg
     4861 ccctcccacc tcaccaccca ggggccaccc agcccaacgg cctctctcag gagggtccca
     4921 tctatcgctg ccaggaagaa ggcctgggtc actttcaagc tgtgatgatg gaacaaattg
     4981 gcactagaag tggaataaga ggacctttcc aggaaatgta cagaccatca ggaatgcaga
     5041 tgcacccggt ccagtcgcag gcctcgttcc caaagacccc cacagcagca acatcacagg
     5101 aggaggtgcc gcctcataag cctccaacac ttcccctgga tcagagcgga tccgactaca
     5161 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     5221 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     5281 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5341 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5401 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5461 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5521 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5581 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5641 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5701 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5761 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5821 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     5881 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     5941 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6001 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6061 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     6121 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     6181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6301 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6481 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     6541 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     6601 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6661 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6721 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6841 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6901 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     6961 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     7021 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     7081 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     7141 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     7201 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     7261 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     7321 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     7381 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     7441 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     7501 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     7561 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     7621 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     7681 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     7741 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     7801 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     7861 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     7921 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     7981 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     8041 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     8101 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     8161 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     8221 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     8281 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     8341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     8401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     8461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     8521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     8581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     8641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     8701 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     8761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8941 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     9001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     9061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     9121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     9181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     9241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     9301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     9361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     9421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     9481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     9541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     9601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    10021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    10081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    10141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    10201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    10261 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
    10321 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
    10381 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
    10441 gacaattgca tgaagaatct gcttagg
//
