LOCUS       V036573                 6813 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036573
VERSION     V036573
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..1493
                     /label="Ywhah(NM_011738)"
                     /note="Ywhah(NM_011738)"
                     /gene="Ywhah"
     CDS             1500..1565
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1589..2177
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2243..2291
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2493..2921
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2935..3264
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3331..4125
                     /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    4301..4422
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4471..4487)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4495..4511
                     /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(4519..4549)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4564..4585
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4873..5461)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5632..6492)
                     /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(6493..6597)
                     /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 ccaccatggg ggatcgagag cagctgctgc
      781 agcgggcgcg actggcggag caggcggagc gctacgacga tatggcctcc gccatgaagg
      841 cggtgacaga gctgaatgaa ccactatcta acgaagatcg aaatctcctc tctgtggcct
      901 acaagaatgt agttggtgcc aggcgatctt cttggagggt tattagtagc attgagcaga
      961 aaaccatggc agatgggaat gagaagaagt tggagaaagt taaagcctac cgggagaaga
     1021 ttgaaaagga gctggagaca gtttgcaatg atgtcttggc tctgcttgac aagttcctta
     1081 tcaagaactg caatgatttt cagtatgaga gcaaggtgtt ctacctgaaa atgaagggcg
     1141 attactaccg ctacctggca gaggtggctt ctggggagaa gaaaaacagt gtggttgaag
     1201 cttctgaggc cgcgtataag gaagccttcg aaatcagcaa agagcacatg cagccaacac
     1261 accccatccg gcttggcctg gccctcaatt tttctgtgtt ctactatgag atccagaatg
     1321 caccagagca agcctgcctc ttagccaaac aagccttcga tgatgctata gctgagctgg
     1381 acacattaaa cgaggattcc tataaggact ccactctcat catgcagttg ctgcgagaca
     1441 acctcaccct ctggacgagc gaccagcagg atgaagaagc aggagaaggc aacggatccg
     1501 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     1561 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     1621 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1681 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1741 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     1801 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     1861 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     1921 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     1981 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2041 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2101 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2161 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     2221 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     2281 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     2341 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     2401 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     2461 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2521 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2581 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2641 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2701 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2761 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2821 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     2881 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2941 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3001 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3061 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3181 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3241 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3301 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3361 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3421 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3481 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3541 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3601 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3661 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3721 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3781 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3841 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3901 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3961 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4021 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4081 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4141 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4201 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4261 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4441 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4501 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4561 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4621 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4681 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4741 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4801 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4861 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4921 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4981 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5041 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5101 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5161 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5221 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5281 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5341 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5401 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5461 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5521 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5581 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5641 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5701 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5761 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5821 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5881 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5941 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6001 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6061 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6121 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6181 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6241 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6301 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6361 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6421 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6481 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6541 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6601 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6661 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     6721 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     6781 ttgaccgaca attgcatgaa gaatctgctt agg
//
