LOCUS       V034549                 7368 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034549
VERSION     V034549
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..2048
                     /label="ECSIT(NM_016581)"
                     /note="ECSIT(NM_016581)"
                     /gene="ECSIT"
     CDS             2055..2120
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2144..2732
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2798..2846
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3048..3476
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3490..3819
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3886..4680
                     /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    4856..4977
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5026..5042)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5050..5066
                     /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(5074..5104)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5119..5140
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5428..6016)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6187..7047)
                     /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(7048..7152)
                     /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 ccaccatgag ctgggtccag gccaccctac
      781 tggcccgagg cctctgtagg gcctggggag gcacctgcgg ggccgccctc acaggaacct
      841 ccatctctca ggtccctcgc cggctccctc ggggcctcca ctgcagcgca gctgcccata
      901 gctctgaaca gtccctggtt cccagcccac cggaaccccg gcagaggccc accaaggctc
      961 tggtgccctt tgaggacctg tttgggcagg cgcctggtgg ggaacgggac aaggcgagct
     1021 tcctgcagac ggtgcagaaa tttgcggagc acagcgtgcg taagcggggc cacattgact
     1081 tcatctacct ggccctgcgc aagatgcggg agtatggtgt cgagcgggac ctggctgtgt
     1141 acaaccagct gctcaacatc ttccccaagg aggtcttccg gcctcgcaac atcatccagc
     1201 gcatcttcgt ccactaccct cggcagcagg agtgtgggat tgctgtcctg gagcagatgg
     1261 agaaccacgg tgtgatgccc aacaaggaga cggagttcct gctgattcag atctttggac
     1321 gcaaaagcta ccccatgctc aagttggtgc gcctgaagct gtggttccct cgattcatga
     1381 acgtcaaccc cttcccagtg ccccgggacc tgccccagga ccctgtggag ctggccatgt
     1441 ttggcctgcg gcacatggag cctgacctta gtgccagggt caccatctac caggttcctt
     1501 tgcccaaaga ctcaacaggt gcagcagatc ccccccagcc ccacatcgta ggaatccaga
     1561 gtcccgatca gcaggccgcc ctggcccgcc acaatccagc ccggcctgtc tttgttgagg
     1621 gccccttctc cctgtggctc cgcaacaagt gtgtgtatta ccacatcctc agagctgact
     1681 tgctgccccc ggaggagagg gaagtggaag agacgccgga ggagtggaac ctctactacc
     1741 cgatgcagct ggacctggag tatgtgagga gtggctggga caactacgag tttgacatca
     1801 atgaagtgga ggaaggccct gtcttcgcca tgtgcatggc gggtgctcat gaccaggcga
     1861 cgatggctaa gtggatccag ggcctgcagg agaccaaccc aaccctggcc cagatccccg
     1921 tggtcttccg cctcgccggg tccacccggg agctccagac atcctctgca gggctggagg
     1981 agccgcccct gcccgaggac caccaggaag aagacgacaa cctgcagcga cagcagcagg
     2041 gccagagcgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2101 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2161 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2221 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2281 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2341 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2401 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2461 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2521 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2581 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2641 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2701 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     2761 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2821 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     2881 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     2941 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3001 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3061 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3121 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3181 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3241 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3301 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3361 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3421 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3481 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3541 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3661 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3721 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3781 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3841 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3901 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3961 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4021 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4081 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4141 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4201 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4261 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4321 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4381 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4441 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4501 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4561 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4621 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4681 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4741 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4801 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4981 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5041 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5101 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5161 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5221 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5281 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5341 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5401 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5461 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5521 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5581 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5641 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5701 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5761 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5821 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5881 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5941 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7201 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7261 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7321 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
