LOCUS       V023607                 7008 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023607
VERSION     V023607
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..1688
                     /label="Hibch(NM_001013112)"
                     /note="Hibch(NM_001013112)"
                     /gene="Hibch"
     CDS             1695..1760
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1784..2372
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2438..2486
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2688..3116
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3130..3459
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3526..4320
                     /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    4496..4617
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4666..4682)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4690..4706
                     /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(4714..4744)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4759..4780
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5068..5656)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5827..6687)
                     /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(6688..6792)
                     /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 ctcggatccg ccaccatggg gcagcagttt gtgtggcggc
      781 tccagtcgag gttcagctca atcagaagag ccagtgttat tctgcaacat ctgagaatgt
      841 ccaagcacac agagactgca gaagtgctgc tggagaggag aggctgtgca ggggtcataa
      901 cgctcaacag acccaagtta ctcaacgctc tgagtctgaa tatgatccgg cagatctatc
      961 cgcagctaaa gaagtgggaa cgagacccag atacattcct gatcatcata aagggagctg
     1021 gaggaaaggc cttctgtgct ggaggggaca tcaaagcact ctcagaagct aaaaaagcag
     1081 ggcagacttt gagtcaagat ttattcagag aagaatatat cctgaacaat gccattgcgt
     1141 cttgtcagaa accgtatgtt gcccttattg atggaatcac aatgggtggg ggagttggtc
     1201 tttcagtcca tggtcaattc cgagtggcta cagaaagatc tctctttgct atgccagaaa
     1261 cgggaatagg actgtttcct gatgtgggcg gaggttattt cttgccaaga cttcaaggaa
     1321 aactgggtta cttccttgca ctgacaggat tcagactaaa aggaagagat gtgcacagag
     1381 cgggaatcgc cacacacttt gtagattcag aaaagttgca tgtactggag gaggagttgt
     1441 tagccttaaa gtccccttca gctgaggatg ttgctggtgt cttagaaagc tatcatgcca
     1501 agtccaagat gggtcaagac aagtctatca tatttgagga acacatggac aaaataaaca
     1561 gttgtttttc agccaatact gtggaacaga ttcttgaaaa cttacggcaa gatggttcac
     1621 cttttgctat ggaacagatt aagttctcat tgacaaagac cagactccaa aatggaaacc
     1681 agctgatcct cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact
     1741 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     1801 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1861 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1921 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     1981 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2041 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2101 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2161 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2221 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2281 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2341 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     2401 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2461 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     2521 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     2581 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     2641 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     2701 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2761 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2821 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     2881 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2941 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3001 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3061 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3121 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3361 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3421 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3481 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3541 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3601 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3661 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3721 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3781 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3841 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3901 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3961 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4021 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4081 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4141 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4201 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4261 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4321 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4381 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4441 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4501 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4561 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4621 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4681 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4741 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4801 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4861 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4921 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4981 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5041 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5101 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5161 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5221 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5281 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5341 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5401 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5461 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5521 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5581 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5641 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5701 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5761 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5821 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5881 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5941 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6001 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6061 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6121 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6181 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6241 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6301 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6361 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6421 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6481 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6541 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6601 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6661 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6721 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6781 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6841 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     6901 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     6961 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
