LOCUS       V018889                 7526 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018889
VERSION     V018889
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"
     CDS             683..2206
                     /label="GSDMC(NM_031415)"
                     /note="GSDMC(NM_031415)"
                     /gene="GSDMC"
     CDS             2213..2278
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2302..2890
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2956..3004
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3206..3634
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3648..3977
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4044..4838
                     /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    5014..5135
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5184..5200)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5208..5224
                     /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(5232..5262)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5277..5298
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5586..6174)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6345..7205)
                     /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(7206..7310)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgccctc catgttggaa cgcattagca aaaatttggt
      721 caaagagatt ggaagcaaag acctgacacc tgtcaaatac ctattgagtg ccaccaaatt
      781 acgtcagttt gttatattac gaaagaagaa ggattctcgt tcatcatttt gggaacaatc
      841 tgactatgtt ccagttgaat tctccctcaa tgacatcctg gagccaagtt cttcagtcct
      901 agaaactgtt gtgacaggac cgttccactt cagtgacatt atgatccaga agcataaggc
      961 tgacatgggt gtgaatgttg gtatagaagt gagtgtgtca ggggaggcct ctgtggacca
     1021 tggatgctcc ctcgagtttc aaattgttac catcccatca ccaaacctgg aagactttca
     1081 aaaaaggaaa ctgttggatc cagagccatc atttctgaag gagtgccgga ggagagggga
     1141 caacctgtac gtggtgacag aggctgttga actgatcaac aatactgtgc tgtacgatag
     1201 cagtagtgtg aatattttag ggaaaattgc tctttggatt acctatggca agggtcaagg
     1261 ccaaggagag agtctcagag tgaagaagaa ggcgctgact cttcagaaag gcatggtgat
     1321 ggcttataag agaaagcagc tggttatcaa ggagaaagcc attctcatct cagatgatga
     1381 tgaacagaga acctttcaag atgagtacga aatttccgaa atggtaggct actgtgctgc
     1441 gaggagtgag gggttgctac catcatttca taccatctct ccaaccctct tcaatgcctc
     1501 atccaatgat atgaagttaa aaccagagct atttctgaca cagcaatttt tgagcgggca
     1561 tttgccaaaa tacgaacaag ttcacatcct cccagtagga agaatagagg aacccttctg
     1621 gcaaaatttc aagcatctac aagaggaggt tttccagaaa ataaagacac tggctcagct
     1681 ctcaaaggat gttcaggatg tcatgttcta cagtatcctg gccatgctca gagacagagg
     1741 ggctctacag gacctgatga acatgctgga attggacagc tcaggtcatt tggatggccc
     1801 tggtggtgcc atcctaaaga aacttcaaca ggattcaaac catgcatggt ttaacccaaa
     1861 ggaccccatt ctttatctcc ttgaagccat aatggtgctg agtgacttcc aacacgattt
     1921 gctggcctgt tccatggaga agaggatcct gcttcagcaa caggagctgg taaggagcat
     1981 cctggagcca aacttcagat acccctggag cattcccttc accctcaaac ctgagctcct
     2041 cgccccactc cagagtgagg gtttggccat cacctatggc ctgctggagg agtgtggcct
     2101 taggatggag ctggataacc ccaggtcaac ctgggatgta gaagcaaaga tgcccctgtc
     2161 tgccctctat gggactctct cgttgctgca gcagctggct gaggccctcg aggactacaa
     2221 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     2281 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     2341 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2401 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2461 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2521 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2581 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2641 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     2701 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2761 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2821 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2881 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     2941 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     3001 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     3061 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     3121 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     3181 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3901 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3961 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4021 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4081 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4141 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4201 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4261 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4321 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4381 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4441 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4501 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4561 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4621 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4681 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4741 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4801 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     4861 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4921 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4981 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5161 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5221 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5281 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5341 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5761 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6001 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6481 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7321 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     7381 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     7441 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     7501 acaattgcat gaagaatctg cttagg
//
