LOCUS       V041490                 8556 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041490
VERSION     V041490
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..3236
                     /label="Adam17(NM_009615)"
                     /note="Adam17(NM_009615)"
                     /gene="Adam17"
     CDS             3243..3308
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3332..3920
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3986..4034
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4236..4664
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4678..5007
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5074..5868
                     /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    6044..6165
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6214..6230)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6238..6254
                     /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(6262..6292)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6307..6328
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6616..7204)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7375..8235)
                     /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(8236..8340)
                     /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 gcggcgtctc ctcatcctga
      781 ccactttggt gcctttcgtc ctggcacccc gacctccgga ggaagcaggc tctggctccc
      841 atccgcgact tgagaagctt gattctttgc tctcagacta cgacatcctc tccttagcta
      901 atattcagca gcactccata aggaaaaggg atctacagtc tgcgacacac ttagaaacat
      961 tactaacttt ttcagctttg aaaagacatt ttaaattata cttgacatca agtaccgaac
     1021 gcttttcaca aaacttgaga gtcgtggtgg tggacgggaa agaagaaagc gagtacagcg
     1081 tgaagtggca gaacttcttc agtggtcacg tggttggtga gcctgactct agggttctag
     1141 cccacatagg agatgatgat gttacagtga gaatcaacac agatggggca gaatataacg
     1201 tagagccact ttggaggttt gtcaatgata ctaaagataa acgaatgctg gtgtataagt
     1261 ctgaagatat caaggatttt tcacgtttgc agtctccaaa agtatgtggt tatttaaatg
     1321 cagatagtga agagctgctt ccaaaagggc tcatagacag agagccatct gaagagtttg
     1381 ttcgtcgagt gaagagacga gctgaaccta accccttgaa gaatacttgt aaattactgg
     1441 tggtagcaga tcatcgattt tataaataca tgggccgtgg agaagagagc accactacaa
     1501 attacttaat agagctaatt gaccgagttg atgacatata ccggaacacg tcgtgggata
     1561 atgcagggtt taaagggtat ggagtgcaga tagagcagat tcgaattctc aagtctccac
     1621 aagaggtaaa acctggtgaa agacacttca atatggcaaa aagtttccca aacgaagaga
     1681 aggatgcttg ggatgtgaag atgctattag agcaatttag ctttgatata gctgaagaag
     1741 catctaaagt ctgcctggct catcttttca cgtaccagga ttttgatatg ggaactcttg
     1801 gattagctta cgttggttct cccagagcaa acagtcatgg aggggtttgt ccaaaagctt
     1861 attacaaccc aactgtgaag aaaaacatct atttaaatag tggtctgact agtactaaaa
     1921 attatggcaa aactattctc acaaaggaag ctgacctggt tacaactcat gaattgggac
     1981 ataattttgg agcagaacat gaccctgatg ggctagcaga atgtgcccca aatgaggacc
     2041 aaggaggaaa gtatgtcatg tatcccatag ctgtgagcgg tgaccacgag aataataaga
     2101 tgttttcaaa ctgcagtaaa cagtccatct acaagaccat agaaagtaag gctcaagagt
     2161 gcttccagga gcgcagcaac aaggtgtgtg gcaactccag ggtggatgaa ggagaggagt
     2221 gtgacccggg tattatgtac ctgaacaacg acacctgctg caatagtgac tgcacactga
     2281 agccgggtgt gcagtgcagt gataggaaca gtccttgctg taaaaactgt cagtttgaga
     2341 cggcgcagaa gaagtgccag gaggctatta atgctacatg caaaggagtg tcttactgca
     2401 cagggaatag cagtgagtgc cccccacccg gagatgctga agatgacact gtgtgcttgg
     2461 accttggcaa gtgcaaggct gggaaatgca tccctttctg caagagggag caggagctgg
     2521 agtcctgcgc atgcgttgac actgacaact cgtgcaaggt gtgctgcagg aacctttctg
     2581 gcccgtgtgt gccgtacgtc gatgcagagc aaaagaactt gtttttgagg aaagggaagc
     2641 catgtacagt agggttttgc gacatgaatg gcaaatgtga gaaacgagta caggacgtaa
     2701 ttgagcgatt ttgggatttc attgaccagc tgagcatcaa cacttttggg aagtttctgg
     2761 cagataacat cgttgggtct gttctggttt tctccttgat attttggatt cctttcagca
     2821 ttcttgtcca ctgtgtggat aagaaactgg acaagcagta tgaatccctg tctctgtttc
     2881 atcacagtaa cattgagatg ctgagcagca tggactcagc atctgttcgc atcatcaagc
     2941 cctttcctgc accccagact ccaggtcgtc tgcaggccct gcagccagct gccatgatgc
     3001 cgccagtacc tgcagctcca aaactggacc accagaggat ggacaccatc caggaagacc
     3061 ccagcacaga ctcacatgca gatgatgacg gttttgagaa ggaccccttc cccaacagca
     3121 gcacagctgc caagtccttt gaggatctca cagaccaccc agtcaccagg agcgaaaagg
     3181 cggcctcatt caagctgcag cgtcagagcc gagttgacag caaagagaca gagtgcggat
     3241 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3301 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     3361 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3421 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3481 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3541 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3601 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3661 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3721 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3781 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3841 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3901 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3961 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     4021 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4081 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4141 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4201 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4261 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4321 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4381 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4441 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4501 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4561 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     4621 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4681 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4741 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4801 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4861 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4921 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4981 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5041 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5101 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5161 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5221 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5281 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5341 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5401 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5461 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5521 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5581 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5641 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5701 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5761 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5821 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5881 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5941 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6001 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6061 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6121 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6181 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6241 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6301 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6361 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6421 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6481 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6781 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7021 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7201 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7321 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7381 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7441 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7501 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7561 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7621 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7681 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7741 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7801 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7861 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7921 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7981 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8041 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8101 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8161 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8221 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8341 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     8401 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8461 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8521 ggcttgaccg acaattgcat gaagaatctg cttagg
//
