LOCUS       V042716                 8547 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042716
VERSION     V042716
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..3227
                     /label="ADAM8(NM_001109)"
                     /note="ADAM8(NM_001109)"
                     /gene="ADAM8"
     CDS             3234..3299
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3323..3911
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3977..4025
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4227..4655
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4669..4998
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5065..5859
                     /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    6035..6156
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6205..6221)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6229..6245
                     /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(6253..6283)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6298..6319
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6607..7195)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7366..8226)
                     /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(8227..8331)
                     /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 ccaccatgcg cggcctcggg ctctggctgc
      781 tgggcgcgat gatgctgcct gcgattgccc ccagccggcc ctgggccctc atggagcagt
      841 atgaggtcgt gttgccgtgg cgtctgccag gcccccgagt ccgccgagct ctgccctccc
      901 acttgggcct gcacccagag agggtgagct acgtccttgg ggccacaggg cacaacttca
      961 ccctccacct gcggaagaac agggacctgc tgggctccgg ctacacagag acctatacgg
     1021 ctgccaatgg ctccgaggtg acggagcagc ctcgcgggca ggaccactgc ttctaccagg
     1081 gccacgtaga ggggtacccg gactcagccg ccagcctcag cacctgtgcc ggcctcaggg
     1141 gtttcttcca ggtggggtca gacctgcacc tgatcgagcc cctggatgaa ggtggcgagg
     1201 gcggacggca cgccgtgtac caggctgagc acctgctgca gacggccggg acctgcgggg
     1261 tcagcgacga cagcctgggc agcctcctgg gaccccggac ggcagccgtc ttcaggcctc
     1321 ggcccgggga ctctctgcca tcccgagaga cccgctacgt ggagctgtat gtggtcgtgg
     1381 acaatgcaga gttccagatg ctggggagcg aagcagccgt gcgtcatcgg gtgctggagg
     1441 tggtgaatca cgtggacaag ctatatcaga aactcaactt ccgtgtggtc ctggtgggcc
     1501 tggagatttg gaatagtcag gacaggttcc acgtcagccc cgaccccagt gtcacactgg
     1561 agaacctcct gacctggcag gcacggcaac ggacacggcg gcacctgcat gacaacgtac
     1621 agctcatcac gggtgtcgac ttcaccggga ctaccgtggg gtttgccagg gtgtccgcca
     1681 tgtgctccca cagctcaggg gctgtgaacc aggaccacag caagaacccc gtgggcgtgg
     1741 cctgtaccat ggcccatgag atgggccaca acctgggcat ggaccatgat gagaacgtcc
     1801 agggctgccg ctgccaggaa cgcttcgagg ccggccgctg catcatggcg ggcagcattg
     1861 gctccagttt ccccaggatg ttcagtgact gcagccaggc ctacctggag agctttttgg
     1921 agcggccgca gtcggtgtgc ctcgccaacg cccctgacct cagccacctg gtgggcggcc
     1981 ccgtgtgtgg gaacctgttt gtggagcgtg gggagcagtg cgactgcggc ccccccgagg
     2041 actgccggaa ccgctgctgc aactctacca cctgccagct ggctgagggg gcccagtgtg
     2101 cgcacggtac ctgctgccag gagtgcaagg tgaagccggc tggtgagctg tgccgtccca
     2161 agaaggacat gtgtgacctc gaggagttct gtgacggccg gcaccctgag tgcccggaag
     2221 acgccttcca ggagaacggc acgccctgct ccgggggcta ctgctacaac ggggcctgtc
     2281 ccacactggc ccagcagtgc caggccttct gggggccagg tgggcaggct gccgaggagt
     2341 cctgcttctc ctatgacatc ctaccaggct gcaaggccag ccggtacagg gctgacatgt
     2401 gtggcgttct gcagtgcaag ggtgggcagc agcccctggg gcgtgccatc tgcatcgtgg
     2461 atgtgtgcca cgcgctcacc acagaggatg gcactgcgta tgaaccagtg cccgagggca
     2521 cccggtgtgg accagagaag gtttgctgga aaggacgttg ccaggactta cacgtttaca
     2581 gatccagcaa ctgctctgcc cagtgccaca accatggggt gtgcaaccac aagcaggagt
     2641 gccactgcca cgcgggctgg gccccgcccc actgcgcgaa gctgctgact gaggtgcacg
     2701 cagcgtccgg gagcctcccc gtcttcgtgg tggtggttct ggtgctcctg gcagttgtgc
     2761 tggtcaccct ggcaggcatc atcgtctacc gcaaagcccg gagccgcatc ctgagcagga
     2821 acgtggctcc caagaccaca atggggcgct ccaaccccct gttccaccag gctgccagcc
     2881 gcgtgccggc caagggcggg gctccagccc catccagggg cccccaagag ctggtcccca
     2941 ccacccaccc gggccagccc gcccgacacc cggcctcctc ggtggctctg aagaggccgc
     3001 cccctgctcc tccggtcact gtgtccagcc cacccttccc agttcctgtc tacacccggc
     3061 aggcaccaaa gcaggtcatc aagccaacgt tcgcaccccc agtgccccca gtcaaacccg
     3121 gggctggtgc ggccaaccct ggtccagctg agggtgctgt tggcccaaag gttgccctga
     3181 agccccccat ccagaggaag caaggagccg gagctcccac agcacccgga tccgactaca
     3241 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3301 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3361 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3421 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3481 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3541 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3601 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3661 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3721 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3781 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3841 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3901 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3961 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     4021 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4081 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4141 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     4201 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4261 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4321 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4381 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4441 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4501 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4561 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4621 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4681 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4741 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4801 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4861 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4921 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4981 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5041 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5101 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5161 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5221 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5281 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5341 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5401 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5461 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5521 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5581 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5641 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5701 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5761 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5821 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5881 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5941 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6001 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6061 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6121 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6181 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6781 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7021 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8341 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8401 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8461 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8521 gacaattgca tgaagaatct gcttagg
//
