LOCUS       V034477                 8319 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034477
VERSION     V034477
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..2999
                     /label="ADAM10(NM_001110)"
                     /note="ADAM10(NM_001110)"
                     /gene="ADAM10"
     CDS             3006..3071
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3095..3683
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3749..3797
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3999..4427
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4441..4770
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4837..5631
                     /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    5807..5928
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5977..5993)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6001..6017
                     /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(6025..6055)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6070..6091
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6379..6967)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7138..7998)
                     /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(7999..8103)
                     /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 ccaccatggt gttgctgaga gtgttaattc
      781 tgctcctctc ctgggcggcg gggatgggag gtcagtatgg gaatccttta aataaatata
      841 tcagacatta tgaaggatta tcttacaatg tggattcatt acaccaaaaa caccagcgtg
      901 ccaaaagagc agtctcacat gaagaccaat ttttacgtct agatttccat gcccatggaa
      961 gacatttcaa cctacgaatg aagagggaca cttccctttt cagtgatgaa tttaaagtag
     1021 aaacatcaaa taaagtactc gattatgata cctctcatat ttacactgga catatttatg
     1081 gtgaagaagg aagttttagc catgggtctg ttattgatgg aagatttgaa ggattcatcc
     1141 agactcgtgg tggcacattt tatgttgagc cagcagagag atatattaaa gaccgaactc
     1201 tgccatttca ctctgtcatt tatcatgaag atgatattaa ctatccccat aaatacggtc
     1261 ctcagggggg ctgtgcagat cattcagtat ttgaaagaat gaggaaatac cagatgactg
     1321 gtgtagagga agtaacacag atacctcaag aagaacatgc tgctaatggt ccagaacttc
     1381 tgaggaaaaa acgtacaact tcagctgaaa aaaatacttg tcagctttat attcagactg
     1441 atcatttgtt ctttaaatat tacggaacac gagaagctgt gattgcccag atatccagtc
     1501 atgttaaagc gattgataca atttaccaga ccacagactt ctccggaatc cgtaacatca
     1561 gtttcatggt gaaacgcata agaatcaata caactgctga tgagaaggac cctacaaatc
     1621 ctttccgttt cccaaatatt ggtgtggaga agtttctgga attgaattct gagcagaatc
     1681 atgatgacta ctgtttggcc tatgtcttca cagaccgaga ttttgatgat ggcgtacttg
     1741 gtctggcttg ggttggagca ccttcaggaa gctctggagg aatatgtgaa aaaagtaaac
     1801 tctattcaga tggtaagaag aagtccttaa acactggaat tattactgtt cagaactatg
     1861 ggtctcatgt acctcccaaa gtctctcaca ttacttttgc tcacgaagtt ggacataact
     1921 ttggatcccc acatgattct ggaacagagt gcacaccagg agaatctaag aatttgggtc
     1981 aaaaagaaaa tggcaattac atcatgtatg caagagcaac atctggggac aaacttaaca
     2041 acaataaatt ctcactctgt agtattagaa atataagcca agttcttgag aagaagagaa
     2101 acaactgttt tgttgaatct ggccaaccta tttgtggaaa tggaatggta gaacaaggtg
     2161 aagaatgtga ttgtggctat agtgaccagt gtaaagatga atgctgcttc gatgcaaatc
     2221 aaccagaggg aagaaaatgc aaactgaaac ctgggaaaca gtgcagtcca agtcaaggtc
     2281 cttgttgtac agcacagtgt gcattcaagt caaagtctga gaagtgtcgg gatgattcag
     2341 actgtgcaag ggaaggaata tgtaatggct tcacagctct ctgcccagca tctgacccta
     2401 aaccaaactt cacagactgt aataggcata cacaagtgtg cattaatggg caatgtgcag
     2461 gttctatctg tgagaaatat ggcttagagg agtgtacgtg tgccagttct gatggcaaag
     2521 atgataaaga attatgccat gtatgctgta tgaagaaaat ggacccatca acttgtgcca
     2581 gtacagggtc tgtgcagtgg agtaggcact tcagtggtcg aaccatcacc ctgcaacctg
     2641 gatccccttg caacgatttt agaggttact gtgatgtttt catgcggtgc agattagtag
     2701 atgctgatgg tcctctagct aggcttaaaa aagcaatttt tagtccagag ctctatgaaa
     2761 acattgctga atggattgtg gctcattggt gggcagtatt acttatggga attgctctga
     2821 tcatgctaat ggctggattt attaagatat gcagtgttca tactccaagt agtaatccaa
     2881 agttgcctcc tcctaaacca cttccaggca ctttaaagag gaggagacct ccacagccca
     2941 ttcagcaacc ccagcgtcag cggccccgag agagttatca aatgggacac atgagacgcg
     3001 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3061 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3121 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3181 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3241 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3301 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3361 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3421 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3481 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3541 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3601 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3661 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3721 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3781 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3841 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3901 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3961 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4141 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4441 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4561 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4621 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4681 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4741 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4801 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4861 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4921 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4981 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5041 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5101 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5161 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5221 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5281 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5341 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5401 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5461 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5521 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5581 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5641 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5701 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5761 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5821 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5881 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5941 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6001 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6061 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6181 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6361 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6541 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6781 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8101 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8161 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8221 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8281 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
