LOCUS       V028870                 7417 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028870
VERSION     V028870
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     protein_bind    820..838
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    841..859
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     regulatory      1002..1011
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1011..1034
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1038..1061
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1065..3308
                     /label="ADAM10(NM_001110)"
                     /note="ADAM10(NM_001110)"
                     /gene="ADAM10"
     polyA_signal    3375..3599
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    3883..3930
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2μ plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /label="FRT"
     CDS             3938..4960
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     *"
                     /label="HygR"
     polyA_signal    5090..5211
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5260..5276)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5284..5300
                     /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(5308..5338)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5353..5374
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5662..6250)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6421..7281)
                     /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(7282..7386)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cgccaccatg gactacaagg
     1021 acgacgatga caaggcagac tacaaggatg acgatgacaa ggcaatggtg ttgctgagag
     1081 tgttaattct gctcctctcc tgggcggcgg ggatgggagg tcagtatggg aatcctttaa
     1141 ataaatatat cagacattat gaaggattat cttacaatgt ggattcatta caccaaaaac
     1201 accagcgtgc caaaagagca gtctcacatg aagaccaatt tttacgtcta gatttccatg
     1261 cccatggaag acatttcaac ctacgaatga agagggacac ttcccttttc agtgatgaat
     1321 ttaaagtaga aacatcaaat aaagtacttg attatgatac ctctcatatt tacactggac
     1381 atatttatgg tgaagaagga agttttagcc atgggtctgt tattgatgga agatttgaag
     1441 gattcatcca gactcgtggt ggcacatttt atgttgagcc agcagagaga tatattaaag
     1501 accgaactct gccatttcac tctgtcattt atcatgaaga tgatattaac tatccccata
     1561 aatacggtcc tcaggggggc tgtgcagatc attcagtatt tgaaagaatg aggaaatacc
     1621 agatgactgg tgtagaggaa gtaacacaga tacctcaaga agaacatgct gctaatggtc
     1681 cagaacttct gaggaaaaaa cgtacaactt cagctgaaaa aaatacttgt cagctttata
     1741 ttcagactga tcatttgttc tttaaatatt acggaacacg agaagctgtg attgcccaga
     1801 tatccagtca tgttaaagcg attgatacaa tttaccagac cacagacttc tccggaatcc
     1861 gtaacatcag tttcatggtg aaacgcataa gaatcaatac aactgctgat gagaaggacc
     1921 ctacaaatcc tttccgtttc ccaaatattg gtgtggagaa gtttctggaa ttgaattctg
     1981 agcagaatca tgatgactac tgtttggcct atgtcttcac agaccgagat tttgatgatg
     2041 gcgtacttgg tctggcttgg gttggagcac cttcaggaag ctctggagga atatgtgaaa
     2101 aaagtaaact ctattcagat ggtaagaaga agtccttaaa cactggaatt attactgttc
     2161 agaactatgg gtctcatgta cctcccaaag tctctcacat tacttttgct cacgaagttg
     2221 gacataactt tggatcccca catgattctg gaacagagtg cacaccagga gaatctaaga
     2281 atttgggtca aaaagaaaat ggcaattaca tcatgtatgc aagagcaaca tctggggaca
     2341 aacttaacaa caataaattc tcactctgta gtattagaaa tataagccaa gttcttgaga
     2401 agaagagaaa caactgtttt gttgaatctg gccaacctat ttgtggaaat ggaatggtag
     2461 aacaaggtga agaatgtgat tgtggctata gtgaccagtg taaagatgaa tgctgcttcg
     2521 atgcaaatca accagaggga agaaaatgca aactgaaacc tgggaaacag tgcagtccaa
     2581 gtcaaggtcc ttgttgtaca gcacagtgtg cattcaagtc aaagtctgag aagtgtcggg
     2641 atgattcaga ctgtgcaagg gaaggaatat gtaatggctt cacagctctc tgcccagcat
     2701 ctgaccctaa accaaacttc acagactgta ataggcatac acaagtgtgc attaatgggc
     2761 aatgtgcagg ttctatctgt gagaaatatg gcttagagga gtgtacgtgt gccagttctg
     2821 atggcaaaga tgataaagaa ttatgccatg tatgctgtat gaagaaaatg gacccatcaa
     2881 cttgtgccag tacagggtct gtgcagtgga gtaggcactt cagtggtcga accatcaccc
     2941 tgcaacctgg atccccttgc aacgatttta gaggttactg tgatgttttc atgcggtgca
     3001 gattagtaga tgctgatggt cctctagcta ggcttaaaaa agcaattttt agtccagagc
     3061 tctatgaaaa cattgctgaa tggattgtgg ctcattggtg ggcagtatta cttatgggaa
     3121 ttgctctgat catgctaatg gctggattta ttaagatatg cagtgttcat actccaagta
     3181 gtaatccaaa gttgcctcct cctaaaccac ttccaggcac tttaaagagg aggagacctc
     3241 cacagcccat tcagcaaccc cagcgtcagc ggccccgaga gagttatcaa atgggacaca
     3301 tgagacgcta aggggtggga ggtggaaaag gctcgagtct agagggcccg tttaaacccg
     3361 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3421 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3481 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3541 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3601 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3661 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3721 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3781 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3841 cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag
     3901 ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac
     3961 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4021 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4081 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4141 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
     4201 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4261 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
     4321 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4381 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4441 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4501 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4561 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4621 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4681 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
     4741 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4801 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4861 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4921 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga
     4981 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5041 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5101 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5161 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5221 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5281 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5341 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5401 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5461 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5521 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5581 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5641 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5701 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5761 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5821 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5881 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5941 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6001 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6061 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6121 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6181 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6241 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6301 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6361 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6421 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6481 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6541 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6601 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6661 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6721 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6781 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6841 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6901 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6961 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7021 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7081 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7141 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7201 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7261 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7321 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7381 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
