LOCUS       V027806                 7764 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027806
VERSION     V027806
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..3653
                     /label="ADAM15(NM_207197)"
                     /note="ADAM15(NM_207197)"
                     /gene="ADAM15"
     polyA_signal    3722..3946
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4230..4277
                     /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             4285..5307
                     /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    5437..5558
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5607..5623)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5631..5647
                     /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(5655..5685)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5700..5721
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6009..6597)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6768..7628)
                     /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(7629..7733)
                     /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 ggcaatgcgg ctggcgctgc
     1081 tctgggccct ggggctcctg ggcgcgggca gccctctgcc ttcctggccg ctcccaaata
     1141 taggtggcac tgaggagcag caggcagagt cagagaaggc cccgagggag cccttggagc
     1201 cccaggtcct tcaggacgat ctcccaatta gcctcaaaaa ggtgcttcag accagtctgc
     1261 ctgagcccct gaggatcaag ttggagctgg acggtgacag tcatatcctg gagctgctac
     1321 agaataggga gttggtccca ggccgcccaa ccctggtgtg gtaccagccc gatggcactc
     1381 gggtggtcag tgagggacac actttggaga actgctgcta ccagggaaga gtgcggggat
     1441 atgcaggctc ctgggtgtcc atctgcacct gctctgggct cagaggcttg gtggtcctga
     1501 ccccagagag aagctatacc ctggagcagg ggcctgggga ccttcagggt cctcccatta
     1561 tttcgcgaat ccaagatctc cacctgccag gccacacctg tgccctgagc tggcgggaat
     1621 ctgtacacac tcagaagcca ccagagcacc ccctgggaca gcgccacatt cgccggaggc
     1681 gggatgtggt aacagagacc aagactgtgg agttggtgat tgtggctgat cactcggagg
     1741 cccagaaata ccgggacttc cagcacctgc taaaccgcac actggaagtg gccctcttgc
     1801 tggacacatt cttccggccc ctgaatgtac gagtggcact agtgggcctg gaggcctgga
     1861 cccagcgtga cctggtggag atcagcccaa acccagctgt caccctcgaa aacttcctcc
     1921 actggcgcag ggcacatttg ctgcctcgat tgccccatga cagtgcccag ctggtgactg
     1981 gtacttcatt ctctgggcct acggtgggca tggccattca gaactccatc tgttctcctg
     2041 acttctcagg aggtgtgaac atggaccact ccaccagcat cctgggagtc gcctcctcca
     2101 tagcccatga gttgggccac agcctgggcc tggaccatga tttgcctggg aatagctgcc
     2161 cctgtccagg tccagcccca gccaagacct gcatcatgga ggcctccaca gacttcctac
     2221 caggcctgaa cttcagcaac tgcagccgac gggccctgga gaaagccctc ctggatggaa
     2281 tgggcagctg cctcttcgaa cggctgccta gcctaccccc tatggctgct ttctgcggaa
     2341 atatgtttgt ggagccgggc gagcagtgtg actgtggctt cctggatgac tgcgtcgatc
     2401 cctgctgtga ttctttgacc tgccagctga ggccaggtgc acagtgtgca tctgacggac
     2461 cctgttgtca aaattgccag ctgcgcccgt ctggctggca gtgtcgtcct accagagggg
     2521 attgtgactt gcctgaattc tgcccaggag acagctccca gtgtccccct gatgtcagcc
     2581 taggggatgg cgagccctgc gctggcgggc aagctgtgtg catgcacggg cgttgtgcct
     2641 cctatgccca gcagtgccag tcactttggg gacctggagc ccagcccgct gcgccacttt
     2701 gcctccagac agctaatact cggggaaatg cttttgggag ctgtgggcgc aaccccagtg
     2761 gcagttatgt gtcctgcacc cctagagatg ccatttgtgg gcagctccag tgccagacag
     2821 gtaggaccca gcctctgctg ggctccatcc gggatctact ctgggagaca atagatgtga
     2881 atgggactga gctgaactgc agctgggtgc acctggacct gggcagtgat gtggcccagc
     2941 ccctcctgac tctgcctggc acagcctgtg gccctggcct ggtgtgtata gaccatcgat
     3001 gccagcgtgt ggatctcctg ggggcacagg aatgtcgaag caaatgccat ggacatgggg
     3061 tctgtgacag caacaggcac tgctactgtg aggagggctg ggcaccccct gactgcacca
     3121 ctcagctcaa agcaaccagc tccctgacca cagggctgct cctcagcctc ctggtcttat
     3181 tggtcctggt gatgcttggt gccagctact ggtaccgtgc ccgcctgcac cagcgactct
     3241 gccagctcaa gggacccacc tgccagtaca gggcagccca atctggtccc tctgaacggc
     3301 caggacctcc gcagagggcc ctgctggcac gaggcactaa gcaggctagt gctctcagct
     3361 tcccggcccc cccttccagg ccgctgccgc ctgaccctgt gtccaagaga ctccaggctg
     3421 agctggctga ccgacccaat ccccctaccc gccctctgcc cgctgacccg gtggtgagaa
     3481 gcccgaagtc tcaggggcca gccaagcccc cacccccaag gaagccactg cctgccgacc
     3541 cccagggccg gtgcccatcg ggtgacctgc ccggcccagg ggctggaatc ccgcccctag
     3601 tggtaccctc cagaccagcg ccaccgcctc cgacagtgtc ctcgctctac ctctgagggg
     3661 cttggggagg ggctggggct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3721 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3781 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3841 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3901 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3961 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4021 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4081 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4141 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4201 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     4261 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     4321 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     4381 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     4441 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     4501 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     4561 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     4621 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     4681 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     4741 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     4801 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     4861 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     4921 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     4981 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     5041 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     5101 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     5161 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     5221 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     5281 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc
     5341 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5401 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5461 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5521 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5581 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5641 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5701 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5761 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5821 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5881 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5941 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6001 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6061 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6121 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6181 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6241 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6301 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6361 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6421 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6481 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6541 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6601 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6661 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6721 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6781 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6841 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6901 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6961 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7021 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7081 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7141 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7201 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7261 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7321 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7381 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7441 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7501 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7561 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7621 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7681 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7741 ccccgaaaag tgccacctga cgtc
//
