LOCUS       V027290                 7601 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027290
VERSION     V027290
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"
     CDS             1008..3464
                     /label="ADAM9(NM_003816)"
                     /note="ADAM9(NM_003816)"
                     /gene="ADAM9"
     CDS             3465..3512
                     /codon_start=1
                     /product="two tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDK"
                     /label="2xFLAG"
     polyA_signal    3559..3783
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4067..4114
                     /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             4122..5144
                     /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    5274..5395
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5444..5460)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5468..5484
                     /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(5492..5522)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5537..5558
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5846..6434)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6605..7465)
                     /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(7466..7570)
                     /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 gggtctggcg
     1021 cgcgctttcc ctcggggacc cttcgtgtcc ggtggttgct gttgcttggc ctggtgggcc
     1081 cagtcctcgg tgcggcgcgg ccaggctttc aacagacctc acatctttct tcttatgaaa
     1141 ttataactcc ttggagatta actagagaaa gaagagaagc ccctaggccc tattcaaaac
     1201 aagtatctta tgttattcag gctgaaggaa aagagcatat tattcacttg gaaaggaaca
     1261 aagacctttt gcctgaagat tttgtggttt atacttacaa caaggaaggg actttaatca
     1321 ctgaccatcc caatatacag aatcattgtc attatcgggg ctatgtggag ggagttcata
     1381 attcatccat tgctcttagc gactgttttg gactcagagg attgctgcat ttagagaatg
     1441 cgagttatgg gattgaaccc ctgcagaaca gctctcattt tgagcacatc atttatcgaa
     1501 tggatgatgt ctacaaagag cctctgaaat gtggagtttc caacaaggat atagagaaag
     1561 aaactgcaaa ggatgaagag gaagagcctc ccagcatgac tcagctactt cgaagaagaa
     1621 gagctgtctt gccacagacc cggtatgtgg agctgttcat tgtcgtagac aaggaaaggt
     1681 atgacatgat gggaagaaat cagactgctg tgagagaaga gatgattctc ctggcaaact
     1741 acttggatag tatgtatatt atgttaaata ttcgaattgt gctagttgga ctggagattt
     1801 ggaccaatgg aaacctgatc aacatagttg ggggtgctgg tgatgtgctg gggaacttcg
     1861 tgcagtggcg ggaaaagttt cttatcacac gtcggagaca tgacagtgca cagctagttc
     1921 taaagaaagg ttttggtgga actgcaggaa tggcatttgt gggaacagtg tgttcaagga
     1981 gccacgcagg cgggattaat gtgtttggac aaatcactgt ggagacattt gcttccattg
     2041 ttgctcatga attgggtcat aatcttggaa tgaatcacga tgatgggaga gattgttcct
     2101 gtggagcaaa gagctgcatc atgaattcag gagcatcggg ttccagaaac tttagcagtt
     2161 gcagtgcaga ggactttgag aagttaactt taaataaagg aggaaactgc cttcttaata
     2221 ttccaaagcc tgatgaagcc tatagtgctc cctcctgtgg taataagttg gtggacgctg
     2281 gggaagagtg tgactgtggt actccaaagg aatgtgaatt ggacccttgc tgcgaaggaa
     2341 gtacctgtaa gcttaaatca tttgctgagt gtgcatatgg tgactgttgt aaagactgtc
     2401 ggttccttcc aggaggtact ttatgccgag gaaaaaccag tgagtgtgat gttccagagt
     2461 actgcaatgg ttcttctcag ttctgtcagc cagatgtttt tattcagaat ggatatcctt
     2521 gccagaataa caaagcctat tgctacaacg gcatgtgcca gtattatgat gctcaatgtc
     2581 aagtcatctt tggctcaaaa gccaaggctg cccccaaaga ttgtttcatt gaagtgaatt
     2641 ctaaaggtga cagatttggc aattgtggtt tctctggcaa tgaatacaag aagtgtgcca
     2701 ctgggaatgc tttgtgtgga aagcttcagt gtgagaatgt acaagagata cctgtatttg
     2761 gaattgtgcc tgctattatt caaacgccta gtcgaggcac caaatgttgg ggtgtggatt
     2821 tccagctagg atcagatgtt ccagatcctg ggatggttaa cgaaggcaca aaatgtggtg
     2881 ctggaaagat ctgtagaaac ttccagtgtg tagatgcttc tgttctgaat tatgactgtg
     2941 atgttcagaa aaagtgtcat ggacatgggg tatgtaatag caataagaat tgtcactgtg
     3001 aaaatggctg ggctccccca aattgtgaga ctaaaggata cggaggaagt gtggacagtg
     3061 gacctacata caatgaaatg aatactgcat tgagggacgg acttctggtc ttcttcttcc
     3121 taattgttcc ccttattgtc tgtgctattt ttatcttcat caagagggat caactgtgga
     3181 gaagctactt cagaaagaag agatcacaaa catatgagtc agatggcaaa aatcaagcaa
     3241 acccttctag acagccgggg agtgttcctc gacatgtttc tccagtgaca cctcccagag
     3301 aagttcctat atatgcaaac agatttgcag taccaaccta tgcagccaag caacctcagc
     3361 agttcccatc aaggccacct ccaccacaac cgaaagtatc atctcaggga aacttaattc
     3421 ctgcccgtcc tgctcctgca cctcctttat atagttccct cactgactac aaggacgacg
     3481 atgacaagga ttacaaggat gacgacgata agtgactcga gtctagaggg cccgtttaaa
     3541 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3601 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3661 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3721 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3781 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3841 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3901 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3961 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4021 acctcgaccc caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc
     4081 gaagttccta ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg
     4141 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     4201 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     4261 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     4321 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     4381 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     4441 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     4501 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     4561 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     4621 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     4681 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     4741 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     4801 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     4861 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     4921 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     4981 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     5041 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     5101 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
     5161 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5221 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5281 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5341 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5401 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5461 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5521 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5581 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5641 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6601 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
