LOCUS       V023204                 8410 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023204
VERSION     V023204
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             872..3115
                     /label="ADAM10(NM_001110)"
                     /note="ADAM10(NM_001110)"
                     /gene="ADAM10"
     CDS             3167..3877
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     polyA_signal    3977..4201
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4247..4675
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4689..5018
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5085..5879
                     /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    6053..6174
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6223..6239)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6247..6263
                     /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(6271..6301)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6316..6337
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6625..7210)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7381..8241)
                     /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(8242..8346)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccac catggtgttg ctgagagtgt taattctgct
      901 cctctcctgg gcggcgggga tgggaggtca gtatgggaat cctttaaata aatatatcag
      961 acattatgaa ggattatctt acaatgtgga ttcattacac caaaaacacc agcgtgccaa
     1021 aagagcagtc tcacatgaag accaattttt acgtctagat ttccatgccc atggaagaca
     1081 tttcaaccta cgaatgaaga gggacacttc ccttttcagt gatgaattta aagtagaaac
     1141 atcaaataaa gtacttgatt atgatacctc tcatatttac actggacata tttatggtga
     1201 agaaggaagt tttagccatg ggtctgttat tgatggaaga tttgaaggat tcatccagac
     1261 tcgtggtggc acattttatg ttgagccagc agagagatat attaaagacc gaactctgcc
     1321 atttcactct gtcatttatc atgaagatga tattaactat ccccataaat acggtcctca
     1381 ggggggctgt gcagatcatt cagtatttga aagaatgagg aaataccaga tgactggtgt
     1441 agaggaagta acacagatac ctcaagaaga acatgctgct aatggtccag aacttctgag
     1501 gaaaaaacgt acaacttcag ctgaaaaaaa tacttgtcag ctttatattc agactgatca
     1561 tttgttcttt aaatattacg gaacacgaga agctgtgatt gcccagatat ccagtcatgt
     1621 taaagcgatt gatacaattt accagaccac agacttctcc ggaatccgta acatcagttt
     1681 catggtgaaa cgcataagaa tcaatacaac tgctgatgag aaggacccta caaatccttt
     1741 ccgtttccca aatattggtg tggagaagtt tctggaattg aattctgagc agaatcatga
     1801 tgactactgt ttggcctatg tcttcacaga ccgagatttt gatgatggcg tacttggtct
     1861 ggcttgggtt ggagcacctt caggaagctc tggaggaata tgtgaaaaaa gtaaactcta
     1921 ttcagatggt aagaagaagt ccttaaacac tggaattatt actgttcaga actatgggtc
     1981 tcatgtacct cccaaagtct ctcacattac ttttgctcac gaagttggac ataactttgg
     2041 atccccacat gattctggaa cagagtgcac accaggagaa tctaagaatt tgggtcaaaa
     2101 agaaaatggc aattacatca tgtatgcaag agcaacatct ggggacaaac ttaacaacaa
     2161 taaattctca ctctgtagta ttagaaatat aagccaagtt cttgagaaga agagaaacaa
     2221 ctgttttgtt gaatctggcc aacctatttg tggaaatgga atggtagaac aaggtgaaga
     2281 atgtgattgt ggctatagtg accagtgtaa agatgaatgc tgcttcgatg caaatcaacc
     2341 agagggaaga aaatgcaaac tgaaacctgg gaaacagtgc agtccaagtc aaggtccttg
     2401 ttgtacagca cagtgtgcat tcaagtcaaa gtctgagaag tgtcgggatg attcagactg
     2461 tgcaagggaa ggaatatgta atggcttcac agctctctgc ccagcatctg accctaaacc
     2521 aaacttcaca gactgtaata ggcatacaca agtgtgcatt aatgggcaat gtgcaggttc
     2581 tatctgtgag aaatatggct tagaggagtg tacgtgtgcc agttctgatg gcaaagatga
     2641 taaagaatta tgccatgtat gctgtatgaa gaaaatggac ccatcaactt gtgccagtac
     2701 agggtctgtg cagtggagta ggcacttcag tggtcgaacc atcaccctgc aacctggatc
     2761 cccttgcaac gattttagag gttactgtga tgttttcatg cggtgcagat tagtagatgc
     2821 tgatggtcct ctagctaggc ttaaaaaagc aatttttagt ccagagctct atgaaaacat
     2881 tgctgaatgg attgtggctc attggtgggc agtattactt atgggaattg ctctgatcat
     2941 gctaatggct ggatttatta agatatgcag tgttcatact ccaagtagta atccaaagtt
     3001 gcctcctcct aaaccacttc caggcacttt aaagaggagg agacctccac agcccattca
     3061 gcaaccccag cgtcagcggc cccgagagag ttatcaaatg ggacacatga gacgcggcgg
     3121 cggagggagt ggaggagggg gaagcggagg cggagggagt ggtaccatgg tgagcaaggg
     3181 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     3241 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3301 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3361 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3421 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3481 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3541 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     3601 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3661 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3721 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     3781 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     3841 ccactccacc ggcggcatgg acgagctgta caagtaggga tccactagtc cagtgtggtg
     3901 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3961 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4021 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4081 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4141 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4201 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4261 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4321 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4381 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4441 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4501 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4561 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4621 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4681 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4861 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4921 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4981 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5041 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5101 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5161 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5221 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5281 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5341 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5401 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5461 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5521 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5581 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5641 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5701 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5761 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5821 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5881 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5941 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6001 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6061 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6121 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6181 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6361 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6421 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7141 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7201 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7261 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7321 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7381 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7441 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7501 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7561 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7621 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7681 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7741 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7801 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7861 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7921 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7981 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8041 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8101 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8161 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8221 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8281 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8341 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8401 cccctatggt
//
