LOCUS       V027928                 8469 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027928
VERSION     V027928
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"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..3272
                     /label="ANO9(NM_001012302)"
                     /note="ANO9(NM_001012302)"
                     /gene="ANO9"
     CDS             3312..4031
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4069..4293
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4339..4767
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4781..5110
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5177..5971
                     /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    6145..6266
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6315..6331)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6339..6355
                     /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(6363..6393)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6408..6429
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6717..7302)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7473..8333)
                     /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(8334..8438)
                     /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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatgc agggcgaaga gagcctccgg atcctggtgg
      961 agcccgaagg ggacagcttc ccgctgatgg agatcagcac ctgtgagacc gaggcctccg
     1021 agcagtggga ctatgtcctc gtggcccaac gtcacaccca gagagacccc cggcaggcgc
     1081 ggcagcaaca gttcctggag gagctcagga gaaagggctt ccacattaag gtgatccggg
     1141 accagaaaca ggtcttcttt gggatccgtg ctgacaacag tgtctttggc ctgtaccgca
     1201 ctctcctcct ggagcctgag gggcctgccc cccacgccga gctggccgcg ccgaccacca
     1261 tcccggtcac cacgagtctc agaatccgaa tcgtgaactt cgttgtcatg aacaacaaga
     1321 cctcggctgg tgagaccttc gaggatctga tgaaggacgg ggtctttgag gccaggttcc
     1381 ccctgcacaa gggggaggga cgcctgaaga agacgtgggc gcggtggaga cacatgttcc
     1441 gggagcagcc agttgatgaa atcaggaact actttgggga aaaggtggcc ctgtacttcg
     1501 tctggctggg ctggtacacc tacatgctgg tgccggccgc cctgacgggc ctcttagtct
     1561 ttctgagcgg attctcgctg tttgaggcca gccagatcag caaggagatc tgtgaggccc
     1621 acgacatcct catgtgtccc ctcggcgacc acagccgcag gtaccagcgg ctctcggaaa
     1681 cctgcacttt tgccaagctc acccacctct ttgacaatga tggcacggtg gtgttcgcca
     1741 tcttcatggc tctctgggcc acggtgttcc tggagatctg gaagcggcag cgcgcccgcg
     1801 tggtcctgca ctgggacctg tacgtgtggg acgaggaaca ggaggaaatg gcacttcagc
     1861 tcattaactg ccccgactac aagctccggc cataccagca ctcctaccta cgcagcaccg
     1921 tcatcctcgt cctgaccctg ctcatgatct gcctcatgat cggcatggcc cacgtcctgg
     1981 tggtctaccg cgtcctggcc tccgcgctct tcagcagctc ggccgtgccc ttcctggagg
     2041 agcaggtgac cacggccgtg gtggtgaccg gggctctggt gcactatgtg accatcatca
     2101 tcatgaccaa gatcaacagg tgcgtggccc tgaagctttg tgacttcgag atgcccagga
     2161 ccttctcgga gcgagagagc aggttcacca tccgcttctt cacactgcag ttcttcaccc
     2221 atttctcgtc tctcatctac atcgccttca tcctgggcag gatcaacggc caccccggga
     2281 agtccacgcg cctggcgggc ttgtggaagc tggaagagtg ccacgccagc ggctgcatga
     2341 tggacctctt cgtgcagatg gccatcatca tgggcctgaa gcagacgctc agcaactgcg
     2401 tcgagtacct ggtcccgtgg gtgacccaca agtgccgctc tctgcgggcc tccgagtccg
     2461 ggcacctgcc ccgggacccc gagctcaggg actggcggcg caactacctt ctgaacccgg
     2521 tcaacacctt cagcctgttc gacgagttca tggagatgat gatccagtac ggcttcacca
     2581 ccatcttcgt ggccgccttc ccgctggcgc cgctgctcgc gctcttcagc aacctcgtgg
     2641 agatccgcct ggacgccatc aagatggtct ggttgcagcg gcgcctggtg ccgcgcaagg
     2701 ccaaggacat cgggacctgg ctgcaggtgc tggagaccat cggtgtgctg gcggtcattg
     2761 ccaatgggat ggtcattgcc ttcacatctg agttcatccc ccgagtggtc tacaagtacc
     2821 gctatagccc atgcctgaaa gaaggcaact ctactgtcga ctgcctcaag ggctacgtca
     2881 accacagcct gtccgtcttc cacaccaagg acttccagga ccctgatggg attgagggct
     2941 cagaaaacgt gactctgtgc agatacaggg actaccgcaa tccccccgat tacaacttct
     3001 ccgagcagtt ctggttcctc ctggccatcc gcctggcctt cgtcatcctc tttgagcacg
     3061 tggccttgtg catcaagctc atcgccgcct ggttcgtgcc cgacatccct cagtcggtga
     3121 agaacaaggt tctggaggtg aagtaccaga ggctgcgtga gaagatgtgg catggaaggc
     3181 agaggctggg tggggtgggg gcaggctctc ggcccccaat gcctgcccat cccaccccag
     3241 catccatctt cagtgccagg agcacagacg tgggctcgag tggcggtgga ggaagcggcg
     3301 gaggtggatc tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3361 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     3421 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3481 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3541 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3601 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3661 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3721 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3781 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3841 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3901 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3961 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     4021 tgtacaagta gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     4081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4321 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4921 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5041 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5101 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5161 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5221 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5281 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5341 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5401 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5461 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5521 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5581 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5641 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5701 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5761 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5821 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5881 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5941 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6001 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6061 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6121 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6181 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6241 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8461 cctgacgtc
//
