LOCUS       V036600                 9064 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036600
VERSION     V036600
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             874..1590
                     /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"
     CDS             1642..4554
                     /label="Osmr(NM_011019)"
                     /note="Osmr(NM_011019)"
                     /gene="Osmr"
     polyA_signal    4631..4855
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4901..5329
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5343..5672
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5739..6533
                     /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    6707..6828
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6877..6893)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6901..6917
                     /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(6925..6955)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6970..6991
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7279..7864)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8035..8895)
                     /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(8896..9000)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggctttc tctgtggtcc ttcatccagc cttcctcctg
     1681 gcagtgctgt ccctgagggc atcccgaagc gaagtcttgg aggagccttt accattgact
     1741 cctgagatac ataaagtttc ttttcaattg aaacttcaag aagtgaattt agaatggact
     1801 gtcccagccc ttactcatga agaattaaac atgatatttc agatagagat cagtagactg
     1861 aacatatcca acaccatctg ggtggagaat tatagcacca ctgtgaagcg tgaagaagct
     1921 gtgcgttgga actggacgtc tgatatccct ttggagtgtg tcaaacattt cataagaatc
     1981 agggctctgg tagatgacac caagtccctt ccacagagtt cctggggcaa ctggagttcc
     2041 tggaaagaag ttaatgcaaa ggtttccgtt gaacctgata aatcattaat atttcctaaa
     2101 gacaaagtgt tggaagaagg ctccaatgtc accatctgtc tgatgtatgg gcagaatgta
     2161 tataatgtat cctgtaagtt gcaagatgag ccaatccatg gagaacaact tgattcccac
     2221 gtgtcattat taaaattgaa caatgtagtt ttccttagtg acacagggac aaacatcaat
     2281 tgtcaagcca cgaagggtcc taaaagaata tttggtactg ttctctttgt ctcgaaagtg
     2341 ctcgaggaac ctaagaatgt ttcctgtgaa acccgagact ttaagacttt ggactgttca
     2401 tgggaacctg gggtagatac gactttgact tggcgtaaac aaagattcca aaactacact
     2461 ttatgtgaat cgttctctaa gagatgtgag gtttctaact acaggaactc ctatacctgg
     2521 caaatcactg aaggctcaca ggaaatgtat aactttactc tcacagctga aaaccaacta
     2581 aggaaaagaa gtgtcaacat taattttaac ctgacccata gagttcatcc aaaggctccg
     2641 caggacgtca cccttaaaat tataggtgct acaaaagcca acatgacttg gaaggttcac
     2701 tcccatggaa acaactacac acttttgtgt caggttaaac tccaatatgg agaagtgatt
     2761 catgagcaca atgtttctgt ccacatgagc gcaaactacc tcttcagtga tctggatcca
     2821 gacacaaagt acaaggcttt tgtgcgctgt gcaagtgcca accacttctg gaaatggagc
     2881 gactggaccc aaaaagagtt cagcacaccc gagactgctc cctcacaggc tcttgatgta
     2941 tggagacaag tgtggtcgga gaatggaaga cgcattgtga ctttattctg gaagccacta
     3001 ttaaaatcac aggccaatgg caaaatcata tcctataata tagttgtaga aaatgaagcc
     3061 aaaccaactg agtcagaaca ctactgtgtc tgggcaccag ccctcagcac aaacctgagc
     3121 cttgacctgc aaccttacaa gattcgcatc acagccaaca acagcatggg ggcatctcct
     3181 gagtccttga tggtcctttc taatgattct ggacacgaag aggtcaagga aaagacaatt
     3241 aaaggtataa aggatgcatt caatatttct tgggagcccg tatctggaga cacgatgggc
     3301 tatgttgtgg actggtgtgc acattcccag gaccaacgct gtgatttgca gtggaagaac
     3361 cttggtccca ataccacaag caccaccatc acctcagatg attttaaacc aggcgtccgt
     3421 tacaacttca gaatttttga aaggtctgtg gaacacaaag ctcggttagt agagaaacaa
     3481 agaggataca cccaggaact ggctcctttg gtgaatccaa aagtggagat tccttactcg
     3541 acccctaact ccttcgttct aagatggcca gattatgaca gcgacttcca ggctggtttt
     3601 ataaaagggt acctcgtgta tgtgaaatcc aaggagatgc agtgcaacca accctgggaa
     3661 aggaccctcc ttccagataa ttcagtcctc tgtaaatacg acatcaatgg ctcagagaca
     3721 aagacactca ccgtggaaaa ccttcagcca gagtccctct atgagttttt cgtcactccg
     3781 tacaccagcg ctggcccagg acccaatgaa acgttcacaa aggtcacaac tccagatgca
     3841 cgctcccaca tgctgctgca gatcatacta cccatgaccc tctgcgtctt gctcagcatc
     3901 attgtctgct actggaaaag tcagtgggtg aaggagaagt gctaccctga cattcccaat
     3961 ccgtacaaga gcagcattct gtcactcata aaatccaaga agaatcctca cttaataatg
     4021 aatgtcaaag actgcattcc agatgtcctt gaagtgataa acaaagcaga aggcagcaag
     4081 acacagtgtg taggctctgg gaaacttcac attgaagatg tacccactaa gccgccaatc
     4141 gtgccaacag aaaaggattc ctcagggcct gtgccctgca tcttctttga gaattttact
     4201 tacgatcagt cagcttttga ctctggttcc catggcctca ttccaggtcc cctaaaagac
     4261 acagcacacc aacttggact attggctcca cctaacaagt tccagaacgt attaaaaaat
     4321 gactacatga agcccctggt cgaaagtcca actgaagaaa ctagcttgat ttatgtgtca
     4381 cagctggctt cacccatgtg cggagacaag gacacgcttg ccacagaacc acccgtgcca
     4441 gtgcatggtt cagagtataa aaggcaaatg gtagttcccg ggagcctcgc atcaccttct
     4501 ctgaaggagg ataacagctt gacctcaacg gtcctcttag gccaaggtga acagtaagga
     4561 tcctgatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     4621 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4681 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4741 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4801 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4861 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4921 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4981 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5041 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5101 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5161 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5221 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5281 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5341 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5401 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5461 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5521 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5581 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5641 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5701 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5761 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5821 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5881 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5941 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6001 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6061 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6121 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6181 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6241 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6301 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6361 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6421 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6481 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6541 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6601 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6661 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6721 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6781 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6841 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6901 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6961 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7021 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7081 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7681 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7801 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8101 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8161 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8221 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8281 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8341 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8401 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8461 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8521 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8581 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8641 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8701 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8761 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8821 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8881 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8941 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9001 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     9061 tggt
//
