LOCUS       V034536                 9473 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034536
VERSION     V034536
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             896..1612
                     /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             1652..4909
                     /label="Nnt(NM_001013157)"
                     /note="Nnt(NM_001013157)"
                     /gene="Nnt"
     CDS             4916..5005
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    5040..5264
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5310..5738
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5752..6081
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6148..6942
                     /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    7116..7237
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7286..7302)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7310..7326
                     /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(7334..7364)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7379..7400
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7688..8273)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8444..9304)
                     /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(9305..9409)
                     /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 ggctagcgtt taaacttaag cttggtaccg ccaccatggt
      901 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      961 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1021 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1081 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1141 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1201 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1261 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1321 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1381 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1441 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1501 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1561 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtgg
     1621 aggcagtggt ggaggcggta gcgatggatc catggcacat ctactgaaaa cggtggtgac
     1681 tggctgttta tgtccttctg ttagcaattt ggggtcctgt aaggttctac ctgggaagaa
     1741 agacttttta cgaacatttc gtacccacca ggcactgtgg tgtaaatccc cggcaaaacc
     1801 aggaattcca tataagcagt tgacagttgg ggtccccaag gagattttcc aaaatgagaa
     1861 gcgagttgcc ttgtctcctg ctggtgtcca ggccctggtc aagcagggtt ttaacgttgt
     1921 tgtggaatca ggagcaggcg aagcttccaa attcccagat gatctctaca gagcagcagg
     1981 ggcccagatc caaggaacga aggaagttct ggcttctgat ctggtggtca aagtgagggc
     2041 ccccatggtg aacccgacgt taggtgctca tgaagctgac tttttaaagc cctcaggaac
     2101 tctaattagc ttcatttacc cagcccaaaa tccagacttg ctaaataaac tctccgaaag
     2161 aaaaactaca gttttggcga tggaccaggt tccgagggtg acaatagctc agggctatga
     2221 tgctctcagc tccatggcca acatttctgg ttataaggct gttgttttag cagccaatca
     2281 ttttggaagg tttttcactg gtcagatcac agctgctgga aaagtccccc cagcgaagat
     2341 cctgatagtt ggtggtggcg ttgccgggct cgcttctgca ggggcagcaa agtcaatggg
     2401 cgcagttgtc cgaggatttg atacaagagc tgccgccttg gaacagttca aatcccttgg
     2461 tgctgaaccc ttggaggtgg acttgaagga gtctggtgag ggccaaggag gatacgcgaa
     2521 ggagatgtcc aaagagttta ttgaggcgga aatgaagctc tttgctcagc aatgcaagga
     2581 agtggacatc cttatcagca cagctctgat tccaggtaag aaagctcctg ttctatttag
     2641 taaagaaatg atcgagtcca tgaaggaagg ttcagtagtt gtggatttag ctgctgaggc
     2701 cggcggaaac tttgaaacga ctaaaccggg agaactttac gttcataagg gaatcactca
     2761 tattggctac acagacctcc ccagccgcat ggccactcag gctagcactc tgtactccaa
     2821 caacatcacc aagctcctga aggccatcag ccccgacaag gacaattttc actttgaagt
     2881 gaaggatgac tttgactttg gtacaatgag tcacgtcatt cgaggaaccg tggtgatgaa
     2941 ggatggcaat gtgattttcc cagctcccac accaaaaaat attcccaaag aagccccagc
     3001 caaacagaag accgtggctg agctggaagc agagaaggca ggcactgtct caatgtacac
     3061 gaagacactg aggacagcat ccgtgtattc agcaggtctc accgggatgc taggtttggg
     3121 catcgtggcc cccaatctag ccttttctca gatggtgacc acctttggcc tggcgggcat
     3181 tattggttac cacactgtct ggggagtgac ccctgctctc cactcaccac tgatgtctgt
     3241 gaccaatgct atctcaggtt tgactgcagt tggtgggctg gcgttgatgg gagggcattt
     3301 ttatccctcc acaacttccc agagtcttgc tgctcttgct acattcatat cgtcagtcaa
     3361 cattgcaggt ggttttctgg tgactcagag aatgttggac atgttcaagc gacccacgga
     3421 ccctccagaa tacaattacc tgtatctgct ccctggcggc accttcgtgg ggggatattt
     3481 agcagctctc tacggtggtt ataacatcga agaaatcatg tacctgggct caggtttgtg
     3541 ctgtgttggg gccctgggcg gcctttccac tcagggaaca gctcgactcg gcaatgcctt
     3601 gggcatgatt ggggttgcag gaggcctggc agctaccctt ggaggcctga agccggaccc
     3661 acagctgtta gctcagatgt ccggagcaat ggcgatgggc ggcaccatcg gactggcgat
     3721 tgcaaagcgg atccagatct ccgacctgcc tcaactggtt gctgctttcc acagtttagt
     3781 gggtctggcg gcagtgctca catgcatggc cgagtacatc gtggagtatc cacatttcgc
     3841 aatggatgcc acttcaaatt tcaccaagat tgtggcctac attggcactt acatcggcgg
     3901 agtcaccttt agtgggtctc tggttgccta cggaaaatta cagggtattc tgaagtcagc
     3961 ccctctgctg ctccctggaa ggcacgcgct caatgcaggc ttgctggctg ctagcgtggg
     4021 gggaataatc ccatttatgg ctgaccctag ttttaccaca ggcatcatgt gtctgggctc
     4081 tgtgtccgca ctctctactc tgatgggtgt gaccttgaca gctgccattg gcggtgctga
     4141 catgcctgtt gttatcactg tgctaaacag ctactccggc tgggctctgt gtgcggaggg
     4201 cttcctgctc aacaacaacc tgttgaccat tgtgggagca ctcatcggct cctcgggcgc
     4261 catcttgtcg tacatcatgt gtgtggcaat gaaccgttcc ctggcaaatg taattctcgg
     4321 aggctatggt acgacttcaa cagctggtgg aaaacccatg gaaatttccg gcacacatac
     4381 agaaatcaac ctggacaatg cagtcgagat gatccgggaa gctaacagca ttgttattac
     4441 tccaggatat gggctctgtg cagctaaagc tcagtacccc attgccgatt tggtgaagat
     4501 gcttaccgaa caaggcaaga aagtcaggtt tggaatccac ccagttgcgg gccgaatgcc
     4561 tggtcagctt aatgtgctcc tggccgaggc aggagtgccc tatgatattg tgctagaaat
     4621 ggatgagatc aacagtgatt tcccagatac cgatctggtt cttgtcattg gagctaatga
     4681 caccgtgaat tcagcggctc aggaagaccc caattctatt attgcaggca tgccagtcct
     4741 cgaggtctgg aagtccaaac aggtcattgt catgaagagg tcccttggtg ttggctacgc
     4801 tgcagtggac aatccaatct tctacaaacc taacacagcc atgcttctgg gggacgccaa
     4861 gaagacatgt gatgctctgc aggccaaagt tcgagaatcc tatcagaagc tcgaggaaca
     4921 aaaactaata tcagaggaag atttggaaca gaaactaatc agtgaagaag atttagagca
     4981 aaagttaatt tccgaggagg acttatgagg gcccgtttaa acccgctgat cagcctcgac
     5041 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5101 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5161 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5221 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     5281 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     5341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     5761 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5821 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5881 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6001 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     6061 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     6121 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     6181 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     6241 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     6301 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     6361 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     6421 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     6481 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     6541 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     6601 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6661 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6721 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6781 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6841 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6901 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6961 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     7021 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     7081 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     7141 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7201 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     7261 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7321 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7381 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7441 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7501 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7561 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7621 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8101 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8221 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8521 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8581 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8641 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8701 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8761 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8821 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8881 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8941 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9001 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9061 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9121 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9181 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9241 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9301 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9361 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9421 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
