LOCUS       V020975                 9115 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020975
VERSION     V020975
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             649..1365
                     /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             1387..3792
                     /label="FGFR4(NM_002011)"
                     /note="FGFR4(NM_002011)"
                     /gene="FGFR4"
     CDS             3793..3858
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3891..4479
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4545..4593
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4795..5223
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5237..5566
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5633..6427
                     /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    6603..6724
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6773..6789)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6797..6813
                     /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(6821..6851)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6866..6887
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7175..7763)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7934..8794)
                     /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(8795..8899)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccatgc ggctgctgct ggccctgttg ggggtcctgc tgagtgtgcc tgggcctcca
     1441 gtcttgtccc tggaggcctc tgaggaagtg gagcttgagc cctgcctggc tcccagcctg
     1501 gagcagcaag agcaggagct gacagtagcc cttgggcagc ctgtgcgtct gtgctgtggg
     1561 cgggctgagc gtggtggcca ctggtacaag gagggcagtc gcctggcacc tgctggccgt
     1621 gtacggggct ggaggggccg cctagagatt gccagcttcc tacctgagga tgctggccgc
     1681 tacctctgcc tggcacgagg ctccatgatc gtcctgcaga atctcacctt gattacaggt
     1741 gactccttga cctccagcaa cgatgatgag gaccccaagt cccataggga cccctcgaat
     1801 aggcacagtt acccccagca agcaccctac tggacacacc cccagcgcat ggagaagaaa
     1861 ctgcatgcag tacctgcggg gaacaccgtc aagttccgct gtccagctgc aggcaacccc
     1921 acgcccacca tccgctggct taaggatgga caggcctttc atggggagaa ccgcattgga
     1981 ggcattcggc tgcgccatca gcactggagt ctcgtgatgg agagcgtggt gccctcggac
     2041 cgcggcacat acacctgcct ggtagagaac gctgtgggca gcatccgcta taactacctg
     2101 ctagatgtgc tggagcggtc cccgcaccgg cccatcctgc aggccgggct cccggccaac
     2161 accacagccg tggtgggcag cgacgtggag ctgctgtgca aggtgtacag cgatgcccag
     2221 ccccacatcc agtggctgaa gcacatcgtc atcaacggca gcagcttcgg agccgacggt
     2281 ttcccctatg tgcaagtcct aaagactgca gacatcaata gctcagaggt ggaggtcctg
     2341 tacctgcgga acgtgtcagc cgaggacgca ggcgagtaca cctgcctcgc aggcaattcc
     2401 atcggcctct cctaccagtc tgcctggctc acggtgctgc cagaggagga ccccacatgg
     2461 accgcagcag cgcccgaggc caggtatacg gacatcatcc tgtacgcgtc gggctccctg
     2521 gccttggctg tgctcctgct gctggccggg ctgtatcgag ggcaggcgct ccacggccgg
     2581 cacccccgcc cgcccgccac tgtgcagaag ctctcccgct tccctctggc ccgacagttc
     2641 tccctggagt caggctcttc cggcaagtca agctcatccc tggtacgagg cgtgcgtctc
     2701 tcctccagcg gccccgcctt gctcgccggc ctcgtgagtc tagatctacc tctcgaccca
     2761 ctatgggagt tcccccggga caggctggtg cttgggaagc ccctaggcga gggctgcttt
     2821 ggccaggtag tacgtgcaga ggcctttggc atggaccctg cccggcctga ccaagccagc
     2881 actgtggccg tcaagatgct caaagacaac gcctctgaca aggacctggc cgacctggtc
     2941 tcggagatgg aggtgatgaa gctgatcggc cgacacaaga acatcatcaa cctgcttggt
     3001 gtctgcaccc aggaagggcc cctgtacgtg atcgtggagt ccgccgccaa gggaaacctg
     3061 cgggagttcc tgcgggcccg gcgcccccca ggccccgacc tcagccccga cggtcctcgg
     3121 agcagtgagg ggccgctctc cttcccagtc ctggtctcct gcgcctacca ggtggcccga
     3181 ggcatgcagt atctggagtc ccggaagtgt atccaccggg acctggctgc ccgcaatgtg
     3241 ctggtgactg aggacaatgt gatgaagatt gctgactttg ggctggcccg cggcgtccac
     3301 cacattgact actataagaa aaccagcaac ggccgcctgc ctgtgaagtg gatggcgccc
     3361 gaggccttgt ttgaccgggt gtacacacac cagagtgacg tgtggtcttt tgggatcctg
     3421 ctatgggaga tcttcaccct cgggggctcc ccgtatcctg gcatcccggt ggaggagctg
     3481 ttctcgctgc tgcgggaggg acatcggatg gaccgacccc cacactgccc cccagagctg
     3541 tacgggctga tgcgtgagtg ctggcacgca gcgccctccc agaggcctac cttcaagcag
     3601 ctggtggagg cgctggacaa ggtcctgctg gccgtctctg aggagtacct cgacctccgc
     3661 ctgaccttcg gaccctattc cccctctggt ggggacgcca gcagcacctg ctcctccagc
     3721 gattctgtct tcagccacga ccccctgcca ttgggatcca gctccttccc cttcgggtct
     3781 ggggtgcaga cagactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3841 aaggatgacg atgacaagta gctcgagtct agagggcccg atatctcgac aatcaacctc
     3901 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3961 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4021 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4081 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4141 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4201 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4261 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4321 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4381 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4441 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac
     4501 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4561 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4621 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4681 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4741 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     4801 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4861 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4921 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4981 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5041 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5101 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5161 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5221 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5281 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5401 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5461 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5521 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5581 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5641 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5701 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5761 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5821 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5881 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5941 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6001 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6061 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6121 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6181 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6241 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6301 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6361 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6421 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     6481 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6541 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6601 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6721 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7981 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8041 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8101 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8161 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8221 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8281 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8341 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8401 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8461 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8521 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8581 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8641 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8701 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8761 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8821 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8881 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8941 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     9001 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     9061 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
