LOCUS       V047659                 8451 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047659
VERSION     V047659
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..3191
                     /label="Myb(NM_001198914)"
                     /note="Myb(NM_001198914)"
                     /gene="Myb"
     CDS             3198..3263
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             3294..4013
                     /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    4051..4275
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4321..4749
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4763..5092
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5159..5953
                     /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    6127..6248
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6297..6313)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6321..6337
                     /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(6345..6375)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6390..6411
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6699..7284)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7455..8315)
                     /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(8316..8420)
                     /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 gccaccatgg cccggagacc ccgacacagc atctacagta
      961 gcgatgaaga tgatgaagac attgagatgt gtgaccatga ctacgatggg ctgctgccca
     1021 aatctggaaa gcgtcacttg gggaaaacta ggtggacaag ggaagaggat gagaagctga
     1081 agaagctggt ggaacagaac ggaacagacg actggaaagt cattgccaat tatctgccca
     1141 accggacaga tgtgcagtgc caacaccggt ggcagaaagt gctgaaccct gaactcatca
     1201 aaggtccctg gaccaaagaa gaagatcaga gagtcataga gcttgtccag aaatatggtc
     1261 cgaagcgttg gtctgttatt gccaagcact taaaagggag aattggaaag cagtgtcggg
     1321 agaggtggca caaccatttg aatccagaag ttaagaaaac ctcctggaca gaagaggagg
     1381 acagaatcat ttaccaggca cacaagcgtc tggggaacag atgggcagag atcgcaaagc
     1441 tgctgcccgg acggactgat aatgctatca agaaccactg gaattccacc atgcgtcgca
     1501 aggtggaaca ggaaggctac ctgcaggagc cttccaaagc cagccagacg ccagtggcca
     1561 cgagcttcca gaagaacaat catttgatgg ggtttgggca tgcctcacct ccatctcagc
     1621 tctctccaag tggccagtcc tccgtcaaca gcgaatatcc ctattaccac atcgccgaag
     1681 cacaaaacat ctccagtcac gttccctatc ctgtcgcatt gcatgttaat atagtcaacg
     1741 tccctcagcc ggctgcggca gccatccaga gacactataa cgacgaagac cctgagaagg
     1801 aaaagcgaat aaaggagctg gagttgctcc tgatgtcaac agagaacgag ctgaagggac
     1861 agcaggcatt accaacacag aaccacactt gcagctaccc cgggtggcac agcacctcca
     1921 ttgtggacca gaccagacct catggggata gtgcacctgt ttcctgtttg ggagaacacc
     1981 atgccacccc atctctgcct gcagatcccg gctccctacc tgaagaaagt gcctcaccag
     2041 caaggtgcat gatcgtccac cagggcacca ttctggacaa tgttaagaac ctcttagaat
     2101 ttgcagaaac actccagttt atagattctg attcttcgtg gtgtgatctc agcagttttg
     2161 aattctctga agaagcggca gctttttcac ctagccagca gcccacaggc aaagccttcc
     2221 agcttcagca aagagagggc catgggacta gatctgcagg agagcctagc ctgagggtga
     2281 ccaggcgagt gctgagcgag gcatccctcg gcccagactc accccaagcg aggcacagca
     2341 aggttccgct ggtcgtccta cgaaaaaggc ggggccaggc cagcccccta gccgctggag
     2401 agcctagccc ctccctcttt gctgacgtca tcagctcaac tctcaagcgt tcccctgtca
     2461 aaagcctacc cttctctccc tcgcagttct tgaacacttc cagcaaccat gaaagctcgg
     2521 gcttagatgc acctacctta ccctccactc ctctcattgg tcacaaactg acaccatgtc
     2581 gagaccagac tgtgaaaacc cagaaggaaa attccatctt tagaactcca gctatcaaaa
     2641 ggtcaatcct cgaaagctct cctcgaactc ccacaccatt caaacatgcc cttgcagctc
     2701 aagaaattaa atacggtccc ctgaagatgc tacctcagac cccctcccat gcagtggagg
     2761 acctacaaga tgtgattaag caggaatcgg atgaatctgg aattgtggct gagtttcaag
     2821 agagtggacc accgttactg aaaaaaatca agcaggaggt ggagtcgcca actgagaaat
     2881 cgggaaactt cttctgctca aaccactggg cagagaacag cctgagcacc cagctgttct
     2941 cgcaggcgtc tcctgtggca gatgccccaa atattcttac aagctctgtt ttaatgacac
     3001 ctgtatcaga agatgaagac aatgtcctca aagcctttac cgtacctaag aacaggcccc
     3061 tggtgggtcc cttgcagcca tgcagtggtg cctgggagcc agcatcctgt gggaagacag
     3121 aggaccagat gacggcctcc ggtccggctc ggaaatacgt gaacgcgttc tcagctcgaa
     3181 ctctggtcat gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg
     3241 actacaagga tgacgatgac aagggcggtg gaggaagcgg cggaggtgga tctatggtga
     3301 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3361 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3421 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3481 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3541 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3601 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3661 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3721 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     3781 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     3841 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3901 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3961 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tagtctagag
     4021 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4081 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4141 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4201 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4261 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     4321 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4381 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4441 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4501 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4561 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4621 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4681 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4741 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4801 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4921 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4981 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     5041 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     5101 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     5161 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5221 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5281 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5341 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5401 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5461 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5521 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5581 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5641 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5701 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5761 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5821 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5881 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5941 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6001 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6061 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6121 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6241 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     6301 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6361 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6421 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6481 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6541 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6601 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6661 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6721 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6781 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6841 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6901 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6961 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7021 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7081 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7141 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7201 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     7261 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7321 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7381 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7441 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7501 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7561 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7621 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7681 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7741 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7801 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7861 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7921 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7981 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8041 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8101 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8161 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8221 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8281 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8341 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8401 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
