LOCUS       V029036                 8758 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029036
VERSION     V029036
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..3468
                     /label="FBXL19(NM_001099784)"
                     /note="FBXL19(NM_001099784)"
                     /gene="FBXL19"
     misc_feature    3534..4122
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4188..4236
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4438..4866
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4880..5209
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5276..6070
                     /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    6246..6367
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6416..6432)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6440..6456
                     /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(6464..6494)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6509..6530
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6818..7406)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7577..8437)
                     /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(8438..8542)
                     /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 ggtaccatgg gtatgaaagt ccccggaaag ggggaatctg ggccctcggc gttgctgacg
     1441 cccccaatgt cgtcgagcag ccgggggccg ggggccggag cgcgccgacg ccgaacccgc
     1501 tgccgccgct gccgggcctg tgtgcgaact gagtgcgggg attgccactt ctgccgagac
     1561 atgaagaagt tcggggggcc cgggcgcatg aagcagtcgt gcctgctccg gcagtgcact
     1621 gcccccgtgc tcccacacac agctgtgtgc ctcttgtgtg gggaggctgg gaaggaggac
     1681 acggtggagg gagaggaaga gaaatttggt ttgagcctca tggagtgtac aatctgcaac
     1741 gagatcgtcc accccggctg cctgaagatg gggaaggctg agggtgtcat caatgcagag
     1801 atccccaact gctgggagtg ccctcgctgc acccaggaag gccgcaccag caaggattca
     1861 ggtgaggggc ctggccgccg tagggccgac aacggcgagg agggcgccag cttggggagc
     1921 ggatggaagc tgacagagga gccaccgctt ccaccgcccc cgcccaggcg caagggcccc
     1981 ctgcctgccg ggcccccccc ggaggacgtg cctgggcccc ccaaacgaaa ggaaagggag
     2041 gcagggaatg agcctcccac cccaaggaaa aaggtgaaag gaggccgaga gaggcacctg
     2101 aagaaggtgg gtggagacgc ctgcctcctc cgaggatcgg acccaggcgg cccgggcctg
     2161 ctgcccccca gggttctgaa tccgagccag gctttctcat cctgccaccc tgggctccct
     2221 cccgagaact gggagaaacc aaagccgcct ttggcctctg cagagggccc agcggtgccg
     2281 tccccgtccc cgcagaggga gaagctagag cgtttcaagc ggatgtgcca gctgctggaa
     2341 cgggtgcctg acacctcctc ttcctcctcg gactcagact ccgactccga ctcttcgggc
     2401 acatcgctga gtgaggacga agcccccggc gaggcccgga atgggcgacg gccagcccgg
     2461 ggcagctctg gcgagaagga gaaccgtggg gggcggcggg ctgtgcgccc tggcagtggg
     2521 gggcccctac tcagctggcc cctgggccca gccccaccac cccggcctcc acagctggag
     2581 cggcacgtgg tgcggccccc gcctcgaagc cctgagcccg acacactccc cttggctgct
     2641 ggatccgacc accccctgcc ccgggccgcc tggcttcgcg tcttccagca cctcgggccg
     2701 cgggagctgt gtatctgcat gcgagtctgc cgaacttgga gccgctggtg ctatgacaag
     2761 cgtctgtggc ctcgaatgga cctgagccgg cggaagtcac tgaccccgcc catgctcagt
     2821 ggtgtggttc gccgccagcc ccgtgccctg gacctcagct ggacaggtgt ctccaagaag
     2881 cagctcatgt ggcttctgaa ccgactacaa ggcctgcagg agctggtgct ctctggctgc
     2941 tcctggctct ctgtctctgc cctgggctca gccccactgc cagccctgcg gctcctggac
     3001 ctccgctgga tcgaggatgt taaagactcc cagctccggg agttgctgct gcctccacca
     3061 gacaccaaac cagggcaaac agagagccgt ggtcggctgc agggggtggc agaactgcgt
     3121 ctggcaggtt tggagctgac agatgcctcc ctgcgtctcc tgctgcgtca cgcaccccag
     3181 ctgagcgccc tggacctgag ccactgcgcc cacgtcgggg accccagtgt tcacctcctc
     3241 acggccccca cgtccccact ccgcgagacc ctggtgcacc tcaatcttgc tggttgccac
     3301 cgcctaacgg accactgcct cccgctgttc cgccgctgcc ctcgtctacg ccgcctagac
     3361 ctgcgctcct gccgccagct ctcacccgaa gcttgtgccc ggctggcagc tgccgggccc
     3421 cctggcccct tccgctgccc tgaggagaag ctgcttctca aggacagcta ggaattctgc
     3481 agatatccag cacagtggcg gccgctcgag tctagagggc ccgatatctc gacaatcaac
     3541 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3601 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3661 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3721 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3781 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3841 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3901 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3961 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4021 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4081 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct
     4141 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4201 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4261 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4321 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4381 gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg
     4441 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4501 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4561 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     4621 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4681 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4741 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4801 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4861 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4921 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5041 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5101 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     5161 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     5221 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     5281 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5341 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5401 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     5461 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5521 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5581 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5641 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5701 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5761 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5821 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5881 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5941 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6001 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6061 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     6121 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6181 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6241 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6301 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6361 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6421 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6481 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6541 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6601 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6661 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     7021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7501 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7561 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7621 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7681 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7741 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7801 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7861 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7921 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7981 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8041 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8101 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8161 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8221 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8281 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8341 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8401 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8581 cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
     8641 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8701 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
