LOCUS       V025196                 8464 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025196
VERSION     V025196
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             1402..3138
                     /label="IGF2BP3(NM_006547)"
                     /note="IGF2BP3(NM_006547)"
                     /gene="IGF2BP3"
     CDS             3148..3213
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3240..3828
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3894..3942
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4144..4572
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4586..4915
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4982..5776
                     /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    5952..6073
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6122..6138)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6146..6162
                     /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(6170..6200)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6215..6236
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6524..7112)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7283..8143)
                     /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(8144..8248)
                     /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 ggcggtggag gttcaggtac catgaacaaa ctgtatatcg gaaacctcag cgagaacgcc
     1441 gccccctcgg acctagaaag tatcttcaag gacgccaaga tcccggtgtc gggacccttc
     1501 ctggtgaaga ctggctacgc gttcgtggac tgcccggacg agagctgggc cctcaaggcc
     1561 atcgaggcgc tttcaggtaa aatagaactg cacgggaaac ccatagaagt tgagcactcg
     1621 gtcccaaaaa ggcaaaggat tcggaaactt cagatacgaa atatcccgcc tcatttacag
     1681 tgggaggtgc tggatagttt actagtccag tatggagtgg tggagagctg tgagcaagtg
     1741 aacactgact cggaaactgc agttgtaaat gtaacctatt ccagtaagga ccaagctaga
     1801 caagcactag acaaactgaa tggatttcag ttagagaatt tcaccttgaa agtagcctat
     1861 atccctgatg aaatggccgc ccagcaaaac cccttgcagc agccccgagg tcgccggggg
     1921 cttgggcaga ggggctcctc aaggcagggg tctccaggat ccgtatccaa gcagaaacca
     1981 tgtgatttgc ctctgcgcct gctggttccc acccaatttg ttggagccat cataggaaaa
     2041 gaaggtgcca ccattcggaa catcaccaaa cagacccagt ctaaaatcga tgtccaccgt
     2101 aaagaaaatg cgggggctgc tgagaagtcg attactatcc tctctactcc tgaaggcacc
     2161 tctgcggctt gtaagtctat tctggagatt atgcataagg aagctcaaga tataaaattc
     2221 acagaagaga tccccttgaa gattttagct cataataact ttgttggacg tcttattggt
     2281 aaagaaggaa gaaatcttaa aaaaattgag caagacacag acactaaaat cacgatatct
     2341 ccattgcagg aattgacgct gtataatcca gaacgcacta ttacagttaa aggcaatgtt
     2401 gagacatgtg ccaaagctga ggaggagatc atgaagaaaa tcagggagtc ttatgaaaat
     2461 gatattgctt ctatgaatct tcaagcacat ttaattcctg gattaaatct gaacgccttg
     2521 ggtctgttcc cacccacttc agggatgcca cctcccacct cagggccccc ttcagccatg
     2581 actcctccct acccgcagtt tgagcaatca gaaacggaga ctgttcatct gtttatccca
     2641 gctctatcag tcggtgccat catcggcaag cagggccagc acatcaagca gctttctcgc
     2701 tttgctggag cttcaattaa gattgctcca gcggaagcac cagatgctaa agtgaggatg
     2761 gtgattatca ctggaccacc agaggctcag ttcaaggctc agggaagaat ttatggaaaa
     2821 attaaagaag aaaactttgt tagtcctaaa gaagaggtga aacttgaagc tcatatcaga
     2881 gtgccatcct ttgctgctgg cagagttatt ggaaaaggag gcaaaacggt gaatgaactt
     2941 cagaatttgt caagtgcaga agttgttgtc cctcgtgacc agacacctga tgagaatgac
     3001 caagtggttg tcaaaataac tggtcacttc tatgcttgcc aggttgccca gagaaaaatt
     3061 caggaaattc tgactcaggt aaagcagcac caacaacaga aggctctgca aagtggacca
     3121 cctcagtcaa gacggaaggc ctcgagagac tacaaagacc atgacggtga ttataaagat
     3181 catgacatcg actacaagga tgacgatgac aagtagtcta gagggcccga tatctcgaca
     3241 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3301 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3361 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3421 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3481 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3541 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3601 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3661 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3721 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3781 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     3841 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3901 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3961 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4021 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4081 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     4141 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4201 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4261 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     4321 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4381 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4441 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4501 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4561 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4621 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4681 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4741 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4801 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4861 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4921 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4981 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5041 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5101 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5161 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5221 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5281 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5341 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5401 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5461 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5521 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5581 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5641 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5701 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5761 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     5821 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5881 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5941 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6061 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6241 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     6721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6961 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8281 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     8341 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     8401 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     8461 tagg
//
