LOCUS       pEF1-Hgf(rat)-3xFLAG    8828 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023645
VERSION     V023645
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        50..1228
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             1316..3499
                     /label="Hgf(NM_017017)"
                     /note="Hgf(NM_017017)"
                     /gene="Hgf"
     CDS             3515..3580
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3604..4192
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4258..4306
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4508..4936
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4950..5279
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5346..6140
                     /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    6316..6437
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6486..6502)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6510..6526
                     /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(6534..6564)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6579..6600
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6888..7476)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7647..8507)
                     /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(8508..8612)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg gctccggtgc
       61 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      121 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      181 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      241 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      301 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
      361 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      421 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      481 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      541 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      601 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      661 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      721 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      781 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
      841 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
      901 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
      961 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1021 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1081 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1141 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1201 tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactggcta gcgtttaaac
     1261 ttaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgat
     1321 gtgggggacc aaacttctgc cggtcctgtt gctgcagcat gtcctgctgc acctcctcct
     1381 gcttcctgtc accatcccct atgcagaagg acagaagaag agaagaaata ctcttcatga
     1441 attcaaaaag tcagcaaaaa ctactcttac caaggaagac ccattagtga agattaaaac
     1501 caaaaaagtg aactctgcag atgagtgtgc caacaggtgc atcagaaaca agggctttcc
     1561 attcacttgc aaggcctttg tttttgataa gtcgagaaaa cgatgctact ggtatccttt
     1621 caatagtatg tcaagtggag tgaaaaaagg gtttggccat gaatttgacc tctatgaaaa
     1681 caaagactat attagaaatt gcatcattgg taaaggaggc agctataagg ggacagtatc
     1741 catcactaag agtggcatca agtgccagcc ttggaattcc atgatccccc atgaacacag
     1801 ctttttgcct tcgagctatc gcggtaaaga cctacaggaa aactactgtc gaaatcctcg
     1861 aggggaagaa gggggaccct ggtgtttcac aagcaatcca gaggtacgct acgaagtctg
     1921 tgacattcct cagtgttcag aagttgaatg catgacctgc aacggtgaaa gctacagagg
     1981 tcccatggat cacacagaat caggcaagac atgtcagcgc tgggatcagc agacaccaca
     2041 ccggcacaaa ttcttgccgg aaagatatcc cgacaagggc tttgatgata attattgccg
     2101 caatcccgat ggcaagccga ggccatggtg ctacactctt gaccctgaca ccccttggga
     2161 gtattgtgca attaaaatgt gcgctcacag tgctgtgaat gagactgatg ttcccatgga
     2221 aacaactgaa tgtataaaag gccaaggaga aggttacagg ggaaccacca ataccatttg
     2281 gaatggaatt ccgtgtcagc gttgggattc gcagtacccc cacaagcatg acatcactcc
     2341 cgagaacttc aaatgcaagg accttagaga aaattattgc cgcaatccgg atggggctga
     2401 atcaccatgg tgttttacca ctgatccaaa catccgagtt ggttactgct ctcaaattcc
     2461 caaatgtgac gtgtcaagtg gacaagattg ttatcgtggc aatgggaaaa actacatggg
     2521 caacttatcc aaaacaaggt ctggactcac atgttccatg tgggacaaga atatggagga
     2581 tttacaccgt catatcttct gggagccaga cgctagcaag ttgactaaga attactgccg
     2641 gaaccccgat gacgacgccc atggaccttg gtgctacaca gggaatcctc tcgttccttg
     2701 ggattattgc cctatttccc gttgtgaagg agatactaca cctacaattg tcaatttgga
     2761 ccatcctgta atatcctgtg ccaaaacaaa acaactgcga gttgtaaatg gcattccaac
     2821 acaaacaaca gtagggtgga tggttagttt gaaatacagg aataaacaca tctgtggggg
     2881 atcattgata aaggaaagtt gggttcttac tgcaaggcaa tgttttccag ctagaaacaa
     2941 agacttgaaa gactatgaag cttggcttgg aatccatgat gtccatgaga gaggcgagga
     3001 gaaacgcaaa cagatcttaa acatttccca gctagtctat ggacctgaag gctcagattt
     3061 ggttttactg aagcttgctc gccctgcaat cctggataac tttgtcagta caattgattt
     3121 acctagttat ggctgtacaa tccctgaaaa gactacttgc agtatttacg gctggggcta
     3181 cactggattg atcaacgcag atggtttatt acgagtagct catctgtata ttatggggaa
     3241 tgagaaatgc agtcagcacc atcaaggcaa ggtgactttg aatgagtctg aattatgtgc
     3301 tggggctgaa aagattggat caggaccttg tgagggagat tatggtggcc cactcatttg
     3361 tgaacaacac aaaatgagaa tggttcttgg tgtcattgtt cctggtcgtg gatgtgccat
     3421 cccaaatcgt cctggtattt ttgttcgagt agcatattat gcaaaatgga tacacaaagt
     3481 aattttgaca tacaagttgc tgcggccgct cgaggactac aaagaccatg acggtgatta
     3541 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     3601 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3661 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3721 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3781 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3841 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3901 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3961 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4021 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4081 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4141 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     4201 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4261 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     4321 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4381 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4441 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     4501 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4561 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4621 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     4681 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4741 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4801 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4861 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4921 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4981 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5101 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5161 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5281 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5341 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5401 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5461 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5521 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5581 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5641 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5701 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5761 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5821 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5881 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5941 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6001 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6061 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6121 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6181 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6241 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6301 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6361 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6421 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6481 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6541 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6601 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6661 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6721 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6781 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7081 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7321 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8641 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     8701 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8761 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8821 tgcttagg
//
