LOCUS       sequence-394493-        7946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human FGFR3 receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7946)
  AUTHORS   Gudernova I, Foldynova-Trantirkova S, Ghannamova BE, Fafilek B, 
            Varecha M, Balek L, Hruba E, Jonatova L, Jelinkova I, Kunova 
            Bosakova M, Trantirek L, Mayer J, Krejci P
  TITLE     One reporter for in-cell activity profiling of majority of protein 
            kinase oncogenes.
  JOURNAL   Elife. 2017 Feb 15;6. pii: e21536. doi: 10.7554/eLife.21536.
  PUBMED    28199182
REFERENCE   2  (bases 1 to 7946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2017-02-15"; volume: "6. pii: e21536. doi"; pages: " 
            10.7554/eLife.21536"
COMMENT     Sequence Label: pcDNA3.1-FGFR3c-V5/HIS
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3432..3473
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3483..3500
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3523..3540)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3529..3753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3799..4227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3886..3905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4096..4117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4236..4256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4241..4570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4421..4556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4483..4502
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4637..5431
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4691..4710)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5301..5320
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5607..5728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5644..5663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5698..5717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5777..5793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5777..5793)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5790..5812)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5801..5817
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5825..5831,5832..5849,5850..5855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5870..5891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6008..6025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6179..6767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6259..6278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6938..7729,7730..7798))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7561..7580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7799..7903)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      841 ttatcgaaat taatacgact cactataggg agacccaagc tggctagtta agcttggtac
      901 cgagctcgga tccagtaccc ttcacccacg accccaccat gggcgcccct gcctgcgccc
      961 tcgcgctctg cgtggccgtg gccatcgtgg ccggcgcctc ctcggagtcc ttggggacgg
     1021 agcagcgcgt cgtggggcga gcggcagaag tcccgggccc agagcccggc cagcaggagc
     1081 agttggtctt cggcagcggg gatgctgtgg agctgagctg tcccccgccc gggggtggtc
     1141 ccatggggcc cactgtctgg gtcaaggatg gcacagggct ggtgccctcg gagcgtgtcc
     1201 tggtggggcc ccagcggctg caggtgctga atgcctccca cgaggactcc ggggcctaca
     1261 gctgccggca gcggctcacg cagcgcgtac tgtgccactt cagtgtgcgg gtgacagacg
     1321 ctccatcctc gggagatgac gaagacgggg aggacgaggc tgaggacaca ggtgtggaca
     1381 caggggcccc ttactggaca cggcccgagc ggatggacaa gaagctgctg gccgtgccgg
     1441 ccgccaacac cgtccgcttc cgctgcccag ccgctggcaa ccccactccc tccatctcct
     1501 ggctgaagaa cggcagggag ttccgcggcg agcaccgcat tggaggcatc aagctgcggc
     1561 atcagcagtg gagcctggtc atggaaagcg tggtgccctc ggaccgcggc aactacacct
     1621 gcgtcgtgga gaacaagttt ggcagcatcc ggcagacgta cacgctggac gtgctggagc
     1681 gctccccgca ccggcccatc ctgcaggcgg ggctgccggc caaccagacg gcggtgctgg
     1741 gcagcgacgt ggagttccac tgcaaggtgt acagtgacgc acagccccac atccagtggc
     1801 tcaagcacgt ggaggtgaac ggcagcaagg tgggcccgga cggcacaccc tacgttaccg
     1861 tgctcaagac ggcgggcgct aacaccaccg acaaggagct agaggttctc tccttgcaca
     1921 acgtcacctt tgaggacgcc ggggagtaca cctgcctggc gggcaattct attgggtttt
     1981 ctcatcactc tgcgtggctg gtggtgctgc cagccgagga ggagctggtg gaggctgacg
     2041 aggcgggcag tgtgtatgca ggcatcctca gctacggggt gggcttcttc ctgttcatcc
     2101 tggtggtggc ggctgtgacg ctctgccgcc tgcgcagccc ccccaagaaa ggcctgggct
     2161 cccccaccgt gcacaagatc tcccgcttcc cgctcaagcg acaggtgtcc ctggagtcca
     2221 acgcgtccat gagctccaac acaccactgg tgcgcatcgc aaggctgtcc tcaggggagg
     2281 gccccacgct ggccaatgtc tccgagctcg agctgcctgc cgaccccaaa tgggagctgt
     2341 ctcgggcccg gctgaccctg ggcaagcccc ttggggaggg ctgcttcggc caggtggtca
     2401 tggcggaggc catcggcatt gacaaggacc gggccgccaa gcctgtcacc gtagccgtga
     2461 agatgctgaa agacgatgcc actgacaagg acctgtcgga cctggtgtct gagatggaga
     2521 tgatgaagat gatcgggaaa cacaaaaaca tcatcaacct gctgggcgcc tgcacgcagg
     2581 gcgggcccct gtacgtgctg gttgagtacg cggccaaggg taacctgcgg gagtttctgc
     2641 gggcgcggcg gcccccgggc ctggactact ccttcgacac ctgcaagccg cccgaggagc
     2701 agctcacctt caaggacctg gtgtcctgtg cctaccaggt ggcccggggc atggagtact
     2761 tggcctccca gaagtgcatc cacagggacc tggctgcccg caatgtgctg gtgaccgagg
     2821 acaacgtgat gaagatcgca gacttcgggc tggcccggga cgtgcacaac ctcgactact
     2881 acaagaagac aaccaacggc cggctgcccg tgaagtggat ggcgcctgag gccttgtttg
     2941 accgagtcta cactcaccag agtgacgtct ggtcctttgg ggtcctgctc tgggagatct
     3001 tcacgctggg gggctccccg taccccggca tccctgtgga ggagctcttc aagctgctga
     3061 aggagggcca ccgcatggac aagcccgcca actgcacaca cgacctgtac atgatcatgc
     3121 gggagtgctg gcatgccgcg ccctcccaga ggcccacctt caagcagctg gtggaggacc
     3181 tggaccgtgt ccttaccgtg acgtccaccg acgagtacct ggacctgtcg gcgcctttcg
     3241 agcagtactc cccgggtggc caggacaccc ccagctccag ctcctcaggg gacgactccg
     3301 tgtttgccca cgacctgctg cccccggccc cacccagcag tgggggctcg cggacggcca
     3361 agggtcaaga caattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc
     3421 cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     3481 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     3541 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3601 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3661 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3721 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3781 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3841 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3901 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3961 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4021 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4081 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4141 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4201 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4321 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4381 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4441 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4501 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4561 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4621 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4681 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4741 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4801 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4861 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4921 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4981 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5041 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5101 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5161 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5221 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5281 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5341 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5401 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     5461 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5521 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5581 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5641 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5701 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5761 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5821 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5881 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5941 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6001 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6061 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6121 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6181 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6241 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6301 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6361 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6421 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6481 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6541 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6601 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6661 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6721 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6781 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6841 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6901 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6961 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7021 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7081 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7141 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7201 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7261 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7321 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7381 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7441 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7501 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7561 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7621 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7681 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7741 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7801 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7861 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7921 tgccacctga cgtcgacgga tcggga
//
