LOCUS       sequence-394496-        7976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human FGFR1 receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  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 7976)
  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-FGFR1c-V5/HIS
FEATURES             Location/Qualifiers
     source          1..7976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..45)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        217..596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        597..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     751..771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     845..864
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        845..863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1300..1317)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3456..3497
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3507..3524
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3547..3564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3553..3777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3823..4251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3910..3929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4120..4141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4260..4280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4265..4594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4445..4580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4507..4526
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4661..5455
                     /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(4715..4734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5325..5344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5631..5752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5668..5687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5722..5741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5801..5817)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5801..5817)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5814..5836)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5825..5841
                     /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(5849..5855,5856..5873,5874..5879))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5894..5915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6032..6049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6203..6791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6283..6302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6962..7753,7754..7822))
                     /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     7585..7604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7823..7927)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
       61 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
      121 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      181 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      241 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      781 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
      841 aaattaatac gactcactat agggagaccc aagctggcta gttaagcttg gtaccgagct
      901 cggatccagt acccttcacc atgtggagat ggaagtgcct cctcttctgg gctgtgctgg
      961 tcacagccac actctgcacc gctaggccgt ccccgacctt gcctgaacaa gcccagccct
     1021 ggggagcccc tgtggaagtg gagtccttcc tggtccaccc cggtgacctg ctgcagcttc
     1081 gctgtcggct gcgggacgat gtgcagagca tcaactggct gcgggacggg gtgcagctgg
     1141 cggaaagcaa ccgcacccgc atcacagggg aggaggtgga ggtgcaggac tccgtgcccg
     1201 cagactccgg cctctatgct tgcgtaacca gcagcccctc gggcagtgac accacctact
     1261 tctccgtcaa tgtttcagat gctctcccct cctcggagga tgatgatgat gatgatgact
     1321 cctcttcaga ggagaaagaa acagataaca ccaaaccaaa ccccgtagct ccatattgga
     1381 catccccaga aaagatggaa aagaaattgc atgcagtgcc ggctgccaag acagtgaagt
     1441 tcaaatgccc ttccagtggg accccaaacc ccacactgcg ctggttgaaa aatggcaaag
     1501 aattcaaacc tgaccacaga attggaggct acaaggtccg ttatgccacc tggagcatca
     1561 taatggactc tgtggtgccc tctgacaagg gcaactacac ctgcattgtg gagaatgagt
     1621 acggcagcat caaccacaca taccagctgg atgtcgtgga gcggtcccct caccggccca
     1681 tcctgcaagc agggttgccc gccaacaaaa cagtggccct gggtagcaac gtggagttca
     1741 tgtgtaaggt gtacagtgac ccgcagccgc acatccagtg gctaaagcac atcgaggtga
     1801 atgggagcaa gattggccca gacaacctgc cttatgtcca gatcttgaag actgctggag
     1861 ttaataccac cgacaaagag atggaggtgc ttcacttaag aaatgtctcc tttgaggacg
     1921 caggggagta tacgtgcttg gcgggtaact ctatcggact ctcccatcac tctgcatggt
     1981 tgaccgttct ggaagccctg gaagagaggc cggcagtgat gacctcgccc ctgtacctgg
     2041 agatcatcat ctattgcaca ggggccttcc tcatctcctg catggtgggg tcggtcatcg
     2101 tctacaagat gaagagtggt accaagaaga gtgacttcca cagccagatg gctgtgcaca
     2161 agctggccaa gagcatccct ctgcgcagac aggtaacagt gtctgctgac tccagtgcat
     2221 ccatgaactc tggggttctt ctggttcggc catcacggct ctcctccagt gggactccca
     2281 tgctagcagg ggtctctgag tatgagcttc ccgaagaccc tcgctgggag ctgcctcggg
     2341 acagactggt cttaggcaaa cccctgggag agggctgctt tgggcaggtg gtgttggcag
     2401 aggctatcgg gctggacaag gacaaaccca accgtgtgac caaagtggct gtgaagatgt
     2461 tgaagtcgga cgcaacagag aaagacttgt cagacctgat ctcagaaatg gagatgatga
     2521 agatgatcgg gaagcataag aatatcatca acctgctggg ggcctgcacg caggatggtc
     2581 ccttgtatgt catcgtggag tatgcctcca agggcaacct gcgggagtac ctgcaggccc
     2641 ggaggccccc agggctggaa tactgctaca accccagcca caacccagag gagcagctct
     2701 cctccaagga cctggtgtcc tgcgcctacc aggtggcccg aggcatggag tatctggcct
     2761 ccaagaagtg catacaccga gacctggcag ccaggaatgt cctggtgaca gaggacaatg
     2821 tgatgaagat agcagacttt ggcctcgcac gggacattca ccacatcgac tactataaaa
     2881 agacaaccaa cggccgactg cctgtgaagt ggatggcacc cgaggcatta tttgaccgga
     2941 tctacaccca ccagagtgat gtgtggtctt tcggggtgct cctgtgggag atcttcactc
     3001 tgggcggctc cccatacccc ggtgtgcctg tggaggaact tttcaagctg ctgaaggagg
     3061 gtcaccgcat ggacaagccc agtaactgca ccaacgagct gtacatgatg atgcgggact
     3121 gctggcatgc agtgccctca cagagaccca ccttcaagca gctggtggaa gacctggacc
     3181 gcatcgtggc cttgacctcc aaccaggagt acctggacct gtccatgccc ctggaccagt
     3241 actcccccag ctttcccgac acccggagct ctacgtgctc ctcaggggag gattccgtct
     3301 tctctcatga gccgctgccc gaggagccct gcctgccccg acacccagcc cagcttgcca
     3361 atggcggact caaacgccgc gccaagggtc aagacaattc tgcagatatc cagcacagtg
     3421 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     3481 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     3541 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3601 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3661 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3721 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3781 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3841 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3901 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3961 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4021 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4081 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4141 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4201 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4261 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4321 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4381 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4441 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4501 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4561 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4621 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4681 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4741 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4801 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4861 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4921 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4981 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5041 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5101 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5161 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5221 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5281 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5341 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5401 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5461 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5521 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5581 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5641 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5701 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5761 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5821 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5881 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5941 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6001 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6061 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6121 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6181 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6241 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6301 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6361 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6421 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6481 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6541 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6601 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6661 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6721 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6961 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7021 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7081 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7141 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7201 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7261 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7321 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7381 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7441 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7501 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7561 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7621 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7681 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7741 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7801 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7861 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7921 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctc
//
