LOCUS       sequence-394499-        7979 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expression of human FGFR2 receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  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 7979)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-FGFR2c-V5/HIS
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..46)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        218..597
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..801
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     752..772
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     846..865
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        846..864
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1318..1329
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             3460..3501
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3511..3528
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3551..3568)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3557..3781
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3827..4255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3914..3933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4124..4145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4264..4284)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4269..4598
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4449..4584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4511..4530
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4665..5459
                     /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="MIEQDGLHAGSPAA*VERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4719..4738)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5329..5348
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5635..5756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5672..5691)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5726..5745
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5805..5821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5805..5821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5818..5840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5829..5845
                     /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(5853..5859,5860..5877,5878..5883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5898..5919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6036..6053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6207..6795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6287..6306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6966..7757,7758..7826))
                     /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     7589..7608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7827..7931)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
       61 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
      121 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
      181 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      241 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      301 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      361 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      421 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      481 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      541 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      601 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      661 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      721 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      781 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
      841 gaaattaata cgactcacta tagggagacc caagctggct agttaagctt ggtaccgagc
      901 tcggatccag tacccttcac catggtcagc tggggtcgtt tcatctgcct ggtcgtggtc
      961 accatggcaa ccttgtccct ggcccggccc tccttcagtt tagttgagga taccacatta
     1021 gagccagaag agccaccaac caaataccaa atctctcaac cagaagtgta cgtggctgcg
     1081 ccaggggagt cgctagaggt gcgctgcctg ttgaaagatg ccgccgtgat cagttggact
     1141 aaggatgggg tgcacttggg gcccaacaat aggacagtgc ttattgggga gtacttgcag
     1201 ataaagggcg ccacgcctag agactccggc ctctatgctt gtactgccag taggactgta
     1261 gacagtgaaa cttggtactt catggtgaat gtcacagatg ccatctcatc cggagatgat
     1321 gaggatgaca ccgatggtgc ggaagatttt gtcagtgaga acagtaacaa caagagagca
     1381 ccatactgga ccaacacaga aaagatggaa aagcggctcc atgctgtgcc tgcggccaac
     1441 actgtcaagt ttcgctgccc agccggggga aacccaatgc caaccatgcg gtggctgaaa
     1501 aacgggaagg agtttaagca ggagcatcgc attggaggct acaaggtacg aaaccagcac
     1561 tggagcctca ttatggaaag tgtggtccca tctgacaagg gaaattatac ctgtgtggtg
     1621 gagaatgaat acgggtccat caatcacacg taccacctgg atgttgtgga gcgatcgcct
     1681 caccggccca tcctccaagc cggactgccg gcaaatgcct ccacagtggt cggaggagac
     1741 gtagagtttg tctgcaaggt ttacagtgat gcccagcccc acatccagtg gatcaagcac
     1801 gtggaaaaga acggcagtaa atacgggccc gacgggctgc cctacctcaa ggttctcaag
     1861 gccgccggtg ttaacaccac ggacaaagag attgaggttc tctatattcg gaatgtaact
     1921 tttgaggacg ctggggaata tacgtgcttg gcgggtaatt ctattgggat atcctttcac
     1981 tctgcatggt tgacagttct gccagcgcct ggaagagaaa aggagattac agcttcccca
     2041 gactacctgg agatagccat ttactgcata ggggtcttct taatcgcctg tatggtggta
     2101 acagtcatcc tgtgccgaat gaagaacacg accaagaagc cagacttcag cagccagccg
     2161 gctgtgcaca agctgaccaa acgtatcccc ctgcggagac aggtaacagt ttcggctgag
     2221 tccagctcct ccatgaactc caacaccccg ctggtgagga taacaacacg cctctcttca
     2281 acggcagaca cccccatgct ggcaggggtc tccgagtatg aacttccaga ggacccaaaa
     2341 tgggagtttc caagagataa gctgacactg ggcaagcccc tgggagaagg ttgctttggg
     2401 caagtggtca tggcggaagc agtgggaatt gacaaagaca agcccaagga ggcggtcacc
     2461 gtggccgtga agatgttgaa agatgatgcc acagagaaag acctttctga tctggtgtca
     2521 gagatggaga tgatgaagat gattgggaaa cacaagaata tcataaatct tcttggagcc
     2581 tgcacacagg atgggcctct ctatgtcata gttgagtatg cctctaaagg caacctccga
     2641 gaatacctcc gagcccggag gccacccggg atggagtact cctatgacat taaccgtgtt
     2701 cctgaggagc agatgacctt caaggacttg gtgtcatgca cctaccagct ggccagaggc
     2761 atggagtact tggcttccca aaaatgtatt catcgagatt tagcagccag aaatgttttg
     2821 gtaacagaaa acaatgtgat gaaaatagca gactttggac tcgccagaga tatcaacaat
     2881 atagactatt acaaaaagac caccaatggg cggcttccag tcaagtggat ggctccagaa
     2941 gccctgtttg atagagtata cactcatcag agtgatgtct ggtccttcgg ggtgttaatg
     3001 tgggagatct tcactttagg gggctcgccc tacccaggga ttcccgtgga ggaacttttt
     3061 aagctgctga aggaaggaca cagaatggat aagccagcca actgcaccaa cgaactgtac
     3121 atgatgatga gggactgttg gcatgcagtg ccctcccaga gaccaacgtt caagcagttg
     3181 gtagaagact tggatcgaat cctcactctc acaaccaatg aggaatactt ggacctcagc
     3241 caacctctcg aacagtattc acctagttac cctgacacaa gaagttcttg ttcttcagga
     3301 gatgattctg ttttttctcc agaccccatg ccttacgaac catgccttcc tcagtatcca
     3361 cacataaacg gcagtgttaa aacagcgaag ggtcaagaca attctgcaga tatccagcac
     3421 agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct
     3481 ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc
     3541 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3601 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3661 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3721 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3781 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3841 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3901 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3961 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4021 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4081 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4141 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4201 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4261 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4321 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4441 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4501 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4561 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4621 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4681 gattgcacgc aggttctccg gccgcttagg tggagaggct attcggctat gactgggcac
     4741 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4801 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4861 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4921 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4981 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5041 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5101 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5161 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5221 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5281 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5341 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5401 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5461 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5521 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5581 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5641 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5701 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5761 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5821 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5881 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5941 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6001 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7561 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatct
//
