LOCUS       sequence-209777-        8824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses FGFR2 with a single amino acid change in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8824)
  TITLE     FGFR2 unpublished mutants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8824)
  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: 
            "Unpublished"
COMMENT     Sequence Label: FGFR2 N549K 3xFlag
FEATURES             Location/Qualifiers
     source          1..8824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..975
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     protein_bind    1429..1440
                     /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             3508..3573
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3584..4206
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3783..3802)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4230..4250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4248..4545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4599..5393
                     /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(4653..4672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5263..5282
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6045..6179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6082..6101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6136..6155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6184..6203)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6185..6203)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6217..6233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6217..6233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6230..6252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6241..6257
                     /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(6265..6271,6272..6289,6290..6295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6447..6464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6618..7206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6698..6717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7377..8168,8169..8237))
                     /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     8000..8019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8238..8342)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8369..8797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8456..8475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8666..8687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attacggacc atgtctgcac ttctgatcct agctcttgtt
      961 ggagctgcag ttgctaagct tgcggccgcg aattcatcga tagatctgat atcggtacca
     1021 gtcgactcta gaatggtcag ctggggtcgt ttcatctgcc tggtcgtggt caccatggca
     1081 accttgtccc tggcccggcc ctccttcagt ttagttgagg ataccacatt agagccagaa
     1141 gagccaccaa ccaaatacca aatctctcaa ccagaagtgt acgtggctgc gccaggggag
     1201 tcgctagagg tgcgctgcct gttgaaagat gccgccgtga tcagttggac taaggatggg
     1261 gtgcacttgg ggcccaacaa taggacagtg cttattgggg agtacttgca gataaagggc
     1321 gccacgccta gagactccgg cctctatgct tgtactgcca gtaggactgt agacagtgaa
     1381 acttggtact tcatggtgaa tgtcacagat gccatctcat ccggagatga tgaggatgac
     1441 accgacggtg cggaagattt tgtcagtgag aacagtaaca acaagagagc accatactgg
     1501 accaacacag aaaagatgga aaagcggctc catgctgtgc ctgcggccaa cactgtcaag
     1561 tttcgctgcc cagccggggg gaacccaatg ccaaccatgc ggtggctgaa aaacgggaag
     1621 gagtttaagc aggagcatcg cattggaggc tacaaggtac gaaaccagca ctggagcctc
     1681 attatggaaa gtgtggtccc atctgacaag ggaaattata cctgtgtggt ggagaatgaa
     1741 tacgggtcca tcaatcacac gtaccacctg gatgttgtgg agcgatcgcc tcaccggccc
     1801 atcctccaag ccggactgcc ggcaaatgcc tccacagtgg ttggaggaga cgtagagttt
     1861 gtctgcaagg tttacagtga tgcccagccc cacatccagt ggatcaagca cgtggaaaag
     1921 aacggcagta aatacgggcc cgacgggctg ccctacctca aggttctcaa ggccgccggt
     1981 gttaacacca cggacaaaga gattgaggtt ctctatattc ggaatgtaac ttttgaggac
     2041 gctggggaat atacgtgctt ggcgggtaat tctattggga tatcctttca ctctgcatgg
     2101 ttgacagttc tgccagcgcc tggaagagaa aaggagatta cagcttcccc agactacctg
     2161 gagatagcca tttactgcat aggggtcttc ttaatcgcct gtatggtggt aacagtcatc
     2221 ctgtgccgaa tgaagaacac gaccaagaag ccagacttca gcagccagcc ggctgtgcac
     2281 aagctgacca aacgtatccc cctgcggaga caagtaacag tttcggctga gtccagctcc
     2341 tccatgaact ccaacacccc gctggtgagg ataacaacac gcctctcttc aacggcagac
     2401 acccccatgc tggcaggggt ctccgagtat gaacttccag aggacccaaa atgggagttt
     2461 ccaagagata agctgacact gggcaagccc ctgggagaag gttgctttgg gcaagtggtc
     2521 atggcggaag cagtgggaat tgacaaagac aagcccaagg aggcggtcac cgtggccgtg
     2581 aagatgttga aagatgatgc cacagagaaa gacctttctg atctggtgtc agagatggag
     2641 atgatgaaga tgattgggaa acacaagaat atcataaagc ttcttggagc ctgcacacag
     2701 gatgggcctc tctatgtcat agttgagtat gcctctaaag gcaacctccg agaatacctc
     2761 cgagcccgga ggccacccgg gatggagtac tcctatgaca ttaaccgtgt tcctgaggag
     2821 cagatgacct tcaaggactt ggtgtcatgc acctaccagc tggccagagg catggagtac
     2881 ttggcttccc aaaaatgtat tcatcgagat ttagcagcca gaaatgtttt ggtaacagaa
     2941 aacaatgtga tgaaaatagc agactttgga ctcgccagag atatcaacaa tatagactat
     3001 tacaaaaaga ccaccaatgg gcggcttcca gtcaagtgga tggctccaga agccctgttt
     3061 gatagagtat acactcatca gagtgatgtc tggtccttcg gggtgttaat gtgggagatc
     3121 ttcactttag ggggctcgcc ctacccaggg attcccgtgg aggaactttt taagctgctg
     3181 aaggaaggac acagaatgga taagccagcc aactgcacca acgaactgta catgatgatg
     3241 agggactgtt ggcatgcagt gccctcccag agaccaacgt tcaagcagtt ggtagaagac
     3301 ttggatcgaa ttctcactct cacaaccaat gaggaatact tggacctcag ccaacctctc
     3361 gaacagtatt cacctagtta ccctgacaca agaagttctt gttcttcagg agatgattct
     3421 gttttttctc cagaccccat gccttacgaa ccatgccttc ctcagtatcc acacataaac
     3481 ggcagtgtta aaacaggatc ccgggctgac tacaaagacc atgacggtga ttataaagat
     3541 catgacatcg actacaagga tgacgatgac aagtagtgat cccgggtggc atccctgtga
     3601 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     3661 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     3721 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3781 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3841 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3901 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3961 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4021 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata
     4081 ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca
     4141 tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt
     4201 tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4501 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggaa ttgatcagct
     4561 tgggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4741 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5101 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5401 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5461 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5521 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct
     5581 cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg
     5641 caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact
     5701 gcaggagtgg ggaggcacga tggccgcttt ggtcgacccg gacgggacgc tcctgcgcct
     5761 gatacagaac gaattgcttg caggcatctc atgagtgtgt cttcccgttt tccgcctgag
     5821 gtcactgcgt ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct caccacccac
     5881 tcgccaagct ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgccga
     5941 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     6001 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     6061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     6181 attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     6241 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6301 gggtgcctaa tgagtggcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6361 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6421 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6481 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6541 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7021 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7441 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7501 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7561 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7621 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7681 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7741 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7801 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7861 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7921 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7981 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8041 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8101 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8161 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8221 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8341 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     8401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8521 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8701 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     8761 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     8821 attt
//
