LOCUS       sequence-426045-        7946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the K650M mutant variant of human FGFR3, which is 
            associated with Severe Achondroplasia with Developmental Delay and 
            Acanthosis Nigricans (SADDAN).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7946)
  AUTHORS   Kunova Bosakova M, Nita A, Gregor T, Varecha M, Gudernova I, Fafilek
            B, Barta T, Basheer N, Abraham SP, Balek L, Tomanova M, Fialova 
            Kucerova J, Bosak J, Potesil D, Zieba J, Song J, Konik P, Park S, 
            Duran I, Zdrahal Z, Smajs D, Jansen G, Fu Z, Ko HW, Hampl A, 
            Trantirek L, Krakow D, Krejci P
  TITLE     Fibroblast growth factor receptor influences primary cilium length 
            through an interaction with intestinal cell kinase.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Feb 19. pii: 1800338116. doi: 
            10.1073/pnas.1800338116.
  PUBMED    30782830
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: "Proc Natl 
            Acad Sci U S A. 2019 Feb 19. pii: 1800338116. doi: 
            10.1073/pnas.1800338116."
COMMENT     Sequence Label: pcDNA3.1/hFGFR3-K650M-V5
FEATURES             Location/Qualifiers
     source          1..7946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3444..3485
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3495..3512
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3535..3552)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3541..3765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3811..4239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3898..3917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4108..4129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4248..4268)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4253..4582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4433..4568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4495..4514
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4649..5443
                     /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(4703..4722)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5313..5332
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5619..5740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5656..5675)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5710..5729
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5789..5805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5789..5805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5802..5824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5813..5829
                     /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(5837..5843,5844..5861,5862..5867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5882..5903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6020..6037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6191..6779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6271..6290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6950..7741,7742..7810))
                     /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     7573..7592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7811..7915)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccagtac ccttcaccca cgaccccacc atgggcgccc
      961 ctgcctgcgc cctcgcgctc tgcgtggccg tggccatcgt ggccggcgcc tcctcggagt
     1021 ccttggggac ggagcagcgc gtcgtggggc gagcggcaga agtcccgggc ccagagcccg
     1081 gccagcagga gcagttggtc ttcggcagcg gggatgctgt ggagctgagc tgtcccccgc
     1141 ccgggggtgg tcccatgggg cccactgtct gggtcaagga tggcacaggg ctggtgccct
     1201 cggagcgtgt cctggtgggg ccccagcggc tgcaggtgct gaatgcctcc cacgaggact
     1261 ccggggccta cagctgccgg cagcggctca cgcagcgcgt actgtgccac ttcagtgtgc
     1321 gggtgacaga cgctccatcc tcgggagatg acgaagacgg ggaggacgag gctgaggaca
     1381 caggtgtgga cacaggggcc ccttactgga cacggcccga gcggatggac aagaagctgc
     1441 tggccgtgcc ggccgccaac accgtccgct tccgctgccc agccgctggc aaccccactc
     1501 cctccatctc ctggctgaag aacggcaggg agttccgcgg cgagcaccgc attggaggca
     1561 tcaagctgcg gcatcagcag tggagcctgg tcatggaaag cgtggtgccc tcggaccgcg
     1621 gcaactacac ctgcgtcgtg gagaacaagt ttggcagcat ccggcagacg tacacgctgg
     1681 acgtgctgga gcgctccccg caccggccca tcctgcaggc ggggctgccg gccaaccaga
     1741 cggcggtgct gggcagcgac gtggagttcc actgcaaggt gtacagtgac gcacagcccc
     1801 acatccagtg gctcaagcac gtggaggtga acggcagcaa ggtgggcccg gacggcacac
     1861 cctacgttac cgtgctcaag acggcgggcg ctaacaccac cgacaaggag ctagaggttc
     1921 tctccttgca caacgtcacc tttgaggacg ccggggagta cacctgcctg gcgggcaatt
     1981 ctattgggtt ttctcatcac tctgcgtggc tggtggtgct gccagccgag gaggagctgg
     2041 tggaggctga cgaggcgggc agtgtgtatg caggcatcct cagctacggg gtgggcttct
     2101 tcctgttcat cctggtggtg gcggctgtga cgctctgccg cctgcgcagc ccccccaaga
     2161 aaggcctggg ctcccccacc gtgcacaaga tctcccgctt cccgctcaag cgacaggtgt
     2221 ccctggagtc caacgcgtcc atgagctcca acacaccact ggtgcgcatc gcaaggctgt
     2281 cctcagggga gggccccacg ctggccaatg tctccgagct cgagctgcct gccgacccca
     2341 aatgggagct gtctcgggcc cggctgaccc tgggcaagcc ccttggggag ggctgcttcg
     2401 gccaggtggt catggcggag gccatcggca ttgacaagga ccgggccgcc aagcctgtca
     2461 ccgtagccgt gaagatgctg aaagacgatg ccactgacaa ggacctgtcg gacctggtgt
     2521 ctgagatgga gatgatgaag atgatcggga aacacaaaaa catcatcaac ctgctgggcg
     2581 cctgcacgca gggcgggccc ctgtacgtgc tggttgagta cgcggccaag ggtaacctgc
     2641 gggagtttct gcgggcgcgg cggcccccgg gcctggacta ctccttcgac acctgcaagc
     2701 cgcccgagga gcagctcacc ttcaaggacc tggtgtcctg tgcctaccag gtggcccggg
     2761 gcatggagta cttggcctcc cagaagtgca tccacaggga cctggctgcc cgcaatgtgc
     2821 tggtgaccga ggacaacgtg atgaagatcg cagacttcgg gctggcccgg gacgtgcaca
     2881 acctcgacta ctacaagatg acaaccaacg gccggctgcc cgtgaagtgg atggcgcctg
     2941 aggccttgtt tgaccgagtc tacactcacc agagtgacgt ctggtccttt ggggtcctgc
     3001 tctgggagat cttcacgctg gggggctccc cgtaccccgg catccctgtg gaggagctct
     3061 tcaagctgct gaaggagggc caccgcatgg acaagcccgc caactgcaca cacgacctgt
     3121 acatgatcat gcgggagtgc tggcatgccg cgccctccca gaggcccacc ttcaagcagc
     3181 tggtggagga cctggaccgt gtccttaccg tgacgtccac cgacgagtac ctggacctgt
     3241 cggcgccttt cgagcagtac tccccgggtg gccaggacac ccccagctcc agctcctcag
     3301 gggacgactc cgtgtttgcc cacgacctgc tgcccccggc cccacccagc agtgggggct
     3361 cgcggacggc caagggtcaa gacaattctg cagatatcca gcacagtggc ggccgctcga
     3421 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     3481 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     3541 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3601 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3661 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3721 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3781 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3841 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3901 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3961 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4021 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4081 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4141 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4201 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4261 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4321 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4381 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4441 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4501 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4561 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4621 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4681 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4741 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4801 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4861 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4921 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4981 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5041 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5101 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5161 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5221 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5281 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5341 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5401 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5461 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5521 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5581 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5641 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5701 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5761 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5821 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5881 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5941 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6001 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6061 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6121 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6601 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6781 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6841 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6901 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6961 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7021 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7081 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7141 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7201 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7261 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7321 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7381 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7441 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7501 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7561 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7621 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7681 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7741 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7801 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7861 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7921 ttccccgaaa agtgccacct gacgtc
//
