LOCUS       sequence-426031-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the mutant variant of CILK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6756)
  AUTHORS   Paige Taylor S, Kunova Bosakova M, Varecha M, Balek L, Barta T, 
            Trantirek L, Jelinkova I, Duran I, Vesela I, Forlenza KN, Martin JH,
            Hampl A, Bamshad M, Nickerson D, Jaworski ML, Song J, Ko HW, Cohn 
            DH, Krakow D, Krejci P
  TITLE     An inactivating mutation in intestinal cell kinase, ICK, impairs 
            hedgehog signalling and causes short rib-polydactyly syndrome.
  JOURNAL   Hum Mol Genet. 2016 Sep 15;25(18):3998-4011. doi: 
            10.1093/hmg/ddw240. Epub 2016 Jul 27.
  PUBMED    27466187
REFERENCE   2  (bases 1 to 6756)
  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"; doi: 
            "10.1093/hmg/ddw240"; journalName: "Hum Mol Genet"; date: 
            "2016-09-15- 15"; volume: "25"; issue: "18"; pages: "3998-4011"
COMMENT     Sequence Label: CILK1-E80K
FEATURES             Location/Qualifiers
     source          1..6756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1967..1975
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2943..2972
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2991..3014
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3034..3050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3034..3050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3061..3683
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3260..3279)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3832..4420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3912..3931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4749..4796
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4821..4840
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5028..5822)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5028..5822)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5139..5158)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5749..5768
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      5871..6005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6002..6193
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6150..6170
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6195..6299)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6326..6754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6413..6432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6623..6644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gaatagatac acaacaatca ggcagctcgg ggatggaacc tacggttccg
     1081 tcctgctggg aagaagcatt gagtctgggg agctgatcgc tattaaaaaa atgaaaagaa
     1141 aattttattc ctgggaggaa tgcatgaacc ttcgggaggt taagtcttta aagaagctca
     1201 accatgccaa tgtagtcaaa ttaaaagaag ttatcaggga aaatgatcat ctttatttta
     1261 tcttcaagta catgaaggaa aatctttacc agctcattaa agagagaaat aagttgtttc
     1321 ctgagtctgc tataaggaat atcatgtatc agatattaca aggactcgca tttattcaca
     1381 aacacggctt ctttcatcga gacttaaagc ctgagaacct cctctgcatg ggaccagaac
     1441 ttgtgaaaat tgcagacttt ggtttggccc gagaaatacg atcaaaacct ccatatacag
     1501 attatgtatc taccagatgg tacagggctc cagaagtact cctgaggtct accaactaca
     1561 gctcccccat tgacgtctgg gcggtgggct gcatcatggc agaagtttac accctcaggc
     1621 cactcttccc tggagccagt gaaattgaca caatattcaa aatttgccaa gtgctgggga
     1681 caccaaaaaa gactgactgg cctgaaggct atcaactttc aagtgcaatg aacttccgtt
     1741 ggccacagtg tgtacccaat aacttaaaga ccttgattcc caatgctagc agtgaagcag
     1801 tccagctcct gagagacatg cttcagtggg atcccaagaa acgaccaaca gctagtcagg
     1861 cacttcgata tccttacttc caagttggac acccactagg cagcaccaca caaaaccttc
     1921 aggattcaga aaaaccacag aaaggcatcc tggaaaaggc aggcccacct ccttatatta
     1981 agccagtccc acctgcccag ccaccagcca agccacacac acgaatttct tcacgacagc
     2041 atcaagccag ccagccccct ctgcatctca cgtaccccta caaagcagag gtctccagga
     2101 cagatcaccc aagccatctc caggaggaca agccaagccc gttgcttttc ccatccctcc
     2161 acaacaagca tccacagtcg aaaatcacag ctggcctgga gcacaaaaat ggtgagataa
     2221 agccaaagag taggagaagg tggggtctta tttccaggtc aacaaaggat tcagatgatt
     2281 gggctgactt ggatgacttg gatttcagtc catccctcag caggattgac ctgaaaaaca
     2341 agaaaagaca gagtgatgac actctctgca ggtttgagag tgttttggac ctgaagccct
     2401 ctgagcctgt gggcacagga aacagtgccc ccacccagac gtcatatcag cggcgagaca
     2461 cgcccaccct gagatctgca gccaagcagc actatttgaa gcactctcga tacttgcctg
     2521 ggatcagtat aagaaatggc atactctcga atccaggcaa ggaatttatt ccacctaatc
     2581 catggtctag ttctggcttg tctggaaaat cttcagggac aatgtcagta atcagcaaag
     2641 taaattcagt tggttccagc tctacaagtt ctagtggact gactggaaac tatgtccctt
     2701 cctttctgaa aaaagaaatc ggttctgcta tgcagagggt acacctagca cctattccag
     2761 acccttcccc tggttattcc tccctgaagg ccatgagacc tcatcctggg cgaccattct
     2821 tccacaccca gcctagaagc actcctgggt tgataccacg gcctccagcc gcccagccag
     2881 tgcatggccg gacagactgg gcttccaagt acgcatctcg gcgaacgcgt acgcggccgc
     2941 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg
     3001 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc
     3061 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3121 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3181 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3241 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3301 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     3361 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     3421 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     3481 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     3541 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     3601 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     3661 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc
     3721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4501 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4561 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     4621 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     4681 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     4741 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     4801 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     4861 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     4921 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     4981 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     5041 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     5101 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     5161 atgtcctgat agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     5221 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     5281 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     5341 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     5401 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     5461 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     5521 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     5581 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     5641 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     5701 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     5761 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     5821 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     5881 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     5941 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     6001 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     6061 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6121 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     6181 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6241 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6301 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6721 aatgagctga tttaacaaaa atttaacgcg aatttt
//
