LOCUS       sequence-348234-        6066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse ILK with R255A point mutation cloned into pEGFP-N1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6066)
  AUTHORS   Martin IM, Nava MM, Wickstrom SA, Grater F
  TITLE     ATP allosterically stabilizes integrin-linked kinase for efficient 
            force generation.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Mar 15;119(11):e2106098119. doi: 
            10.1073/pnas.2106098119. Epub 2022 Mar 8.
  PUBMED    35259013
REFERENCE   2  (bases 1 to 6066)
  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."; date: "2022-03-15"; volume: "119(11):e2106098119. 
            doi"; pages: " 10.1073/pnas.2106098119. Epub 2022 Mar 8"
COMMENT     Sequence Label: mILK-R255A-EFGP
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1642..1651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1648..1650,1651..1653,1654..2367)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1648..1650,1651..1653,1654..2364)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1693..1714)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1954..1973)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2301..2322
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2490..2611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2527..2546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2581..2600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2618..3073)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2755..2776)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2967..2986
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3100..3204
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3206..3563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3206..3397
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3229..3249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3414..3549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3598..4392
                     /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             3598..4392
                     /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(3652..3671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4262..4281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4580..4599)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4624..4671
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5000..5588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5489..5508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5763..6066
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc accatggacg acattttcac tcagtgccgg
      301 gagggcaacg cggtggcggt gcgcttgtgg ctggacaaca cagagaacga cctcaatcag
      361 ggggatgatc atggcttctc ccccttgcac tgggcctgcc gagaaggccg ctctgcggtg
      421 gttgaaatgc tgatcatgcg tggagcacgg attaatgtga tgaatcgtgg ggatgatacc
      481 cccctgcacc tggcagctag tcatggacac cgtgacattg tacagaagct gttgcaatac
      541 aaggctgaca tcaatgcagt gaatgagcac ggcaatgtgc cacttcacta tgcatgtttc
      601 tggggccaag accaggtggc agaggacctg gtggctaacg gggctcttgt gagcatctgt
      661 aacaagtatg gagagatgcc tgtggacaaa gccaaggcac cccttagaga gcttctccga
      721 gaacgggcag agaaaatggg ccagaatctc aaccgtattc catacaagga cacattctgg
      781 aaggggacca ctcgcacaag gccccgaaat gggaccctga acaaacactc cggtattgac
      841 ttcaaacagc tcaactttct ggcaaagctc aatgagaatc attctggaga gctttggaaa
      901 ggccgctggc agggcaatga tattgttgtg aaggtgctga aggttgcaga ctggagtaca
      961 aggaaaagca gggacttcaa tgaggaatgt ccccggctca ggattttctc acaccctaac
     1021 gtgctcccag tgctaggtgc ttgccaggct cccccagccc cccacccaac cctcatcaca
     1081 cactggatgc catatggatc tctctacaat gttctacatg aaggcaccaa tttcgttgtg
     1141 gaccagagcc aagctgtaaa gtttgctttg gacatggcaa gaggcatggc ttttcttcac
     1201 acactagagc ctctcatacc ccggcatgca ctcaatagcc gcagtgtaat gatcgatgaa
     1261 gatatgactg cccgaatcag catggctgat gttaagtttt ctttccagtg ccctgggcgc
     1321 atgtatgcgc ctgcctgggt ggcccctgaa gccctgcaga agaagcctga agacacaaac
     1381 agacgctcag cagacatgtg gagtttcgcg gtgcttctgt gggaactggt gacacgagag
     1441 gtgccctttg ctgacctctc taatatggag attgggatga aggtggcact ggaaggcctt
     1501 cggcctacca tcccaccagg tatttccccc catgtgtgta agctcatgaa gatttgcatg
     1561 aatgaagacc ctgcaaagcg acccaagttt gacatgattg tgcctatctt ggagaagatg
     1621 caggacaagg gaccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1681 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1741 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1801 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1861 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1921 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1981 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2041 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2101 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2161 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2221 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     2281 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2341 ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
     2401 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2461 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2581 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2641 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2701 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2761 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2821 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2881 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2941 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3001 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3061 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3181 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3241 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3481 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3541 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     3781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4381 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4441 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4501 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4561 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4621 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4681 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4741 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     4801 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     4861 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5641 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     5701 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     5761 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     5821 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     5881 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     5941 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6001 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6061 accatg
//
