LOCUS       sequence-348237-        6066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse ILK with R349A 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-R349A-EFGP
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1946..1955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1952..1954,1955..1957,1958..2671)
                     /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(1952..1954,1955..1957,1958..2668)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1997..2018)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2258..2277)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2605..2626
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2794..2915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2831..2850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2885..2904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2922..3377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3059..3080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3271..3290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3404..3508
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3510..3867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3510..3701
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3533..3553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3718..3853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3902..4696
                     /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             3902..4696
                     /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(3956..3975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4566..4585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4884..4903)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4928..4975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5304..5892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5793..5812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcaccatg gacgacattt tcactcagtg
      601 ccgggagggc aacgcggtgg cggtgcgctt gtggctggac aacacagaga acgacctcaa
      661 tcagggggat gatcatggct tctccccctt gcactgggcc tgccgagaag gccgctctgc
      721 ggtggttgaa atgctgatca tgcgtggagc acggattaat gtgatgaatc gtggggatga
      781 tacccccctg cacctggcag ctagtcatgg acaccgtgac attgtacaga agctgttgca
      841 atacaaggct gacatcaatg cagtgaatga gcacggcaat gtgccacttc actatgcatg
      901 tttctggggc caagaccagg tggcagagga cctggtggct aacggggctc ttgtgagcat
      961 ctgtaacaag tatggagaga tgcctgtgga caaagccaag gcacccctta gagagcttct
     1021 ccgagaacgg gcagagaaaa tgggccagaa tctcaaccgt attccataca aggacacatt
     1081 ctggaagggg accactcgca caaggccccg aaatgggacc ctgaacaaac actccggtat
     1141 tgacttcaaa cagctcaact ttctggcaaa gctcaatgag aatcattctg gagagctttg
     1201 gaaaggccgc tggcagggca atgatattgt tgtgaaggtg ctgaaggttc gagactggag
     1261 tacaaggaaa agcagggact tcaatgagga atgtccccgg ctcaggattt tctcacaccc
     1321 taacgtgctc ccagtgctag gtgcttgcca ggctccccca gccccccacc caaccctcat
     1381 cacacactgg atgccatatg gatctctcta caatgttcta catgaaggca ccaatttcgt
     1441 tgtggaccag agccaagctg taaagtttgc tttggacatg gcaagaggca tggcttttct
     1501 tcacacacta gagcctctca taccccggca tgcactcaat agccgcagtg taatgatcga
     1561 tgaagatatg actgcccgaa tcagcatggc tgatgttaag ttttctttcc agtgccctgg
     1621 ggccatgtat gcgcctgcct gggtggcccc tgaagccctg cagaagaagc ctgaagacac
     1681 aaacagacgc tcagcagaca tgtggagttt cgcggtgctt ctgtgggaac tggtgacacg
     1741 agaggtgccc tttgctgacc tctctaatat ggagattggg atgaaggtgg cactggaagg
     1801 ccttcggcct accatcccac caggtatttc cccccatgtg tgtaagctca tgaagatttg
     1861 catgaatgaa gaccctgcaa agcgacccaa gtttgacatg attgtgccta tcttggagaa
     1921 gatgcaggac aagggaccac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1981 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2041 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2101 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2161 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2221 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2281 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2341 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2401 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2461 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2521 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2581 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2641 cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
     2701 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2761 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2821 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2881 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2941 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3001 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3061 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3121 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3181 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3241 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3301 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3361 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3481 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3541 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3601 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3661 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3841 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3901 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3961 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4021 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4081 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4141 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4201 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4261 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4321 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4381 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4441 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4501 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4561 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4621 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4681 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4741 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4801 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4861 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4921 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4981 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5041 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5101 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5161 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5221 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5281 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5341 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5401 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5461 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5521 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5581 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5641 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5701 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5761 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5821 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5881 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5941 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6001 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6061 gttccg
//
