LOCUS       sequence-358840-        5654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of human paxillin LIM domains 1, 3, 3 and 4 with 
            N-terminal GST fusion protein in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Ripamonti M, Liaudet N, Azizi L, Bouvard D, Hytonen VP, 
            Wehrle-Haller B
  TITLE     Structural and functional analysis of LIM domain-dependent 
            recruitment of paxillin to alphavbeta3 integrin-positive focal 
            adhesions.
  JOURNAL   Commun Biol. 2021 Mar 29;4(1):380. doi: 10.1038/s42003-021-01886-9.
  PUBMED    33782527
REFERENCE   2  (bases 1 to 5654)
  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: "Commun 
            Biol."; date: "2021-03-29"; volume: "4(1):380. doi"; pages: " 
            10.1038/s42003-021-01886-9"
COMMENT     Sequence Label: GST_LIM1334
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1704..1726)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1871..1889)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1957..2061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2062..2130,2131..2922)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2280..2299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3093..3681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3582..3601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3835..3852
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3925..4002
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4003..5085
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4022..4041)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5134..5139,5140..5157,5158..5164)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5172..5188
                     /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)."
     primer_bind     5196..5212
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5196..5212
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5208..5654,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5228..5245)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5228..5244)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5237..5259)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatctggagt ctgcggggcc tgcaagaagc
      961 ccatcgccgg gcaggttgtg accgccatgg ggaagacgtg gcaccccgag cacttcgtct
     1021 gcacccactg ccaggaggag atcggatccc ggaacttctt cgagcgggat ggacagccct
     1081 actgtgaaaa ggactaccac aacctcttct cccccaagtg cggcgggtgt gcccgggcta
     1141 tccttgaaaa ctatatttca gccctgaata cgctctggca tcctgagtgt ttcgtctgcc
     1201 gcgaatgttt cactccattt gtgaatggga gcttctttga gcacgatggc cagccttact
     1261 gcgaagtcca ttatcacgag cggcgcgcac ctaaatgtgg gggctgcgct cgcgccattc
     1321 tggagaatta catctccgct ctcaacactc tgtggcaccc cgaatgcttt gtgtgtcggg
     1381 agtgctttac gcctttcgtc aacggcagtt ttttcgaaca tgacgggcag ccctattgtg
     1441 aggtgcacta ccatgaacgt cggggctcgc tgtgttctgg ctgccagaag cccatcaccg
     1501 gccgctgcat caccgccatg gccaagaagt tccaccccga gcacttcgtc tgtgccttct
     1561 gcctcaagca gctcaacaag ggcaccttca aggagcagaa cgacaagcct tactgtcaga
     1621 actgcttcct caagctcttc tgctagaatt catcgtgact gactgacgat ctgcctcgcg
     1681 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1741 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1801 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga
     1861 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1921 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1981 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2041 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2101 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2161 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2221 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2281 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     2341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2461 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2521 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2581 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     2641 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2701 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2761 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2821 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2881 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2941 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3841 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcataaa ttccgacacc atcgaatggc gcaaaacctt tcgcggtatg
     3961 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     4021 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     4081 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     4141 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     4201 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     4261 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     4321 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     4381 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     4441 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     4501 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     4561 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     4621 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     4681 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     4741 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     4801 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     4861 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     4921 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     4981 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5041 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5101 tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     5161 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5221 cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5281 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5341 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc
     5401 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg
     5461 tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga
     5521 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact
     5581 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg
     5641 atggcgttgg aatt
//
