LOCUS       sequence-358841-        5654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of human paxillin LIM domains 1, 2, 2 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_LIM1224
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..132
                     /gene="lacI"
                     /label=lacI promoter
     CDS             133..1215
                     /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(152..171)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1264..1269,1270..1287,1288..1294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1302..1318
                     /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     1326..1342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1326..1342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1338..1865
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1358..1375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1358..1374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1367..1389)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1967..1972,1973..1988,1989..1995)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2003..2019
                     /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             2042..2695
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2653..2675
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2702..2719
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3488..3510)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3655..3673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3741..3845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3846..3914,3915..4706)
                     /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(4064..4083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4877..5465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5366..5385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5619..5636
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
       61 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
      121 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
      181 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
      241 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
      301 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
      361 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
      421 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
      481 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
      541 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
      601 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
      661 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
      721 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
      781 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
      841 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
      901 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
      961 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     1021 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     1081 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     1141 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     1201 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     1261 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     1321 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     1381 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1441 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1501 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     1561 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     1621 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
     1681 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     1741 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
     1801 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
     1861 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
     1921 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca
     1981 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt
     2041 catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
     2101 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
     2161 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
     2221 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
     2281 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
     2341 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
     2401 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
     2461 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
     2521 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
     2581 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
     2641 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga
     2701 tctggttccg cgtggatctg gagtctgcgg ggcctgcaag aagcccatcg ccgggcaggt
     2761 tgtgaccgcc atggggaaga cgtggcaccc cgagcacttc gtctgcaccc actgccagga
     2821 ggagatcgga tcccggaact tcttcgagcg ggatggacag ccctactgtg aaaaggacta
     2881 ccacaacctc ttctcccctc gctgctatta ctgtaacgga cccatcctcg ataaagtggt
     2941 cacagccctt gaccggacgt ggcaccctga acacttcttt tgcgcacaat gtggcgcctt
     3001 ctttggtcct gagggatttc atgagaagga cggcaaagcc tattgtcgga aggattattt
     3061 cgacatgttt gcaccccggt gttactattg caatggccct attctggaca aggtcgtgac
     3121 ggcactcgat cgcacatggc atccagagca ttttttctgt gctcagtgcg gagctttttt
     3181 cggacccgaa gggttccacg aaaaagatgg aaaggcttac tgccgcaagg actactttga
     3241 tatgttcggc tcgctgtgtt ctggctgcca gaagcccatc accggccgct gcatcaccgc
     3301 catggccaag aagttccacc ccgagcactt cgtctgtgcc ttctgcctca agcagctcaa
     3361 caagggcacc ttcaaggagc agaacgacaa gccttactgt cagaactgct tcctcaagct
     3421 cttctgctag aattcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg
     3481 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     3541 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     3601 ccatgaccca gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg
     3661 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3721 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3781 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3841 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3901 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3961 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4021 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4081 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4141 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4201 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4261 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4321 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4381 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4441 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4501 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4561 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4621 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4681 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4801 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5521 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5581 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5641 cggaagagcg cctg
//
