LOCUS       sequence-365835-        7616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing recombinant paxillin variants in bacteria. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  AUTHORS   Case LB, De Pasquale M, Henry L, Rosen MK
  TITLE     Synergistic phase separation of two pathways promotes integrin 
            clustering and nascent adhesion formation.
  JOURNAL   Elife. 2022 Jan 20;11. pii: 72588. doi: 10.7554/eLife.72588.
  PUBMED    35049497
REFERENCE   2  (bases 1 to 7616)
  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: "Elife."; 
            date: "2022-01-20"; volume: "11. pii: 72588. doi"; pages: " 
            10.7554/eLife.72588"
COMMENT     Sequence Label: Paxillin DN
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        833..910
                     /gene="lacI"
                     /label=lacI promoter
     CDS             911..1993
                     /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(930..949)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2042..2047,2048..2065,2066..2072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2080..2096
                     /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     2104..2120
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2104..2120
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2116..2643
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2136..2153)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2136..2152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2145..2167)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2745..2750,2751..2766,2767..2773)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2781..2797
                     /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             2820..3473
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3431..3453
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3480..3497
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3498..3518
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(6228..6250)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6395..6413)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6481..6585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6586..6654,6655..7446)
                     /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(6804..6823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat
      841 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
      901 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      961 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1021 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1081 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1141 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1201 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1261 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1321 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     1381 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     1441 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     1501 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     1561 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     1621 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     1681 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1741 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     1801 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1861 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     1921 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1981 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     2041 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     2101 acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt
     2161 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2221 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2281 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     2341 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     2401 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     2461 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     2521 caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg
     2581 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     2641 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     2701 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     2761 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca
     2821 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
     2881 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
     2941 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
     3001 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
     3061 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     3121 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     3181 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     3241 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     3301 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     3361 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     3421 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
     3481 tggttccgcg tggatctgaa aacctgtatt ttcagggcca tatggacgat ctggacgcgc
     3541 tgttggccga ccttgaatcc accactagtc atattagtaa acgtccagta tttcttagcg
     3601 aggaaactcc ctatagttat ccaaccggaa atcataccta ccaggaaatc gccgtcccac
     3661 caccagtacc gcctccgcca tcctcggagg cgctgaacgg aactatcctt gatcctttag
     3721 atcagtggca acccagcggt tcccgcttca ttcatcaaca gccacaatca tcaagtcccg
     3781 tatatggtag tagtgcaaaa acgagcagcg tgagcaatcc tcaagattct gtaggttcgc
     3841 cctgtagtcg cgtcggagaa gaggaacacg tttattcctt ccctaataag cagaaaagcg
     3901 cggaaccctc gccaacagtc atgtctacgt cgttgggctc gaacctgtct gagcttgacc
     3961 gcctgctgtt agagcttaac gccgtacaac ataaccctcc tggtttcccg gccgacgagg
     4021 ctaacagctc cccaccctta ccaggtgccc ttagcccatt gtacggggta cctgaaacaa
     4081 attcacctct tggcgggaaa gcagggcctc tgacgaaaga aaaaccgaaa cgcaatgggg
     4141 gtcgtgggtt ggaggatgta cgtccatcag tagagtccct tctggacgag ctggagagtt
     4201 ctgtcccatc cccggttccc gccattactg tcaatcaagg agaaatgtca tccccccaac
     4261 gcgtgaccag tactcaacaa cagacccgta tttccgcctc gagcgctaca cgtgaattag
     4321 atgaattaat ggcttcgctt tcggatttta aattcatggc tcagggaaag accggtagtt
     4381 cctcaccgcc cggtggaccg cctaaaccag gcagtcaatt agactctatg ctgggatcat
     4441 tgcagagcga tttaaataag ttgggggtcg cgacagtcgc aaagatggat gacttggatg
     4501 ctttacttgc agatttggaa tctaccactt cccacattag caagcgtccg gtatttttgt
     4561 ccgaggagac accttatagc taccccactg gtaaccacac ttatcaggag attgcggtac
     4621 caccccctgt gccaccacct ccttcttccg aggccctgaa tggcacgatc cttgatccct
     4681 tggatcaatg gcagccatcc gggtctcgtt ttatccacca acaaccgcag agttcgtccc
     4741 ccgtctatgg gtcctccgcg aagacgagct cagtctcgaa ccctcaagat tctgtcgggt
     4801 ctccatgtag ccgtgtgggt gaggaagaac atgtgtactc attccccaac aagcaaaaaa
     4861 gtgcggaacc atcgcctact gtcatgtcga caagtcttgg tagcaacctt tccgaacttg
     4921 atcgtttgtt acttgaactg aatgcggtcc aacataaccc gcctggattt ccggctgacg
     4981 aagctaactc ttcgccacct ctgccgggag cgctgtctcc cctgtatgga gtgcccgaga
     5041 ccaattcacc attgggtggt aaggcgggac cgcttaccaa agaaaagccg aagcgcaacg
     5101 ggggccgtgg attagaagac gttcgccctt ccgtggaatc cttacttgat gagctggaga
     5161 gctctgtgcc atcaccggtg cctgccatta cagtcaatca gggcgaaatg tccagccctc
     5221 aacgcgttac tagtacccaa cagcaaactc gcatttccgc ctcgagcgct acacgcgaat
     5281 tagatgagtt gatggccagc ctttccgatt tcaaattcat ggcgcaaggt aaaaccggta
     5341 gtagctcacc accaggtggg ccaccgaagc cgggctctca acttgactcg atgttaggta
     5401 gcttgcagag tgaccttaac aaattaggtg tcgcaacagt agcaaaaggg gtgtgcggag
     5461 cctgcaagaa gccgattgca ggccaagttg tcactgcgat gggtaagaca tggcaccctg
     5521 agcactttgt ctgcactcat tgtcaggagg agattggttc ccgtaatttc tttgagcgcg
     5581 atgggcagcc ctactgcgag aaagactacc ataacttatt ttccccccgc tgctactatt
     5641 gcaatggccc gatcttagat aaagtcgtca ccgcactgga tcgcacatgg caccccgaac
     5701 atttcttttg cgcccagtgt ggtgcttttt ttggacctga aggctttcac gaaaaggatg
     5761 ggaaagcata ctgtcgtaaa gattactttg acatgtttgc accgaaatgt ggggggtgtg
     5821 cacgcgcgat cctggagaac tacattagcg cgctgaatac gttatggcat cctgagtgtt
     5881 ttgtctgtcg tgagtgtttt acacccttcg tcaatggttc attcttcgaa cacgacggcc
     5941 agccgtattg tgaagtccac taccatgaac gccgcggttc gttatgttca ggttgccaga
     6001 aacctattac cggtcgctgt attactgcca tggccaagaa gtttcatcca gagcactttg
     6061 tgtgcgcatt ttgtttgaaa cagctgaata aagggacatt taaggaacag aatgataagc
     6121 cctactgcca aaattgtttt cttaaactgt tttgctagtg aggatccgcg gccgcatcgt
     6181 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6241 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6301 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     6361 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     6421 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     6841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     7141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
