LOCUS       sequence-365817-        6110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of recombinant Nck in bacteria cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6110)
  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 6110)
  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: Nck
FEATURES             Location/Qualifiers
     source          1..6110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(583..588,589..606,607..613)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    621..637
                     /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     645..661
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     645..661
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             657..1184
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(677..694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(677..693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(686..708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1286..1291,1292..1307,1308..1314)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1322..1338
                     /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             1361..2014
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1972..1994
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2021..2038
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2039..2059
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3263..3285)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3430..3448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3516..3620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3621..3689,3690..4481)
                     /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(3839..3858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4652..5240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5141..5160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5394..5411
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5484..5561
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5562..6110,1..534)
                     /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(5581..5600)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
       61 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
      121 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      181 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      241 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
      301 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
      361 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
      421 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      481 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      541 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      601 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      661 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
      841 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
      901 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
      961 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     1021 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     1081 ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg
     1141 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1201 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1261 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1321 attgtgagcg gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta
     1381 ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa
     1441 atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga
     1501 attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca
     1561 gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa
     1621 agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc
     1681 gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc
     1741 tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca
     1801 tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc
     1861 aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc
     1921 acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca
     1981 agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggttccgc gtggatctga
     2041 aaacctgtat tttcagggcc atatggcaga agaagtggtg gtagtagcca aatttgatta
     2101 tgtggcccaa caagaacaag agttggacat caagaagaat gagagattat ggcttctgga
     2161 tgattctaag tcctggtggc gagttcgaaa ttccatgaat aaaacaggtt ttgtgccttc
     2221 taactatgtg gaaaggaaaa acagtgctcg gaaagcatct attgtgaaaa acctaaagga
     2281 taccttaggc attggaaaag tgaaaagaaa acctagtgtg ccagattctg catctcctgc
     2341 tgatgatagt tttgttgacc caggggaacg tctctatgac ctcaacatgc ccgcttatgt
     2401 gaaatttaac tacatggctg agagagagga tgaattatca ttgataaagg ggacaaaggt
     2461 gatcgtcatg gagaaaagca gtgatgggtg gtggcgtggt agctacaatg gacaagttgg
     2521 atggttccct tcaaactatg taactgaaga aggtgacagt cctttgggtg accatgtggg
     2581 ttctctgtca gagaaattag cagcagtcgt caataaccta aatactgggc aagtgttgca
     2641 tgtggtacag gctctttacc cattcagctc atctaatgat gaagaactta atttcgagaa
     2701 aggagatgta atggatgtta ttgaaaaacc tgaaaatgac ccagagtggt ggaaagcgag
     2761 gaagatcaat ggtatggttg gtctagtacc aaaaaactat gttaccgtta tgcagaataa
     2821 tccattaact tcaggtttgg aaccatcacc tccacagagc gattacatta ggccttcact
     2881 cactggaaag tttgctggca atccttggta ttatggcaaa gtcaccaggc atcaagcaga
     2941 aatggcatta aatgaaagag gacatgaagg ggatttcctc attcgtgata gtgaatcttc
     3001 gccaaatgat ttctcagtat cactaaaagc acaagggaaa aacaagcatt ttaaagtcca
     3061 actaaaagag actgtctaca gtattgggca gcgtaaattc agcaccatgg aagaacttgt
     3121 agaacattac aaaaaggcac caatttttac aagtgaacaa ggagaaaaat tatatcttgt
     3181 caagcattta tcatgaggat ccgcggccgc atcgtgactg actgacgatc tgcctcgcgc
     3241 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3301 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3361 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa
     3421 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3481 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3541 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3601 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3661 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3721 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3781 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3841 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3901 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3961 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4021 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4081 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4141 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     4201 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4261 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4321 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4381 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4441 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4501 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4561 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4621 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4681 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4741 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4801 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4861 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4981 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5041 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5101 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5161 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5221 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5281 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5341 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5401 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5461 gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg
     5521 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     5581 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     5641 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     5701 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     5761 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     5821 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     5881 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     5941 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     6001 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     6061 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
//
