LOCUS       sequence-236082-        7889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid containing His and MBP tags for 5 
            repeats of the second SH3 domain from human NCK1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  AUTHORS   Li P, Banjade S, Cheng HC, Kim S, Chen B, Guo L, Llaguno M, 
            Hollingsworth JV, King DS, Banani SF, Russo PS, Jiang QX, Nixon BT, 
            Rosen MK
  TITLE     Phase transitions in the assembly of multivalent signalling 
            proteins.
  JOURNAL   Nature. 2012 Mar 7;483(7389):336-40. doi: 10.1038/nature10879.
  PUBMED    22398450
REFERENCE   2  (bases 1 to 7889)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2012-03-7- 7"; volume: "483"; issue: "7389"; pages:
            "336-40"
COMMENT     Sequence Label: pMAL-SH3(2)-5R
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             123..1223
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1080..1089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1197..1220
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1272..1283
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1296..1316
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2526..2546
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2547..2564
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2577..2594)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2577..2593)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2586..2608)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2772..2789)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2942..3028
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3120..3147
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3166..3257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3258..3326,3327..4118)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3476..3495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4160..4673)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4355..4376)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4567..4586
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4784..5372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5273..5292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5526..5543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5558..5698
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5624..5643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5784..5806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5800..5991)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6487..6564
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6565..7647
                     /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(6584..6603)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaac tgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg
      181 gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc
      241 atccggataa actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca
      301 ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa
      361 tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt
      421 acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca
      481 aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac
      541 tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc
      601 cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta
      661 aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga
      721 ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata
      781 aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca
      841 aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg
      901 ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt
      961 tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc
     1021 tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg
     1081 ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt
     1141 tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg
     1201 aagccctgaa agacgcgcag actaattcga gctcgaacaa caacaacaat aacaataaca
     1261 acaacctcgg gatcgaggga aggatttcag aatttgaaaa cctgtatttt cagggccata
     1321 tggacctcaa catgcccgct tatgtgaaat ttaactacat ggctgagaga gaggatgaat
     1381 tatcattgat aaaggggaca aaggtgatcg tcatggagaa aagcagtgat gggtggtggc
     1441 gtggtagcta caatggacaa gttggatggt tcccttcaaa ctatgtaact gaagaaggtg
     1501 acagtccttt ggcgtctggt gcgggtggtt ctgaaggtgg tggttctgaa ggtggtacct
     1561 ctggtgcgac cgacctcaac atgcccgctt atgtgaaatt taactacatg gctgagagag
     1621 aggatgaatt atcattgata aaggggacaa aggtgatcgt catggagaaa agcagtgatg
     1681 ggtggtggcg tggtagctac aatggacaag ttggatggtt cccttcaaac tatgtaactg
     1741 aagaaggtga cagtcctttg gcatccggag caggaggctc cgaaggagga ggctccgagg
     1801 gaggaacatc cggagcaaca catatggacc tcaacatgcc cgcttatgtg aaatttaact
     1861 acatggctga gagagaggat gaattatcat tgataaaggg gacaaaggtg atcgtcatgg
     1921 agaaaagcag tgatgggtgg tggcgtggta gctacaatgg acaagttgga tggttccctt
     1981 caaactatgt aactgaagaa ggtgacagtc ctttggcgtc tggtgcgggt ggttctgaag
     2041 gtggtggttc tgaaggtggt acctctggtg cgaccgacct caacatgccc gcttatgtga
     2101 aatttaacta catggctgag agagaggatg aattatcatt gataaagggg acaaaggtga
     2161 tcgtcatgga gaaaagcagt gatgggtggt ggcgtggtag ctacaatgga caagttggat
     2221 ggttcccttc aaactatgta actgaagaag gtgacagtcc tttggcatcc ggagcaggag
     2281 gctccgaagg aggaggctcc gagggaggaa catccggagc aacagacctc aacatgcccg
     2341 cttatgtgaa atttaactac atggctgaga gagaggatga attatcattg ataaagggga
     2401 caaaggtgat cgtcatggag aaaagcagtg atgggtggtg gcgtggtagc tacaatggac
     2461 aagttggatg gttcccttca aactatgtaa ctgaagaagg tgacagtcct ttgggaggcg
     2521 gatccgaaaa cctgtatttt cagggccatc atcatcatca tcactgaaag cttggcactg
     2581 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2641 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2701 tcccaacagt tgcgcagcct gaatggcgaa tggcagcttg gctgttttgg cggatgagat
     2761 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     2821 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     2881 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2941 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     3001 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     3061 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     3121 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     3181 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3241 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3301 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3361 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3421 ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa gttctgctat
     3481 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     3541 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3601 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3661 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3721 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3781 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3841 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3901 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3961 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4021 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4081 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4141 atatacttta gattgattta ccccggttga taatcagaaa agccccaaaa acaggaagat
     4201 tgtataagca aatatttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     4261 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4321 aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4381 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4441 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4501 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4561 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4621 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga
     4681 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4741 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4801 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4861 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4921 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4981 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5041 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5101 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5161 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5221 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5281 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5341 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5401 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5461 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5521 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     5581 cgcatctgtg cggtatttca caccgcatgt ggtgcactct cagtacaatc tgctctgatg
     5641 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     5701 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5761 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5821 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca
     5881 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5941 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     6001 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     6061 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     6121 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     6181 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     6241 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     6301 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     6361 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     6421 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     6481 aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     6541 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     6601 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     6661 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     6721 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     6781 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     6841 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     6901 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     6961 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     7021 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     7081 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     7141 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     7201 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     7261 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     7321 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     7381 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     7441 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     7501 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     7561 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7621 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     7681 ttaggcacaa ttctcatgtt tgacagctta tcatcgactg cacggtgcac caatgcttct
     7741 ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa
     7801 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     7861 ggttctggca aatattctga aatgagctg
//
