LOCUS       sequence-192536-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            STAC-1/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  AUTHORS   Teyra J, Huang H, Jain S, Guan X, Dong A, Liu Y, Tempel W, Min J, 
            Tong Y, Kim PM, Bader GD, Sidhu SS
  TITLE     Comprehensive Analysis of the Human SH3 Domain Family Reveals a Wide
            Variety of Non-canonical Specificities.
  JOURNAL   Structure. 2017 Oct 3;25(10):1598-1610.e3. doi: 
            10.1016/j.str.2017.07.017. Epub 2017 Sep 7.
  PUBMED    28890361
REFERENCE   2  (bases 1 to 6894)
  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: 
            "10.1016/j.str.2017.07.017"; journalName: "Structure"; date: 
            "2017-10-3- 3"; volume: "25"; issue: "10"; pages: "1598-1610.e3"
COMMENT     Sequence Label: pHH0103_STAC-1/2
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1169..1174,1175..1190,1191..1197)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1205..1221
                     /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     1210..1232
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1306..1323
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1339..1992
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1950..1972
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1999..2022
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2035..2052
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2257..2304
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3897..4352)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4034..4055)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4246..4265
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4394..4416)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4502..4642
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4557..4576
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4657..4674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4828..5415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5586..6377,6378..6446))
                     /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     6209..6228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6447..6551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6619..6637
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6660..6737
                     /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             join(6738..6894,1..926)
                     /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(6757..6776)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
       61 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
      121 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
      181 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
      241 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
      301 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
      361 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
      421 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
      481 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
      541 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
      601 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
      661 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
      721 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
      781 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
      841 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      901 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
      961 taggcacaat tctcatgttt gacagcttat catcgactgc acggtgcacc aatgcttctg
     1021 gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat
     1081 tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg
     1141 gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt
     1201 ggaattgtga gcggataaca atttcacaca ggaaacagcc agtccgttta ggtgttttca
     1261 cgagcacttc accaacaagg accatagatt atgaaaatcg aagaacacca tcaccatcac
     1321 cattccagcg gtaagcttat gtcccctata ctaggttatt ggaaaattaa gggccttgtg
     1381 caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag
     1441 cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat
     1501 cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat
     1561 atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg
     1621 cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac
     1681 tttgaaactc tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa
     1741 gatcgtttat gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg
     1801 ttgtatgacg ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca
     1861 aaattagttt gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa
     1921 tccagcaagt atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac
     1981 catcctccaa aatcggatct ggaagttctg ttccaggggc ccctgtccag cggtctggtt
     2041 ccgcgtggtt ccggtaccgc ggcccagccg gccaacacct atgtcgccct gtacaaattc
     2101 gttccgcagg aaaacgaaga tttggaaatg cgtccgggtg acattatcac cctgctggaa
     2161 gattccaatg aagattggtg gaaaggtaaa attcaggatc gtattggttt ctttccggcc
     2221 aactttgttc aacgtctgca agcggccgca taataaaccg atacaattaa aggctccttt
     2281 tggagccttt tttttggaga ttttcaacgt gaaaaaatta ttattcgcaa ttcctttagt
     2341 tgttcctttc tattctcact ccgctgaaac tgttgaaagt tgtttagcaa aaccccatac
     2401 agaaaattca tttactaacg tctggaaaga cgacaaaact ttagatcgtt acgctaacta
     2461 tgagggctgt ctgtggaatg ctacaggcgt tgtagtttgt actggtgacg aaactcagtg
     2521 tctagctaga gtggcggtgg ctctggttcc ggtgattttg attatgaaaa gatggcaaac
     2581 gctaataagg gggctatgac cgaaaatgcc gatgaaaacg cgctacagtc tgacgctaaa
     2641 ggcaaacttg attctgtcgc tactgattac ggtgctgcta tcgatggttt cattggtgac
     2701 gtttccggcc ttgctaatgg taatggtgct actggtgatt ttgctggctc taattcccaa
     2761 atggctcaag tcggtgacgg tgataattca cctttaatga ataatttccg tcaatattta
     2821 ccttccctcc ctcaatcggt tgaatgtcgc ccttttgtct ttggcgctgg taaaccatat
     2881 gaattttcta ttgattgtga caaaataaac ttattccgtg gtgtctttgc gtttctttta
     2941 tatgttgcca cctttatgta tgtattttct acgtttgcta acatactgcg taataaggag
     3001 tcttaatcat gccagttctt ttggctagcg ccgccctata ccttgtctgc ctccccgcgt
     3061 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
     3121 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     3181 gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg
     3241 gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt
     3301 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     3361 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     3421 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     3481 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     3541 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     3601 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     3661 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     3721 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     3781 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     3841 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagga tccgcgaaat
     3901 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3961 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4021 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4081 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4141 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4201 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4261 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4321 cgccgcgctt aatgcgccgc tacagggcgc gtcgcggatc ctgcctcgcg cgtttcggtg
     4381 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4441 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4501 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     4561 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4621 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggg
     4921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5701 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5881 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6181 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     6241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6421 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6481 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6541 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6601 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattcccg
     6661 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     6721 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     6781 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     6841 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtgg
//
