LOCUS       sequence-191132-        6927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            GAS7-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  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 6927)
  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_GAS7-1/1
FEATURES             Location/Qualifiers
     source          1..6927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             34..687
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     645..667
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             694..717
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             730..747
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      985..1032
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(2625..3080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2762..2783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2974..2993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3122..3144)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3230..3370
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3285..3304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3385..3402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3556..4143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4314..5105,5106..5174))
                     /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     4937..4956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5175..5279)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5347..5365
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5388..5465
                     /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             5466..6548
                     /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(5485..5504)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6791..6796,6797..6812,6813..6819)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6827..6843
                     /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     6832..6854
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caccatcacc atcaccattc cagcggtaag cttatgtccc ctatactagg ttattggaaa
       61 attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa
      121 gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt
      181 ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg
      241 gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt
      301 gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt
      361 gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg
      421 ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc
      481 catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc
      541 ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt
      601 gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg
      661 tttggtggtg gcgaccatcc tccaaaatcg gatctggaag ttctgttcca ggggcccctg
      721 tccagcggtc tggttccgcg tggttccggt accgcggccc agccggccat gagtggtgcc
      781 cgttgtcgta cgctgtatcc gttttcgggt gaacgtcatg gtcaaggtct gcgctttgct
      841 gctggtgaac tgattaccct gctgcaggtc ccggatggcg gttggtggga aggcgaaaaa
      901 gaagacggcc tgcgtggttg gtttccggcg agctatgtgc aactgctgga aaaaccgggt
      961 atggttccgg cggccgcata ataaaccgat acaattaaag gctccttttg gagccttttt
     1021 tttggagatt ttcaacgtga aaaaattatt attcgcaatt cctttagttg ttcctttcta
     1081 ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa ccccatacag aaaattcatt
     1141 tactaacgtc tggaaagacg acaaaacttt agatcgttac gctaactatg agggctgtct
     1201 gtggaatgct acaggcgttg tagtttgtac tggtgacgaa actcagtgtc tagctagagt
     1261 ggcggtggct ctggttccgg tgattttgat tatgaaaaga tggcaaacgc taataagggg
     1321 gctatgaccg aaaatgccga tgaaaacgcg ctacagtctg acgctaaagg caaacttgat
     1381 tctgtcgcta ctgattacgg tgctgctatc gatggtttca ttggtgacgt ttccggcctt
     1441 gctaatggta atggtgctac tggtgatttt gctggctcta attcccaaat ggctcaagtc
     1501 ggtgacggtg ataattcacc tttaatgaat aatttccgtc aatatttacc ttccctccct
     1561 caatcggttg aatgtcgccc ttttgtcttt ggcgctggta aaccatatga attttctatt
     1621 gattgtgaca aaataaactt attccgtggt gtctttgcgt ttcttttata tgttgccacc
     1681 tttatgtatg tattttctac gtttgctaac atactgcgta ataaggagtc ttaatcatgc
     1741 cagttctttt ggctagcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
     1801 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
     1861 cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc
     1921 ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg
     1981 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     2041 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     2101 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     2161 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     2221 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     2281 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     2341 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     2401 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     2461 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     2521 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     2581 atcgcttcac gaccacgctg atgagcttta ccgcaggatc cgcgaaattg taagcgttaa
     2641 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2701 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2761 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2821 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2881 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2941 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3001 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3061 tgcgccgcta cagggcgcgt cgcggatcct gcctcgcgcg tttcggtgat gacggtgaaa
     3121 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3181 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     3241 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     3301 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3361 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3421 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3481 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccagggtt tccccctgga
     3661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3721 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3961 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4321 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4381 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4441 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4501 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4561 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4621 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4681 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4741 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4801 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4861 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4921 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4981 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5041 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5101 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5161 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5221 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5281 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5341 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattcccgac accatcgaat
     5401 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     5461 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     5521 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     5581 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     5641 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     5701 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     5761 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     5821 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     5881 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     5941 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     6001 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     6061 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     6121 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     6181 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     6241 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     6301 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     6361 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     6421 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     6481 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6541 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcaca attctcatgt
     6601 ttgacagctt atcatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg
     6661 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
     6721 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
     6781 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
     6841 caatttcaca caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa
     6901 ggaccataga ttatgaaaat cgaagaa
//
