LOCUS       sequence-192310-        6945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            DLG1-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6945)
  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 6945)
  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_DLG1-1/1
FEATURES             Location/Qualifiers
     source          1..6945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..47
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        70..147
                     /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             148..1230
                     /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(167..186)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1473..1478,1479..1494,1495..1501)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1509..1525
                     /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     1514..1536
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1610..1627
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1643..2296
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2254..2276
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2303..2326
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2339..2356
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2612..2659
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4252..4707)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4389..4410)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4601..4620
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4749..4771)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4857..4997
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4912..4931
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5012..5029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5183..5770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5941..6732,6733..6801))
                     /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     6564..6583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6802..6906)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
       61 agaattcccg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
      121 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
      181 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
      241 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
      301 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
      361 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
      421 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
      481 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
      541 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
      601 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
      661 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
      721 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
      781 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
      841 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
      901 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
      961 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     1021 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     1081 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     1141 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1201 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     1261 tcattaggca caattctcat gtttgacagc ttatcatcga ctgcacggtg caccaatgct
     1321 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca
     1381 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat
     1441 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat
     1501 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agccagtccg tttaggtgtt
     1561 ttcacgagca cttcaccaac aaggaccata gattatgaaa atcgaagaac accatcacca
     1621 tcaccattcc agcggtaagc ttatgtcccc tatactaggt tattggaaaa ttaagggcct
     1681 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
     1741 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
     1801 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
     1861 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
     1921 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
     1981 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
     2041 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
     2101 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
     2161 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
     2221 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
     2281 cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggcccctgt ccagcggtct
     2341 ggttccgcgt ggttccggta ccgcggccca gccggccatg gcccagggcg cgctgctgta
     2401 tgttcgtgcc ctgttcgatt atgacaaaac caaagattcc ggtctgccat ctcaaggtct
     2461 gaacttcaaa ttcggtgata ttctgcatgt cattaacgcc tccgatgatg aatggtggca
     2521 agcacgtcaa gttaccccag atggtgaatc cgatgaagtt ggtgttattc cgtccaaacg
     2581 tcgtgttgaa aaaaaagcgg ccgcataata aaccgataca attaaaggct ccttttggag
     2641 cctttttttt ggagattttc aacgtgaaaa aattattatt cgcaattcct ttagttgttc
     2701 ctttctattc tcactccgct gaaactgttg aaagttgttt agcaaaaccc catacagaaa
     2761 attcatttac taacgtctgg aaagacgaca aaactttaga tcgttacgct aactatgagg
     2821 gctgtctgtg gaatgctaca ggcgttgtag tttgtactgg tgacgaaact cagtgtctag
     2881 ctagagtggc ggtggctctg gttccggtga ttttgattat gaaaagatgg caaacgctaa
     2941 taagggggct atgaccgaaa atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa
     3001 acttgattct gtcgctactg attacggtgc tgctatcgat ggtttcattg gtgacgtttc
     3061 cggccttgct aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc
     3121 tcaagtcggt gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc
     3181 cctccctcaa tcggttgaat gtcgcccttt tgtctttggc gctggtaaac catatgaatt
     3241 ttctattgat tgtgacaaaa taaacttatt ccgtggtgtc tttgcgtttc ttttatatgt
     3301 tgccaccttt atgtatgtat tttctacgtt tgctaacata ctgcgtaata aggagtctta
     3361 atcatgccag ttcttttggc tagcgccgcc ctataccttg tctgcctccc cgcgttgcgt
     3421 cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg
     3481 gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa
     3541 ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca
     3601 tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga
     3661 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     3721 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     3781 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     3841 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     3901 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     3961 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     4021 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     4081 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     4141 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     4201 tctgtgaatc gcttcacgac cacgctgatg agctttaccg caggatccgc gaaattgtaa
     4261 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4321 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4381 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4441 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4501 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4561 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4621 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4681 cgcttaatgc gccgctacag ggcgcgtcgc ggatcctgcc tcgcgcgttt cggtgatgac
     4741 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4801 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     4861 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     4921 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4981 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5041 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5101 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5161 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5221 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac cagggtttcc
     5281 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5341 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5401 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5461 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5521 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5581 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5641 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5701 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5941 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6001 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6061 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6121 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6181 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6241 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6301 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatattgt gcaaaaaagc
     6361 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6421 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6481 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6541 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     6601 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6661 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6721 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6781 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6841 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6901 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaac
//
