LOCUS       sequence-192532-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            DLG3-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  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 6960)
  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_DLG3-1/1
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..39
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        62..139
                     /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             140..1222
                     /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(159..178)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1465..1470,1471..1486,1487..1493)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1501..1517
                     /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     1506..1528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1602..1619
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1635..2288
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2246..2268
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2295..2318
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2331..2348
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2619..2666
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4259..4714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4608..4627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4756..4778)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4864..5004
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4919..4938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5019..5036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5190..5777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5948..6739,6740..6808))
                     /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     6571..6590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6809..6913)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattcc
       61 cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
      121 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      181 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      241 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
      301 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
      361 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
      421 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
      481 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
      541 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
      601 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
      661 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
      721 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
      781 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
      841 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
      901 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
      961 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     1021 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     1081 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     1141 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1201 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg
     1261 cacaattctc atgtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt
     1321 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt
     1381 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     1441 tggcaaatat tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa
     1501 ttgtgagcgg ataacaattt cacacaggaa acagccagtc cgtttaggtg ttttcacgag
     1561 cacttcacca acaaggacca tagattatga aaatcgaaga acaccatcac catcaccatt
     1621 ccagcggtaa gcttatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
     1681 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
     1741 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
     1801 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
     1861 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
     1921 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
     1981 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
     2041 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
     2101 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
     2161 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
     2221 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
     2281 ctccaaaatc ggatctggaa gttctgttcc aggggcccct gtccagcggt ctggttccgc
     2341 gtggttccgg taccgcggcc cagccggcct tatatgttcg tgccctgttc gattatgatc
     2401 gtacccgtga ttcttgtctg ccatcccagg gtctgagttt ttcctatggt gatattctgc
     2461 atgtcattaa cgcctccgat gatgaatggt ggcaagcacg tttagttacc ccacatggtg
     2521 aatccgaaca gattggtgtt attccgtcca aaaaacgtgt cgaaaaaaaa gcggccgctt
     2581 cacgtttctg gacatagttg ctagcggccg cataataaac cgatacaatt aaaggctcct
     2641 tttggagcct tttttttgga gattttcaac gtgaaaaaat tattattcgc aattccttta
     2701 gttgttcctt tctattctca ctccgctgaa actgttgaaa gttgtttagc aaaaccccat
     2761 acagaaaatt catttactaa cgtctggaaa gacgacaaaa ctttagatcg ttacgctaac
     2821 tatgagggct gtctgtggaa tgctacaggc gttgtagttt gtactggtga cgaaactcag
     2881 tgtctagcta gagtggcggt ggctctggtt ccggtgattt tgattatgaa aagatggcaa
     2941 acgctaataa gggggctatg accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta
     3001 aaggcaaact tgattctgtc gctactgatt acggtgctgc tatcgatggt ttcattggtg
     3061 acgtttccgg ccttgctaat ggtaatggtg ctactggtga ttttgctggc tctaattccc
     3121 aaatggctca agtcggtgac ggtgataatt cacctttaat gaataatttc cgtcaatatt
     3181 taccttccct ccctcaatcg gttgaatgtc gcccttttgt ctttggcgct ggtaaaccat
     3241 atgaattttc tattgattgt gacaaaataa acttattccg tggtgtcttt gcgtttcttt
     3301 tatatgttgc cacctttatg tatgtatttt ctacgtttgc taacatactg cgtaataagg
     3361 agtcttaatc atgccagttc ttttggctag cgccgcccta taccttgtct gcctccccgc
     3421 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     3481 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     3541 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     3601 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     3661 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     3721 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     3781 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     3841 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     3901 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     3961 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     4021 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     4081 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     4141 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4201 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag gatccgcgaa
     4261 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4321 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4381 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4441 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4501 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4561 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4621 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4681 cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgga tcctgcctcg cgcgtttcgg
     4741 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4801 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4861 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     4921 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4981 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5041 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5101 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5281 ggtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6241 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6541 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     6601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
//
