LOCUS       sequence-191059-        6927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            DLG4-1/1.
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_DLG4-1/1
FEATURES             Location/Qualifiers
     source          1..6927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(159..227,228..1019)
                     /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     complement(377..396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1190..1777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1931..1948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1963..2103
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2029..2048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2189..2211
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2253..2708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2340..2359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2550..2571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(4301..4348)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             4559..4570
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(4586..4603)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4616..4639)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(4646..5299)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(4666..4688)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(5315..5332)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5406..5428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5417..5433
                     /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)."
     promoter        complement(join(5441..5447,5448..5463,5464..5469))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(5712..6794)
                     /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     6756..6775
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6795..6872)
                     /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."
     primer_bind     complement(6895..6913)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
       61 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      121 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      181 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      241 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      301 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
      361 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
      421 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
      481 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
      541 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
      601 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
      661 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
      721 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
      781 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
      841 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
      901 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
      961 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1021 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1081 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1141 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1201 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1261 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1321 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1381 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1441 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1501 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1561 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1621 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1681 gaaaccctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1741 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1801 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1861 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     1921 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     1981 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     2041 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     2101 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2161 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2221 tcatcaccga aacgcgcgag gcaggatccg cgacgcgccc tgtagcggcg cattaagcgc
     2281 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2341 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2401 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2461 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2521 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2581 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2641 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     2701 tacaatttcg cggatcctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     2761 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     2821 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     2881 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     2941 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     3001 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3061 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     3121 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     3181 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     3241 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     3301 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     3361 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     3421 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     3481 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     3541 cgcaacgcgg ggaggcagac aaggtatagg gcggcgctag ccaaaagaac tggcatgatt
     3601 aagactcctt attacgcagt atgttagcaa acgtagaaaa tacatacata aaggtggcaa
     3661 catataaaag aaacgcaaag acaccacgga ataagtttat tttgtcacaa tcaatagaaa
     3721 attcatatgg tttaccagcg ccaaagacaa aagggcgaca ttcaaccgat tgagggaggg
     3781 aaggtaaata ttgacggaaa ttattcatta aaggtgaatt atcaccgtca ccgacttgag
     3841 ccatttggga attagagcca gcaaaatcac cagtagcacc attaccatta gcaaggccgg
     3901 aaacgtcacc aatgaaacca tcgatagcag caccgtaatc agtagcgaca gaatcaagtt
     3961 tgcctttagc gtcagactgt agcgcgtttt catcggcatt ttcggtcata gcccccttat
     4021 tagcgtttgc catcttttca taatcaaaat caccggaacc agagccaccg ccactctagc
     4081 tagacactga gtttcgtcac cagtacaaac tacaacgcct gtagcattcc acagacagcc
     4141 ctcatagtta gcgtaacgat ctaaagtttt gtcgtctttc cagacgttag taaatgaatt
     4201 ttctgtatgg ggttttgcta aacaactttc aacagtttca gcggagtgag aatagaaagg
     4261 aacaactaaa ggaattgcga ataataattt tttcacgttg aaaatctcca aaaaaaaggc
     4321 tccaaaagga gcctttaatt gtatcggttt attatgcggc cgcgcgacgt tcgacgcgac
     4381 gtttgctcgg gataaagcca atgtcatcgg tttcagaatc gctgtgaacg cgacgggcct
     4441 gccaccattc ttcgtcactc gcatcaatca catgcagcac atcgccgaaa cggaatgaca
     4501 gggcttggct caggaaacca cagtctttgg ttttgtcgta gtcgaacaga gcacggatat
     4561 agaaggccgg ctgggccgcg gtaccggaac cacgcggaac cagaccgctg gacaggggcc
     4621 cctggaacag aacttccaga tccgattttg gaggatggtc gccaccacca aacgtggctt
     4681 gccagccctg caaaggccat gctatatact tgctggattt caagtactta tcaatttgtg
     4741 ggatagcttc aatacgtttt ttaaaacaaa ctaattttgg gaacgcatcc aggcacattg
     4801 ggtccatgta taaaacaaca tcaagagcgt catacaacat gaagtcagga tgggttacat
     4861 gatcaccatt taaatatgtt ttatgacata aacgatcttc gaacattttc agcatttcag
     4921 gtagcttgct aagaaaatca actttgagag tttcaaagtc tttactatat gcaattctcg
     4981 aaacaccgta tctaatatcc aaaaccgctc cttcaagcat tgaaatctct gcacgctctt
     5041 ttggacaacc acccaacatg ttgtgcttgt cagctatata acgtatgatg gccatagact
     5101 gtgttaattt aacatcacca tcaatataat aaggaagatt gggaaactcc aaacccaatt
     5161 caaacttttt gtttcgccat ttatcacctt catcgcgctc atacaaatgc tcttcatatt
     5221 tttcttcaag atattccaaa agaagtcgag tgggttgcac aaggccctta attttccaat
     5281 aacctagtat aggggacata agcttaccgc tggaatggtg atggtgatgg tgttcttcga
     5341 ttttcataat ctatggtcct tgttggtgaa gtgctcgtga aaacacctaa acggactggc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca cattatacga gccgatgatt
     5461 aattgtcaac agctcatttc agaatatttg ccagaaccgt tatgatgtcg gcgcaaaaaa
     5521 cattatccag aacgggagtg cgccttgagc gacacgaatt atgcagtgat ttacgacctg
     5581 cacagccata ccacagcttc cgatggctgc ctgacgccag aagcattggt gcaccgtgca
     5641 gtcgatgata agctgtcaaa catgagaatt gtgcctaatg agtgagctaa cttacattaa
     5701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5761 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     5821 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     5881 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     5941 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     6001 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     6061 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     6121 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     6181 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     6241 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     6301 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     6361 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     6421 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     6481 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     6541 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     6601 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     6661 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     6721 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     6781 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     6841 ccgcgaaagg ttttgcacca ttcgatggtg tcgggaattc ttgaagacga aagggcctcg
     6901 tgatacgcct atttttatag gttaatg
//
