LOCUS       sequence-192623-        6933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            MPP5-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6933)
  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 6933)
  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_MPP5-1/1
FEATURES             Location/Qualifiers
     source          1..6933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..112
                     /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             113..1195
                     /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(132..151)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1438..1443,1444..1459,1460..1466)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1474..1490
                     /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     1479..1501
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1575..1592
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1608..2261
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2219..2241
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2268..2291
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2304..2321
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2565..2612
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4205..4660)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4342..4363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4554..4573
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4702..4724)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4810..4950
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4865..4884
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4965..4982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5136..5723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5894..6685,6686..6754))
                     /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     6517..6536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6755..6859)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(6927..6933,1..12)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtatcacgag gccctttcgt cttcaagaat tcccgacacc atcgaatggt gcaaaacctt
       61 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
      121 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
      181 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
      241 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
      301 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
      361 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
      421 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
      481 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
      541 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
      601 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
      661 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
      721 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
      781 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
      841 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
      901 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
      961 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     1021 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     1081 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1141 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1201 acgcaattaa tgtaagttag ctcactcatt aggcacaatt ctcatgtttg acagcttatc
     1261 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     1321 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     1381 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     1441 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     1501 gaaacagcca gtccgtttag gtgttttcac gagcacttca ccaacaagga ccatagatta
     1561 tgaaaatcga agaacaccat caccatcacc attccagcgg taagcttatg tcccctatac
     1621 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
     1681 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     1741 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     1801 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     1861 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     1921 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     1981 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     2041 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     2101 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     2161 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     2221 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gaagttctgt
     2281 tccaggggcc cctgtccagc ggtctggttc cgcgtggttc cggtaccgcg gcccagccgg
     2341 ccattcacgt caaagcccat ttcgattacg atccatccga tgatccatac gttccatgtc
     2401 gtgaactggg tctgagcttc cagaaaggtg atattctgca tgtcatttcc caggaagatc
     2461 caaattggtg gcaagcctac cgtgaaggtg atgaagataa tcaaccactg gcaggtctgg
     2521 ttcctggtaa atcctttcaa cagcagcgtg cggccgcata ataaaccgat acaattaaag
     2581 gctccttttg gagccttttt tttggagatt ttcaacgtga aaaaattatt attcgcaatt
     2641 cctttagttg ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa
     2701 ccccatacag aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac
     2761 gctaactatg agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa
     2821 actcagtgtc tagctagagt ggcggtggct ctggttccgg tgattttgat tatgaaaaga
     2881 tggcaaacgc taataagggg gctatgaccg aaaatgccga tgaaaacgcg ctacagtctg
     2941 acgctaaagg caaacttgat tctgtcgcta ctgattacgg tgctgctatc gatggtttca
     3001 ttggtgacgt ttccggcctt gctaatggta atggtgctac tggtgatttt gctggctcta
     3061 attcccaaat ggctcaagtc ggtgacggtg ataattcacc tttaatgaat aatttccgtc
     3121 aatatttacc ttccctccct caatcggttg aatgtcgccc ttttgtcttt ggcgctggta
     3181 aaccatatga attttctatt gattgtgaca aaataaactt attccgtggt gtctttgcgt
     3241 ttcttttata tgttgccacc tttatgtatg tattttctac gtttgctaac atactgcgta
     3301 ataaggagtc ttaatcatgc cagttctttt ggctagcgcc gccctatacc ttgtctgcct
     3361 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     3421 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     3481 gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc
     3541 cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
     3601 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     3661 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     3721 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     3781 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     3841 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     3901 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     3961 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     4021 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     4081 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     4141 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcaggatc
     4201 cgcgaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4261 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4321 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4381 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4441 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4501 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4561 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4621 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cgcggatcct gcctcgcgcg
     4681 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4741 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4801 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     4861 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4921 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     4981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccagggtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5701 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5821 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5881 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5941 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6001 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6061 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6121 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6181 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     6241 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6301 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6361 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6421 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6481 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     6541 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6601 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6661 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6721 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6781 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6841 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6901 ttattatcat gacattaacc tataaaaata ggc
//
