LOCUS       sequence-192424-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            MPP4-1/1-IS3/3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  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 6972)
  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_MPP4-1/1-IS3/3
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..29
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        52..129
                     /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             130..1212
                     /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(149..168)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1455..1460,1461..1476,1477..1483)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1491..1507
                     /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     1496..1518
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1592..1609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1625..2278
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2236..2258
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2285..2308
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2321..2338
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2621..2668
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4261..4716)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4398..4419)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4610..4629
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4758..4780)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4866..5006
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4921..4940
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5021..5038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5192..5779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5950..6741,6742..6810))
                     /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     6573..6592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6811..6915)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattcc cgacaccatc
       61 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
      121 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
      181 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
      241 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
      301 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
      361 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
      421 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
      481 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
      541 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
      601 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
      661 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
      721 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
      781 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
      841 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
      901 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
      961 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     1021 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     1081 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     1141 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1201 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg cacaattctc
     1261 atgtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     1321 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     1381 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     1441 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
     1501 ataacaattt cacacaggaa acagccagtc cgtttaggtg ttttcacgag cacttcacca
     1561 acaaggacca tagattatga aaatcgaaga acaccatcac catcaccatt ccagcggtaa
     1621 gcttatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact
     1681 tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga
     1741 taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat
     1801 tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca
     1861 caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg aaggagcggt
     1921 tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg aaactctcaa
     1981 agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca
     2041 taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct
     2101 tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt
     2161 taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat
     2221 agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc
     2281 ggatctggaa gttctgttcc aggggcccct gtccagcggt ctggttccgc gtggttccgg
     2341 taccgcggcc cagccggcca tggcccaggg cgcgctgtct cgtggtacta tcatgttcaa
     2401 agttgtccca gtttacgatc caccagttaa ctctcaacag atggacccag atattccatg
     2461 tatggatgcc ggtctgccat ttcagaaagg tgatattctg caaattgttg atcagaacga
     2521 tgcactgtgg tggcaagcac gtaaaatttc tgatccagcc acctgtgcag gtctggttcc
     2581 atctaaccat ctgctgaaac gtaaagcggc cgcataataa accgatacaa ttaaaggctc
     2641 cttttggagc cttttttttg gagattttca acgtgaaaaa attattattc gcaattcctt
     2701 tagttgttcc tttctattct cactccgctg aaactgttga aagttgttta gcaaaacccc
     2761 atacagaaaa ttcatttact aacgtctgga aagacgacaa aactttagat cgttacgcta
     2821 actatgaggg ctgtctgtgg aatgctacag gcgttgtagt ttgtactggt gacgaaactc
     2881 agtgtctagc tagagtggcg gtggctctgg ttccggtgat tttgattatg aaaagatggc
     2941 aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc
     3001 taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg gtttcattgg
     3061 tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg gctctaattc
     3121 ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt tccgtcaata
     3181 tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttggcg ctggtaaacc
     3241 atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct ttgcgtttct
     3301 tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac tgcgtaataa
     3361 ggagtcttaa tcatgccagt tcttttggct agcgccgccc tataccttgt ctgcctcccc
     3421 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     3481 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     3541 atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca
     3601 cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
     3661 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     3721 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     3781 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     3841 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     3901 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     3961 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     4021 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     4081 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     4141 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     4201 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc aggatccgcg
     4261 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4321 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4381 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4441 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4501 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4561 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4621 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4681 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg gatcctgcct cgcgcgtttc
     4741 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4801 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4861 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     4921 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     4981 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     5041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5281 agggtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5341 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5401 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5461 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5521 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5581 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5641 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5701 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6001 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6061 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6121 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6181 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6241 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     6301 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6361 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6421 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6481 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6541 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     6601 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6661 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6721 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6781 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6841 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6901 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6961 tatcatgaca tt
//
