LOCUS       sequence-192601-        6906 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            AC104650.1-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  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 6906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_AC104650.1-1/1
FEATURES             Location/Qualifiers
     source          1..6906
                     /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      2538..2585
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4178..4633)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4315..4336)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4527..4546
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4675..4697)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4783..4923
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4838..4857
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4938..4955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5109..5696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5867..6658,6659..6727))
                     /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     6490..6509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6728..6832)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(6900..6906,1..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 ccggtctggc atctgcactg gcagattttc aaggttccgg tccggaagaa atgactttcc
     2401 gtggtggtga tctgattgaa attctgggtg cccaagttcc atctctgcca tggtgtgttg
     2461 gtcgtcatgc agcatctggt cgtgttggtt ttgttcgttc ctccctgatt tccatgcaag
     2521 gtgcggccgc ataataaacc gatacaatta aaggctcctt ttggagcctt ttttttggag
     2581 attttcaacg tgaaaaaatt attattcgca attcctttag ttgttccttt ctattctcac
     2641 tccgctgaaa ctgttgaaag ttgtttagca aaaccccata cagaaaattc atttactaac
     2701 gtctggaaag acgacaaaac tttagatcgt tacgctaact atgagggctg tctgtggaat
     2761 gctacaggcg ttgtagtttg tactggtgac gaaactcagt gtctagctag agtggcggtg
     2821 gctctggttc cggtgatttt gattatgaaa agatggcaaa cgctaataag ggggctatga
     2881 ccgaaaatgc cgatgaaaac gcgctacagt ctgacgctaa aggcaaactt gattctgtcg
     2941 ctactgatta cggtgctgct atcgatggtt tcattggtga cgtttccggc cttgctaatg
     3001 gtaatggtgc tactggtgat tttgctggct ctaattccca aatggctcaa gtcggtgacg
     3061 gtgataattc acctttaatg aataatttcc gtcaatattt accttccctc cctcaatcgg
     3121 ttgaatgtcg cccttttgtc tttggcgctg gtaaaccata tgaattttct attgattgtg
     3181 acaaaataaa cttattccgt ggtgtctttg cgtttctttt atatgttgcc acctttatgt
     3241 atgtattttc tacgtttgct aacatactgc gtaataagga gtcttaatca tgccagttct
     3301 tttggctagc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     3361 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     3421 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     3481 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     3541 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     3601 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     3661 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     3721 tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc
     3781 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat
     3841 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
     3901 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
     3961 cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg
     4021 accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg
     4081 cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt
     4141 cacgaccacg ctgatgagct ttaccgcagg atccgcgaaa ttgtaagcgt taatattttg
     4201 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4261 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4321 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4381 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4441 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4501 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4561 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4621 ctacagggcg cgtcgcggat cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     4681 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4741 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     4801 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     4861 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4921 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4981 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5041 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5101 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5161 cagaggtggc gaaacccgac aggactataa agataccagg gtttccccct ggaagctccc
     5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5521 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6181 ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6481 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6841 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6901 ataggc
//
