LOCUS       sequence-190991-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            TEC-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  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_TEC-1/1
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     607..624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    639..779
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(705..724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     865..887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      929..1384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1016..1035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1226..1247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(2977..3024)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(3232..3249)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(3262..3285)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(3292..3945)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(3312..3334)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(3961..3978)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4052..4074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4063..4079
                     /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(4087..4093,4094..4109,4110..4115))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(4358..5440)
                     /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     5402..5421
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5441..5518)
                     /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(5541..5559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5627..5731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5732..5800,5801..6592)
                     /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(5950..5969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6763..6897,1..453)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
       61 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      121 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      181 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      241 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      301 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      361 ccctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      541 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      601 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
      661 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
      721 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
      781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      901 caccgaaacg cgcgaggcag gatccgcgac gcgccctgta gcggcgcatt aagcgcggcg
      961 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1021 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1081 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1141 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1201 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1261 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1321 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1381 atttcgcgga tcctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     1441 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     1501 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     1561 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     1621 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     1681 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     1741 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     1801 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     1861 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     1921 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     1981 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc
     2041 cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc
     2101 gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta
     2161 gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca
     2221 acgcggggag gcagacaagg tatagggcgg cgctagccaa aagaactggc atgattaaga
     2281 ctccttatta cgcagtatgt tagcaaacgt agaaaataca tacataaagg tggcaacata
     2341 taaaagaaac gcaaagacac cacggaataa gtttattttg tcacaatcaa tagaaaattc
     2401 atatggttta ccagcgccaa agacaaaagg gcgacattca accgattgag ggagggaagg
     2461 taaatattga cggaaattat tcattaaagg tgaattatca ccgtcaccga cttgagccat
     2521 ttgggaatta gagccagcaa aatcaccagt agcaccatta ccattagcaa ggccggaaac
     2581 gtcaccaatg aaaccatcga tagcagcacc gtaatcagta gcgacagaat caagtttgcc
     2641 tttagcgtca gactgtagcg cgttttcatc ggcattttcg gtcatagccc ccttattagc
     2701 gtttgccatc ttttcataat caaaatcacc ggaaccagag ccaccgccac tctagctaga
     2761 cactgagttt cgtcaccagt acaaactaca acgcctgtag cattccacag acagccctca
     2821 tagttagcgt aacgatctaa agttttgtcg tctttccaga cgttagtaaa tgaattttct
     2881 gtatggggtt ttgctaaaca actttcaaca gtttcagcgg agtgagaata gaaaggaaca
     2941 actaaaggaa ttgcgaataa taattttttc acgttgaaaa tctccaaaaa aaaggctcca
     3001 aaaggagcct ttaattgtat cggtttatta tgcggccgct tttttgccgg taacataatt
     3061 gctcggaatg taaccttcgt tgccatattt gtcgcgcgca cgccaccaat gcacatcgtt
     3121 tttttccagg atcagatatt cttgaccacg ttccagacgc agatcatgac cttcggcagc
     3181 ttgaaaatcg tacatagcga caacaatttc ggccggctgg gccgcggtac cggaaccacg
     3241 cggaaccaga ccgctggaca ggggcccctg gaacagaact tccagatccg attttggagg
     3301 atggtcgcca ccaccaaacg tggcttgcca gccctgcaaa ggccatgcta tatacttgct
     3361 ggatttcaag tacttatcaa tttgtgggat agcttcaata cgttttttaa aacaaactaa
     3421 ttttgggaac gcatccaggc acattgggtc catgtataaa acaacatcaa gagcgtcata
     3481 caacatgaag tcaggatggg ttacatgatc accatttaaa tatgttttat gacataaacg
     3541 atcttcgaac attttcagca tttcaggtag cttgctaaga aaatcaactt tgagagtttc
     3601 aaagtcttta ctatatgcaa ttctcgaaac accgtatcta atatccaaaa ccgctccttc
     3661 aagcattgaa atctctgcac gctcttttgg acaaccaccc aacatgttgt gcttgtcagc
     3721 tatataacgt atgatggcca tagactgtgt taatttaaca tcaccatcaa tataataagg
     3781 aagattggga aactccaaac ccaattcaaa ctttttgttt cgccatttat caccttcatc
     3841 gcgctcatac aaatgctctt catatttttc ttcaagatat tccaaaagaa gtcgagtggg
     3901 ttgcacaagg cccttaattt tccaataacc tagtataggg gacataagct taccgctgga
     3961 atggtgatgg tgatggtgtt cttcgatttt cataatctat ggtccttgtt ggtgaagtgc
     4021 tcgtgaaaac acctaaacgg actggctgtt tcctgtgtga aattgttatc cgctcacaat
     4081 tccacacatt atacgagccg atgattaatt gtcaacagct catttcagaa tatttgccag
     4141 aaccgttatg atgtcggcgc aaaaaacatt atccagaacg ggagtgcgcc ttgagcgaca
     4201 cgaattatgc agtgatttac gacctgcaca gccataccac agcttccgat ggctgcctga
     4261 cgccagaagc attggtgcac cgtgcagtcg atgataagct gtcaaacatg agaattgtgc
     4321 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4441 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4501 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4561 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4621 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4681 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4741 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4801 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4861 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     4921 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     4981 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5041 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5101 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5161 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5221 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5281 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5341 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5401 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5461 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcgg
     5521 gaattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5581 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5701 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5821 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5881 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5941 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     6001 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6061 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6121 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6181 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6241 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     6301 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6421 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6541 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6601 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagat
//
