LOCUS       sequence-191064-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            FUT8-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  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 6900)
  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_FUT8-1/1
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..24
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        47..124
                     /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             125..1207
                     /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(144..163)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1450..1455,1456..1471,1472..1478)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1486..1502
                     /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     1491..1513
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1587..1604
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1620..2273
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2231..2253
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2280..2303
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2316..2333
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2544..2591
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4184..4639)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4321..4342)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4533..4552
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4681..4703)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4789..4929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4844..4863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4944..4961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5115..5702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5873..6664,6665..6733))
                     /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     6496..6515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6734..6838)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attcccgaca ccatcgaatg
       61 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
      121 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
      181 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
      241 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
      301 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
      361 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
      421 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
      481 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
      541 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
      601 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
      661 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
      721 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
      781 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
      841 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
      901 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
      961 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     1021 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     1081 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     1141 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1201 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcacaa ttctcatgtt
     1261 tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga
     1321 agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     1381 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     1441 aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac
     1501 aatttcacac aggaaacagc cagtccgttt aggtgttttc acgagcactt caccaacaag
     1561 gaccatagat tatgaaaatc gaagaacacc atcaccatca ccattccagc ggtaagctta
     1621 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
     1681 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
     1741 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
     1801 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
     1861 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     1921 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     1981 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     2041 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     2101 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     2161 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     2221 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
     2281 tggaagttct gttccagggg cccctgtcca gcggtctggt tccgcgtggt tccggtaccg
     2341 cggcccagcc ggcccacaat cagatcgcta tttatgcaca tcagccgcgc accgcagacg
     2401 aaatcccgat ggaaccgggt gacattattg gcgtggcagg caaccattgg gatggctata
     2461 gcaaaggtgt gaatcgtaaa ctgggccgca ccggtctgta tccgtcttac aaagttcgtg
     2521 aaaaaattgc ggccgcataa taaaccgata caattaaagg ctccttttgg agcctttttt
     2581 ttggagattt tcaacgtgaa aaaattatta ttcgcaattc ctttagttgt tcctttctat
     2641 tctcactccg ctgaaactgt tgaaagttgt ttagcaaaac cccatacaga aaattcattt
     2701 actaacgtct ggaaagacga caaaacttta gatcgttacg ctaactatga gggctgtctg
     2761 tggaatgcta caggcgttgt agtttgtact ggtgacgaaa ctcagtgtct agctagagtg
     2821 gcggtggctc tggttccggt gattttgatt atgaaaagat ggcaaacgct aataaggggg
     2881 ctatgaccga aaatgccgat gaaaacgcgc tacagtctga cgctaaaggc aaacttgatt
     2941 ctgtcgctac tgattacggt gctgctatcg atggtttcat tggtgacgtt tccggccttg
     3001 ctaatggtaa tggtgctact ggtgattttg ctggctctaa ttcccaaatg gctcaagtcg
     3061 gtgacggtga taattcacct ttaatgaata atttccgtca atatttacct tccctccctc
     3121 aatcggttga atgtcgccct tttgtctttg gcgctggtaa accatatgaa ttttctattg
     3181 attgtgacaa aataaactta ttccgtggtg tctttgcgtt tcttttatat gttgccacct
     3241 ttatgtatgt attttctacg tttgctaaca tactgcgtaa taaggagtct taatcatgcc
     3301 agttcttttg gctagcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
     3361 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
     3421 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
     3481 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
     3541 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
     3601 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
     3661 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
     3721 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc
     3781 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
     3841 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta
     3901 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc
     3961 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca
     4021 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca
     4081 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa
     4141 tcgcttcacg accacgctga tgagctttac cgcaggatcc gcgaaattgt aagcgttaat
     4201 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4261 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4321 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4381 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4441 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4501 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4561 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4621 gcgccgctac agggcgcgtc gcggatcctg cctcgcgcgt ttcggtgatg acggtgaaaa
     4681 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4741 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     4801 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     4861 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4921 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accagggttt ccccctggaa
     5221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5521 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6181 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6481 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6841 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
//
