LOCUS       sequence-191823-        6918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            PACSIN3-1/1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  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 6918)
  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_PACSIN3-1/1
FEATURES             Location/Qualifiers
     source          1..6918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..21
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        44..121
                     /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             122..1204
                     /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(141..160)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1447..1452,1453..1468,1469..1475)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1483..1499
                     /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     1488..1510
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1584..1601
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1617..2270
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2228..2250
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2277..2300
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2313..2330
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2559..2606
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4199..4654)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4336..4357)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4548..4567
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4696..4718)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4804..4944
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4859..4878
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4959..4976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5130..5717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5888..6679,6680..6748))
                     /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     6511..6530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6749..6853)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt cccgacacca tcgaatggtg
       61 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
      121 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
      181 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
      241 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
      301 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
      361 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
      421 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
      481 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
      541 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
      601 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
      661 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
      721 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
      781 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
      841 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
      901 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
      961 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     1021 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     1081 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     1141 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     1201 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc tcatgtttga
     1261 cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     1321 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
     1381 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
     1441 gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat
     1501 ttcacacagg aaacagccag tccgtttagg tgttttcacg agcacttcac caacaaggac
     1561 catagattat gaaaatcgaa gaacaccatc accatcacca ttccagcggt aagcttatgt
     1621 cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg
     1681 aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc
     1741 gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg
     1801 atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt
     1861 tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata
     1921 ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt
     1981 ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat
     2041 atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg
     2101 ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac
     2161 gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc
     2221 ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg
     2281 aagttctgtt ccaggggccc ctgtccagcg gtctggttcc gcgtggttcc ggtaccgcgg
     2341 cccagccggc catggcccag ggcgcgctgg ttcgtgttcg tgccctgtat gattatgcag
     2401 gtcaggaagc agatgaactg tcttttcgtg ccggtgaaga actgctgaaa atgtccgaag
     2461 aagatgaaca aggctggtgt caaggtcaac tgcaatccgg tcgtattggt ctctatccag
     2521 ccaactatgt tgaatgcgtc ggtgcggccg cataataaac cgatacaatt aaaggctcct
     2581 tttggagcct tttttttgga gattttcaac gtgaaaaaat tattattcgc aattccttta
     2641 gttgttcctt tctattctca ctccgctgaa actgttgaaa gttgtttagc aaaaccccat
     2701 acagaaaatt catttactaa cgtctggaaa gacgacaaaa ctttagatcg ttacgctaac
     2761 tatgagggct gtctgtggaa tgctacaggc gttgtagttt gtactggtga cgaaactcag
     2821 tgtctagcta gagtggcggt ggctctggtt ccggtgattt tgattatgaa aagatggcaa
     2881 acgctaataa gggggctatg accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta
     2941 aaggcaaact tgattctgtc gctactgatt acggtgctgc tatcgatggt ttcattggtg
     3001 acgtttccgg ccttgctaat ggtaatggtg ctactggtga ttttgctggc tctaattccc
     3061 aaatggctca agtcggtgac ggtgataatt cacctttaat gaataatttc cgtcaatatt
     3121 taccttccct ccctcaatcg gttgaatgtc gcccttttgt ctttggcgct ggtaaaccat
     3181 atgaattttc tattgattgt gacaaaataa acttattccg tggtgtcttt gcgtttcttt
     3241 tatatgttgc cacctttatg tatgtatttt ctacgtttgc taacatactg cgtaataagg
     3301 agtcttaatc atgccagttc ttttggctag cgccgcccta taccttgtct gcctccccgc
     3361 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     3421 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     3481 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     3541 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     3601 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     3661 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     3721 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     3781 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     3841 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     3901 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     3961 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     4021 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     4081 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4141 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag gatccgcgaa
     4201 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4261 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4321 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4381 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4441 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4501 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4561 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4621 cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgga tcctgcctcg cgcgtttcgg
     4681 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4741 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4801 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     4861 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4921 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5221 ggtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5761 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5821 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5881 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5941 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6001 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6061 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6181 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atattgtgca
     6301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6481 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     6541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6841 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6901 tcatgacatt aacctata
//
