LOCUS       sequence-191073-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            DNMBP-3/6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  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 6894)
  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_DNMBP-3/6
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..31
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        54..131
                     /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             132..1214
                     /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(151..170)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1457..1462,1463..1478,1479..1485)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1493..1509
                     /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     1498..1520
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1594..1611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1627..2280
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2238..2260
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2287..2310
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2323..2340
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2545..2592
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4185..4640)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4322..4343)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4534..4553
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4682..4704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4790..4930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4845..4864
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4945..4962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5116..5703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5874..6665,6666..6734))
                     /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     6497..6516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6735..6839)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt cccgacacca
       61 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
      121 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
      181 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
      241 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
      301 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
      361 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
      421 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
      481 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
      541 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
      601 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
      661 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
      721 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
      781 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
      841 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
      901 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
      961 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     1021 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     1081 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     1141 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1201 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc
     1261 tcatgtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     1321 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     1381 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     1441 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
     1501 ggataacaat ttcacacagg aaacagccag tccgtttagg tgttttcacg agcacttcac
     1561 caacaaggac catagattat gaaaatcgaa gaacaccatc accatcacca ttccagcggt
     1621 aagcttatgt cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     1681 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     1741 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     1801 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     1861 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     1921 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     1981 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     2041 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     2101 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     2161 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     2221 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     2281 tcggatctgg aagttctgtt ccaggggccc ctgtccagcg gtctggttcc gcgtggttcc
     2341 ggtaccgcgg cccagccggc cggtcaagcc cgtgctctga tgggtctgtc ggcgcaactg
     2401 gatgaagaac tggatttccg tgaaggcgat gtgattacga ttattggcgt gccggaaccg
     2461 ggctggtttg aaggtgaact ggaaggccgt cgcggtattt ttccggaagg cttcgttgaa
     2521 ctgctgggtg cggccgcata ataaaccgat acaattaaag gctccttttg gagccttttt
     2581 tttggagatt ttcaacgtga aaaaattatt attcgcaatt cctttagttg ttcctttcta
     2641 ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa ccccatacag aaaattcatt
     2701 tactaacgtc tggaaagacg acaaaacttt agatcgttac gctaactatg agggctgtct
     2761 gtggaatgct acaggcgttg tagtttgtac tggtgacgaa actcagtgtc tagctagagt
     2821 ggcggtggct ctggttccgg tgattttgat tatgaaaaga tggcaaacgc taataagggg
     2881 gctatgaccg aaaatgccga tgaaaacgcg ctacagtctg acgctaaagg caaacttgat
     2941 tctgtcgcta ctgattacgg tgctgctatc gatggtttca ttggtgacgt ttccggcctt
     3001 gctaatggta atggtgctac tggtgatttt gctggctcta attcccaaat ggctcaagtc
     3061 ggtgacggtg ataattcacc tttaatgaat aatttccgtc aatatttacc ttccctccct
     3121 caatcggttg aatgtcgccc ttttgtcttt ggcgctggta aaccatatga attttctatt
     3181 gattgtgaca aaataaactt attccgtggt gtctttgcgt ttcttttata tgttgccacc
     3241 tttatgtatg tattttctac gtttgctaac atactgcgta ataaggagtc ttaatcatgc
     3301 cagttctttt ggctagcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
     3361 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
     3421 cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc
     3481 ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg
     3541 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     3601 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     3661 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     3721 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     3781 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     3841 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     3901 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     3961 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     4021 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     4081 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     4141 atcgcttcac gaccacgctg atgagcttta ccgcaggatc cgcgaaattg taagcgttaa
     4201 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4261 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4321 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4381 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4441 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4501 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4561 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4621 tgcgccgcta cagggcgcgt cgcggatcct gcctcgcgcg tttcggtgat gacggtgaaa
     4681 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4741 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     4801 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     4861 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4921 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4981 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5041 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5101 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5161 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccagggtt tccccctgga
     5221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5521 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6181 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6481 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6841 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gaca
//
