LOCUS       sequence-192361-        6923 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            DNMBP-5/6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6923)
  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 6923)
  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_DNMBP-5/6
FEATURES             Location/Qualifiers
     source          1..6923
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     26..44
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        67..144
                     /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             145..1227
                     /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(164..183)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1470..1475,1476..1491,1492..1498)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1506..1522
                     /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     1511..1533
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1607..1624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1640..2293
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2251..2273
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2300..2323
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2336..2353
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2588..2635
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4228..4682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4365..4386)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4576..4595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4724..4746)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4832..4972
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4887..4906
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4987..5004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5158..5745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5916..6707,6708..6776))
                     /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     6539..6558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6777..6881)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
       61 attcccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
      121 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      181 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      241 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      301 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      361 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      421 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      481 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      541 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      601 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      661 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      721 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      781 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      841 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      901 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      961 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     1021 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1081 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1141 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1201 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     1261 ttaggcacaa ttctcatgtt tgacagctta tcatcgactg cacggtgcac caatgcttct
     1321 ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa
     1381 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     1441 ggttctggca aatattctga aatgagctgt tgacaattaa tcatcggctc gtataatgtg
     1501 tggaattgtg agcggataac aatttcacac aggaaacagc cagtccgttt aggtgttttc
     1561 acgagcactt caccaacaag gaccatagat tatgaaaatc gaagaacacc atcaccatca
     1621 ccattccagc ggtaagctta tgtcccctat actaggttat tggaaaatta agggccttgt
     1681 gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga
     1741 gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa
     1801 tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta
     1861 tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat
     1921 gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga
     1981 ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga
     2041 agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat
     2101 gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc
     2161 aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa
     2221 atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga
     2281 ccatcctcca aaatcggatc tggaagttct gttccagggg cccctgtcca gcggtctggt
     2341 tccgcgtggt tccggtaccg cggcccagcc ggccatggcc cagggcgcgc tgaaactgtt
     2401 tcaggccgaa cgtaacttta acgcagccca ggatctggat gtttctctgc tggaaggtga
     2461 tctcgttggt gttatcaaaa agaaagatcc gatgggttcc caaaatcgtt ggctgattga
     2521 taacggtgtt accaaaggtt ttgtctattc ctccttcctg aaaccataca acgcggccgc
     2581 ataataaacc gatacaatta aaggctcctt ttggagcctt ttttttggag attttcaacg
     2641 tgaaaaaatt attattcgca attcctttag ttgttccttt ctattctcac tccgctgaaa
     2701 ctgttgaaag ttgtttagca aaaccccata cagaaaattc atttactaac gtctggaaag
     2761 acgacaaaac tttagatcgt tacgctaact atgagggctg tctgtggaat gctacaggcg
     2821 ttgtagtttg tactggtgac gaaactcagt gtctagctag agtggcggtg gctctggttc
     2881 cggtgatttt gattatgaaa agatggcaaa cgctaataag ggggctatga ccgaaaatgc
     2941 cgatgaaaac gcgctacagt ctgacgctaa aggcaaactt gattctgtcg ctactgatta
     3001 cggtgctgct atcgatggtt tcattggtga cgtttccggc cttgctaatg gtaatggtgc
     3061 tactggtgat tttgctggct ctaattccca aatggctcaa gtcggtgacg gtgataattc
     3121 acctttaatg aataatttcc gtcaatattt accttccctc cctcaatcgg ttgaatgtcg
     3181 cccttttgtc tttggcgctg gtaaaccata tgaattttct attgattgtg acaaaataaa
     3241 cttattccgt ggtgtctttg cgtttctttt atatgttgcc acctttatgt atgtattttc
     3301 tacgtttgct aacatactgc gtaataagga gtcttaatca tgccagttct tttggctagc
     3361 gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc
     3421 tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag
     3481 ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca
     3541 tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc
     3601 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     3661 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     3721 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     3781 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     3841 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     3901 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     3961 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     4021 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     4081 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     4141 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     4201 ctgatgagct ttaccgcagg atccgcgaaa ttgtaagcgt taatattttg ttaaaattcg
     4261 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4321 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4381 gtccactatt aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4441 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4501 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4561 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4621 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4681 gtcgcggatc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4741 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4801 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     4861 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4921 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     4981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5221 aaacccgaca ggactataaa gataccaggg tttccccctg gaagctccct cgtgcgctct
     5281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5581 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     6241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6301 cgagttacat gatcccccat attgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     6541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6901 ccacctgacg tctaagaaac cat
//
