LOCUS       sequence-213886-        7155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of STAM_VHS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7155)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 7155)
  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"; journalName: "Mol Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_STAM_VHS
FEATURES             Location/Qualifiers
     source          1..7155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      1350..1397
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(2990..3445)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3127..3148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3339..3358
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3487..3509)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3595..3735
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3650..3669
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3750..3767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3921..4508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4679..5470,5471..5539))
                     /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     5302..5321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5540..5644)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5712..5730
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5753..5830
                     /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             5831..6913
                     /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(5850..5869)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggccatgcc gctgttcgct actaaccctt tcgaccagga cgttgaaaaa gctacttccg
      961 agatgaacac cgcggaagac tggggcctga tcctggatat ctgcgacaag gtaggtcagt
     1021 ctcgcaccgg tccaaaagat tgtctgcgct ctattatgcg tcgtgttaac cataaagatc
     1081 cgcacgtggc gatgcaggcg ctgacgctgc tgggtgcctg cgtaagcaac tgcggcaaaa
     1141 tcttccacct ggaagtctgc tctcgtgatt ttgcatccga ggtttccaac gtgctgaaca
     1201 aaggtcaccc gaaagtttgt gaaaaactga aagctctgat ggtggaatgg accgatgaat
     1261 ttaagaacga cccgcaactg agcctgattt ctgcaatgat caagaatctg aaagaacagg
     1321 gcgttacctt cccggcggcc gcataataaa ccgatacaat taaaggctcc ttttggagcc
     1381 ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccttt agttgttcct
     1441 ttctattctc actccgctga aactgttgaa agttgtttag caaaacccca tacagaaaat
     1501 tcatttacta acgtctggaa agacgacaaa actttagatc gttacgctaa ctatgagggc
     1561 tgtctgtgga atgctacagg cgttgtagtt tgtactggtg acgaaactca gtgtctagct
     1621 agagtggcgg tggctctggt tccggtgatt ttgattatga aaagatggca aacgctaata
     1681 agggggctat gaccgaaaat gccgatgaaa acgcgctaca gtctgacgct aaaggcaaac
     1741 ttgattctgt cgctactgat tacggtgctg ctatcgatgg tttcattggt gacgtttccg
     1801 gccttgctaa tggtaatggt gctactggtg attttgctgg ctctaattcc caaatggctc
     1861 aagtcggtga cggtgataat tcacctttaa tgaataattt ccgtcaatat ttaccttccc
     1921 tccctcaatc ggttgaatgt cgcccttttg tctttggcgc tggtaaacca tatgaatttt
     1981 ctattgattg tgacaaaata aacttattcc gtggtgtctt tgcgtttctt ttatatgttg
     2041 ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag gagtcttaat
     2101 catgccagtt cttttggcta gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg
     2161 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga
     2221 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
     2281 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
     2341 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
     2401 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
     2461 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
     2521 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
     2581 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     2641 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
     2701 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
     2761 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
     2821 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     2881 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     2941 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca ggatccgcga aattgtaagc
     3001 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3061 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3121 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3181 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3241 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3301 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3361 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3421 cttaatgcgc cgctacaggg cgcgtcgcgg atcctgcctc gcgcgtttcg gtgatgacgg
     3481 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3541 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     3601 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     3661 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3721 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3901 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca gggtttcccc
     4021 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4081 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4141 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4201 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4261 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4321 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4381 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4441 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4501 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4561 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4621 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4981 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5281 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5641 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     5701 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc ccgacaccat
     5761 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     5821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     5881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     5941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     6001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     6061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     6121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     6181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     6241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     6301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     6361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     6421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     6481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     6541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     6601 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     6661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     6721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     6781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     6841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcacaattct
     6961 catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
     7021 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
     7081 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
     7141 ttctgaaatg agctg
//
