LOCUS       sequence-388063-        3721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Thermal stability of BPTI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3721)
  AUTHORS   Islam MM, Kobayashi K, Kidokoro SI, Kuroda Y
  TITLE     Hydrophobic surface residues can stabilize a protein through 
            improved water-protein interactions.
  JOURNAL   FEBS J. 2019 Oct;286(20):4122-4134. doi: 10.1111/febs.14941. Epub 
            2019 Jun 17.
  PUBMED    31175706
REFERENCE   2  (bases 1 to 3721)
  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: 
            "10.1111/febs.14941"; journalName: "FEBS J"; date: "2019-10"; 
            volume: "286"; issue: "20"; pages: "4122-4134"
COMMENT     Sequence Label: BPTI-[5,55]Gly14Ile38
FEATURES             Location/Qualifiers
     source          1..3721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             84..110
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(701..719)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      715..762
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1008..1025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1179..1767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1259..1278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1938..2729,2730..2798))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2561..2580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2799..2903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2971..2989
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3027..3049)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3149..3168
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3186..3699)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3402)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3593..3612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcatcacc atcaccatca ccatcaccat aaagcaattt
      121 tcgtactgaa aggttcactg gacagagatc tcgacagccg tattgaactg gaactgcgta
      181 ccgatcataa agagctgtct gaacatctgc tgctggttga tctcgcccgt aatgatctgg
      241 cacgcattgc tacccccggc agccgctacg tcgccgatct caccaaagtt gaccgttatt
      301 cctatgtgct gcacctcgtc tctcgcgtag tcggcgaact gcgtcacgat cttgacgccc
      361 tgcacgctta tcgcgccgct ctgaatctgg ggacgttaag cggtgcgccg aaagtacgcg
      421 ctaagctttg gatgcgtccg gacttctgcc tcgagccgcc gtacaccggt ccgggcaaag
      481 cgcgcatcat ccgttacttc tacaacgcca aggcaggcct ggctcagacc ttcgtatacg
      541 gtggtatccg tgctaagcgt aacaacttca aatccgcgga agacgctctt cgtacctgcg
      601 gtggtgcttg aggatccccg ggtaccgagc tcgaattcat cgatgatatc agatccggct
      661 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      721 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      781 tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag
      841 tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg
      901 tgcgcataga aattgcatca acgcatatag cgctagagtc gactgcatta atgaatcggc
      961 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1021 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1081 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1141 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1201 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1261 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1321 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1381 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1441 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1501 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1561 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1621 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1681 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1741 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1801 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1861 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1921 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1981 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2041 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2101 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2161 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2221 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2281 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2341 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2401 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2461 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2521 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2581 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2641 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2701 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2761 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2821 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2881 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2941 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     3001 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3061 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3121 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3181 catccacccc ggttgataat cagaaaagcc ccaaaaacag gaagattgta taagcaaata
     3241 tttaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3361 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatctcga tcccgcgaaa
     3721 t
//
