LOCUS       sequence-430149-        6385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a thermostable maltose binding protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6385)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 6385)
  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"
COMMENT     Sequence Label: Pfu-MBP
FEATURES             Location/Qualifiers
     source          1..6385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     216..235
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        216..234
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    235..259
                     /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)."
     RBS             274..296
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1461..1478
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1531..1549)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1545..1592
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1629..2084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1716..1735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1926..1947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2177..2992)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2177..2992)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2900..2919
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2984..3003
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3114..3702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3603..3622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3856..3873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3888..4030
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3956..3975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4116..4138
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4132..4323)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5132..6214)
                     /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     6176..6195
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6215..6292)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
       61 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
      121 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      181 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      241 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      301 ccatgggcat gaagatcgag gaaggtaagg ttgttatttg gcacgcgatg cagccgaacg
      361 agctggaagt gtttcaaagc ctggcggagg aatacatggc gctgtgcccg gaagtggaaa
      421 tcgttttcga acagaagccg aacctggagg acgcgctgaa agcggcgatt ccgaccggtc
      481 aaggtccgga cctgtttatc tgggcgcacg attggattgg caaattcgcg gaagcgggtc
      541 tgctggagcc gatcgacgaa tatgttaccg aggatctgct gaacgagttt gcgccgatgg
      601 cgcaggatgc gatgcaatac aagggccact actatgcgct gccgtttgcg gcggaaaccg
      661 tggcgatcat ttataacaag gaaatggtta gcgagccgcc gaaaaccttt gacgaaatga
      721 aggcgatcat ggagaaatac tatgatccgg cgaacgagaa atacggtatc gcgtggccga
      781 ttaacgcgta tttcatcagc gcgattgcgc aggcgtttgg tggctactat ttcgacgata
      841 agaccgaaca accgggcctg gacaaaccgg agaccatcga aggttttaag ttctttttca
      901 ccgagatttg gccgtacatg gcgccgaccg gcgattataa cacccagcaa agcatcttcc
      961 tggaaggccg tgcgccgatg atggtgaacg gtccgtggag catcaacgac gttaagaaag
     1021 cgggcattaa ctttggtgtg gttccgctgc cgccgatcat taaggatggc aaagagtact
     1081 ggccgcgtcc gtatggtggc gtgaaactga tctactttgc ggcgggtatt aagaacaaag
     1141 acgcggcgtg gaagttcgcg aaatggctga ccaccagcga ggaaagcatc aagaccctgg
     1201 cgctggaact gggttatatt ccggtgctga ccaaagttct ggacgatccg gagatcaaga
     1261 acgacccggt gatttacggc ttcggtcagg cggttcaaca cgcgtatctg atgccgaaga
     1321 gcccgaaaat gagcgcggtg tggggtggcg ttgatggtgc gatcaacgaa attctgcagg
     1381 acccgcaaaa cgcggacatc gaaggtattc tgaagaaata ccagcaagag atcctgaaca
     1441 acatgcagta ataactcgag caccaccacc accaccactg agatccggct gctaacaaag
     1501 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1561 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     1621 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1681 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1921 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1981 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2041 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcaggtg gcacttttcg
     2101 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2161 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     2221 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     2281 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     2341 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     2401 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     2461 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2521 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     2581 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     2641 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     2701 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2761 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2821 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2881 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2941 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     3001 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     3061 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3121 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3181 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3241 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3301 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3361 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3421 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3481 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3541 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3601 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3661 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3721 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3781 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3841 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3901 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     3961 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4021 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4081 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4141 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4201 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4261 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4321 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     4381 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4441 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4501 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     4561 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     4621 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     4681 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     4741 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     4801 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     4861 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     4921 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     4981 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     5041 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     5101 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5221 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     5281 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     5341 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     5401 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     5461 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     5521 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     5581 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     5641 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     5701 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     5761 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     5821 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     5881 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     5941 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     6001 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     6061 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     6121 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     6181 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     6241 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     6301 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     6361 ttgagcaccg ccgccgcaag gaatg
//
