LOCUS       sequence-430163-        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the full length of silica binding protein fused at the 
            N-terminus of proteobacteria metallothionein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6027)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 6027)
  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: Si-ComMT
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     408..427
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(447..524)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     730..749
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     833..852
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        833..851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    852..876
                     /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             891..913
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1724..1741
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1794..1812)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1808..1855
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1892..2347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1979..1998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2189..2210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2439..3254)
                     /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(2439..3254)
                     /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     3162..3181
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3246..3265
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3376..3964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3865..3884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4118..4135
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4150..4289
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4215..4234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4375..4397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4391..4582)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5391..6027,1..446))
                     /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"
ORIGIN
        1 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
       61 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
      121 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
      181 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
      241 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
      301 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
      361 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
      421 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
      481 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
      541 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
      601 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
      661 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
      721 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
      781 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
      841 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      901 tttaagaagg agatatacca tggccatggg tatgagtctg ttccattgtg atttcttgaa
      961 agacttaatt ggcagcttcg tccgcgtcaa tagaggcggt cctgaatcgc agcgaggaac
     1021 tattatcagt gtgtgtcccg attattttgt attacaaaac gaaaagggag aactctatta
     1081 ctaccagctc agccatttga aatcaatcac gaaatctgcc aaagaatgtg gaagcagcga
     1141 ttgtgagtgg gaagactgtg cgtgcgctga ggacttcgaa gcgctgctgg aatctttcaa
     1201 atattgttgg gtaaagatta atcggggcgg gccggagaaa gtggaaggaa tcctgcagga
     1261 tgtttcctgt gatttcgtca ccttgattgt gaaagaggaa ataatcctta tagccattaa
     1321 acacataaaa tcagtgaact ataacgcact cgcatgtggc gaatcagacg aatcagatga
     1381 ctcatcttcc aaagaatcga acaacaactc tggaagagct cgtgcccaac gccagtcatc
     1441 tcgtgggaga ggcggtggcg ggagcggtgg cggcggatct ggtatgtacg accaacgatg
     1501 tgcctgtacg cactgttcct gtacggtcgt tgctaaggcc ttacaacgtg acggaaaggc
     1561 ctactgttgt gaggcttgtg ccagtggaca ccgtaaggga gagccttgtc gtatgcaaga
     1621 ctgtcactgt ggtgagaagc caggggagtc agctgttgac aatgcattag acgagacgtt
     1681 ccctgcatca gaccctatat caccatagta actcgagctc gagcaccacc accaccacca
     1741 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1801 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1861 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1921 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1981 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2041 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2101 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2161 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2221 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2281 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2341 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2401 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     2461 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     2521 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     2581 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     2641 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2701 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     2761 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     2821 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     2881 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     2941 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3001 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3061 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3121 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3181 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     3241 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     3301 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3361 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3421 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3481 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3541 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3601 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3661 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3721 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3781 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3841 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3901 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3961 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4021 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4081 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4141 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     4201 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     4261 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     4321 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4381 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     4441 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     4501 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4561 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     4621 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     4681 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     4741 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     4801 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     4861 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4921 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4981 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     5041 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     5101 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     5161 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     5221 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     5281 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     5341 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     5401 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5461 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     5521 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     5581 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     5641 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     5701 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     5761 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     5821 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     5881 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     5941 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     6001 ctgttgatgg gtgtctggtc agagaca
//
