LOCUS       sequence-430161-        6015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the full length of silica binding protein fused at the 
            N-terminus of human metallothionein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6015)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 6015)
  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-rHMT
FEATURES             Location/Qualifiers
     source          1..6015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1726..1917
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1911..1933)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2019..2158
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2074..2093
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2173..2190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2344..2932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2424..2443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3043..3062)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3054..3869
                     /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             3054..3869
                     /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     complement(3127..3146)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3961..4416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4098..4119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4310..4329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4453..4500)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4496..4514
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4567..4584)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5383..5405
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5420..5444
                     /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     complement(5444..5463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5445..5463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5547..5566)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5772..5849
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5850..6015,1..917)
                     /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(5869..5888)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
       61 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
      121 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
      181 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
      241 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
      301 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
      361 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
      421 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
      481 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
      541 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
      601 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
      661 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
      721 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
      781 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
      841 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      901 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
      961 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1021 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     1081 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     1141 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     1201 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca
     1261 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     1321 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     1381 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     1441 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     1501 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     1561 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     1621 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     1681 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     1741 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     1801 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     1861 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     1921 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     1981 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     2041 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     2101 tgcaccattg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2761 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3001 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     3061 atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt
     3121 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     3181 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3241 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3301 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3361 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3421 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3481 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3541 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3601 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3661 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3721 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3781 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3841 ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa
     3901 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3961 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4021 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4081 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4141 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4201 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4261 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4321 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4381 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccaatc cggatatagt
     4441 tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag
     4501 ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg
     4561 atctcagtgg tggtggtggt ggtgctcgag ctcgagttat caagccgccg ccgctttctt
     4621 gcaacaggag cacttttcgc ttgctccttt acaaacgcaa ccctgtgcgc atttagcgca
     4681 tgacatcgga cagcacgagc aacacgcggc agctgctttc ttacaagaat tacacttaca
     4741 ttccttacat ttgcaggacc cggtgcatgt acaacttcca cccgttgcgc aactgcacgc
     4801 ggcagcagcc tttcctgacc cacctcctcc acttccgcca ccaccgcgac cacgcgatga
     4861 ttgtctttgt gcccgtgcgc gtcccgagtt attgttgctt tctttactgc tggagtcatc
     4921 tgattcgtcg ctttcaccac atgccagggc gttatagttg acggatttga tatgcttgat
     4981 tgcaatcaag attatttcct ctttcacaat aagggttacg aaatcacatg agacatcttg
     5041 cagaattcct tccacctttt caggcccgcc acgattaatc ttaacccaac aatatttgaa
     5101 agattccagc agggcttcga agtcctcggc gcacgcacag tcttcccact cacaatcgga
     5161 gctaccacat tctttagcac ttttcgtaat acttttcaaa tgagacagct ggtagtaata
     5221 caattcccct ttctcgtttt gcagcacaaa ataatccgga catacggaga taatagtgcc
     5281 tctctgcgat tctggcccac cacgattcac tcttacgaaa gatccaatta agtcttttaa
     5341 gaaatcacaa tggaacaagg acattcccat ggccatggta tatctccttc ttaaagttaa
     5401 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
     5461 ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac
     5521 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     5581 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     5641 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     5701 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     5761 tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     5821 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     5881 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     5941 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     6001 gcgtggcaca acaac
//
