LOCUS       sequence-430162-        6006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the full length of silica binding protein fused at the 
            N-terminus of large algae metallothionein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6006)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 6006)
  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-FvMT
FEATURES             Location/Qualifiers
     source          1..6006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     144..163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     397..414
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    429..568
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(494..513)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     654..676
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(670..861)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1670..2752)
                     /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     2714..2733
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2753..2830)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3036..3055
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3139..3158
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3139..3157
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3158..3182
                     /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             3197..3219
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4009..4026
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4079..4097)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4093..4140
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4177..4632
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4264..4283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4474..4495
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4724..5539)
                     /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(4724..5539)
                     /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     5447..5466
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5531..5550
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5661..6006,1..243)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
       61 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      301 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      361 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      421 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
      481 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
      541 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
      601 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
      661 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
      721 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
      781 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
      841 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
      901 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
      961 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     1021 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     1081 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     1141 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     1201 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     1261 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     1321 cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     1381 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     1441 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     1501 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     1561 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     1621 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     1681 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1741 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     1801 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     1861 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     1921 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     1981 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     2041 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     2101 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     2161 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     2221 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     2281 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     2341 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     2401 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     2461 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     2521 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     2581 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     2641 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     2701 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     2761 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     2821 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     2881 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     2941 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     3001 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     3061 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
     3121 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
     3181 cccctctaga aataattttg tttaacttta agaaggagat ataccatggc catgggtatg
     3241 tccctgttcc attgtgattt cctgaaagac ctgataggta gtttcgtccg cgtgaatcgc
     3301 ggtgggccag aaagccagcg cggaacaatt atctctgttt gtcccgatta ttttgtatta
     3361 caaaacgaga aaggcgaatt gtattactac cagctctctc atttaaagag tattacgaaa
     3421 agcgcaaaag aatgtggaag ctcggattgt gagtgggaag actgtgcttg cgcggaagac
     3481 ttcgaagcgc tcctcgaatc gttcaaatat tgttgggtaa agattaatcg aggcggtccc
     3541 gaaaaggtgg aaggaattct tcaggatgtc tcttgtgatt tcgttacact gattgtaaaa
     3601 gaggaaatca tcttgatagc gatcaaacat atcaaaagcg tcaactataa cgcactggct
     3661 tgtggggaaa gtgacgaaag tgatgacagc agctcaaaag aatctaacaa caactcaggg
     3721 cgcgcacgcg cccaacgaca atcatctcgc ggacgtggcg gtggtggctc gggaggaggc
     3781 ggtagcggta tggcaggcac tggttgcaag atttgggaag attgcaaatg tggggccgct
     3841 tgctcatgtg gggattcgtg tacgtgcggt actgttaaga agggcacaac tagcagagcc
     3901 ggggctggat gcccttgcgg acccaagtgt aaatgtacag gacaaggatc atgtaattgt
     3961 gtcaaggacg actgttgtgg atgtggaaaa tgataactcg agctcgagca ccaccaccac
     4021 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     4081 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     4141 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     4201 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4261 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4381 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4441 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4501 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4561 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4621 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4681 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     4741 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     4801 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     4861 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     4921 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     4981 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     5041 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     5101 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     5161 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     5221 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     5281 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     5341 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     5401 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     5461 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     5521 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     5581 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6001 cccagc
//
