LOCUS       sequence-207847-        5180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression expression of protein with N-terminal 
            lanthanide binding tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5180)
  AUTHORS   Bhattacharya K, Bernasconi L, Picard D
  TITLE     Luminescence resonance energy transfer between genetically encoded 
            donor and acceptor for protein-protein interaction studies in the 
            molecular chaperone HSP70/HSP90 complexes.
  JOURNAL   Sci Rep. 2018 Feb 12;8(1):2801. doi: 10.1038/s41598-018-21210-6.
  PUBMED    29434293
REFERENCE   2  (bases 1 to 5180)
  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"; journalName: "Sci Rep."; 
            date: "2018-02-12"; volume: "8(1):2801. doi"; pages: " 
            10.1038/s41598-018-21210-6"
COMMENT     Sequence Label: pST96/LBT-MCS
FEATURES             Location/Qualifiers
     source          1..5180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    60..84
                     /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             99..121
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             285..305
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             309..335
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(388..406)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      402..449
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      486..941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(573..592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     783..804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1033..1848)
                     /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(1033..1848)
                     /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     1756..1775
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1840..1859
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1970..2558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2459..2478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2712..2729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2744..2883
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2809..2828)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2969..2991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2985..3176)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3748..4830)
                     /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     4792..4811
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4831..4908)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5114..5133
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg
       61 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag
      121 atatacatat ggccatgggc tatatagata caaacaatga tggatggtat gaaggagatg
      181 aactattagc agctggaccg gtcgccatcg aattctgtac aggccttggc gcgcctgcag
      241 gcgagctccg tcgacaagct tgcggccgca ctcgagcctc cgcggagaat ctttattttc
      301 agggctccca tcaccatcac catcaccatc accattgacc taggctgcta aacaaagccc
      361 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      421 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
      481 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      541 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      601 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      661 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
      721 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      781 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      841 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      901 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
      961 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1021 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     1081 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1141 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1201 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1261 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     1321 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     1381 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     1441 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     1501 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     1561 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     1621 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     1681 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     1741 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     1801 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     1861 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     1921 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1981 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2041 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2101 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2161 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2221 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2281 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2341 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2401 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2461 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2521 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2581 tacggttcct ggccttttgc tggccttttg ctcgcatgtt ctttcctgcg ttatcccctg
     2641 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2701 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     2761 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     2821 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     2881 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2941 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3001 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     3061 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     3121 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     3181 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     3241 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     3301 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     3361 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     3421 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     3481 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     3541 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     3601 agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag
     3661 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     3721 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3781 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3841 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3901 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     3961 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4021 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4081 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4141 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4201 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4261 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4321 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4381 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4441 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4501 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4561 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4621 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4681 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     4741 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     4801 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4861 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     4921 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     4981 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5041 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5101 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5161 tgatgccggc cacgatgcgt
//
