LOCUS       sequence-414746-        5385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the His-tagged peptide PuRRRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5385)
  AUTHORS   Suyama A, Devlin KL, Macias-Contreras M, Doh JK, Shinde U, Beatty KE
  TITLE     Orthogonal Versatile Interacting Peptide Tags for Imaging Cellular 
            Proteins.
  JOURNAL   Biochemistry. 2023 Jun 6;62(11):1735-1743. doi: 
            10.1021/acs.biochem.2c00712. Epub 2023 May 11.
  PUBMED    37167569
REFERENCE   2  (bases 1 to 5385)
  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"; doi: 
            "10.1021/acs.biochem.2c00712"; journalName: "Biochemistry"; date: 
            "2023-06-6- 6"; volume: "62"; issue: "11"; pages: "1735-1743"
COMMENT     Sequence Label: pET28b-PuRRRE
FEATURES             Location/Qualifiers
     source          1..5385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     955..974
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(994..1071)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1277..1296
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1380..1399
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1380..1398
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1399..1423
                     /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             1438..1460
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1629..1646
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1699..1717)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1713..1760
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1797..2252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1884..1903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2094..2115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2344..3159)
                     /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(2344..3159)
                     /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     3067..3086
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3151..3170
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3281..3869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3770..3789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4023..4040
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4055..4194
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4120..4139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4280..4302
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4296..4487)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5296..5385,1..993))
                     /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 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
       61 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
      121 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
      181 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
      241 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
      301 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
      361 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
      421 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
      481 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
      541 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
      601 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
      661 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
      721 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
      781 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
      841 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
      901 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
      961 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1021 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     1081 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     1141 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     1201 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     1261 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     1321 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     1381 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     1441 gtttaacttt aagaaggaga tataccatgg gttcctctct ggaaatccgt aacgccttcc
     1501 tccgtcagcg tgttaccgcg ctgcgtaccc gcgtttctga actgcgtcag gaaaactccg
     1561 aactggaaaa cgaatacggt ccgctgggtt cttggggtct gtgctacccg tgggtttacg
     1621 gtctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1681 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1741 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     1801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1861 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1981 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2101 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2161 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2221 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     2281 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     2341 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2401 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2461 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2521 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2581 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     2641 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     2701 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     2761 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     2821 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     2881 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     2941 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3001 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3061 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3121 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     3181 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3241 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3301 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3361 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3421 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3481 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3541 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3601 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3661 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3721 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3781 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3841 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3901 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3961 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4021 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4081 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     4141 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4201 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4261 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4321 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4381 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4441 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4501 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4561 atacgggtta ctgatgatga acatgccngg ttactggaac gttgtgaggg taaacaactg
     4621 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4681 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4741 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4801 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4861 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4921 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     4981 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     5041 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     5101 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     5161 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     5221 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggg
//
