LOCUS       sequence-357297-        5409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to label proteins with the CoilE genetically encoded tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5409)
  AUTHORS   Doh JK, White JD, Zane HK, Chang YH, Lopez CS, Enns CA, Beatty KE
  TITLE     VIPER is a genetically encoded peptide tag for fluorescence and 
            electron microscopy.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Dec 18;115(51):12961-12966. doi: 
            10.1073/pnas.1808626115. Epub 2018 Dec 5.
  PUBMED    30518560
REFERENCE   2  (bases 1 to 5409)
  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: 
            "10.1073/pnas.1808626115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-12-18- 18"; volume: "115"; issue: "51"; pages: 
            "12961-12966"
COMMENT     Sequence Label: pET28b-CoilR-Cys
FEATURES             Location/Qualifiers
     source          1..5409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     930..949
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(969..1046)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1252..1271
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1355..1374
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1355..1373
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1374..1398
                     /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             1413..1435
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1628..1645
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1698..1716)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1712..1759
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1796..2251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1883..1902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2093..2114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2343..3158)
                     /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(2343..3158)
                     /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     3066..3085
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3150..3169
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3280..3868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3769..3788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4022..4039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4054..4193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4119..4138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4279..4301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4295..4486)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5295..5409,1..968))
                     /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 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
       61 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
      121 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
      181 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
      241 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
      301 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
      361 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
      421 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
      481 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
      541 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
      601 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
      661 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
      721 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
      781 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
      841 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
      901 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
      961 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     1021 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     1081 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     1141 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     1201 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     1261 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     1321 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     1381 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     1441 catgggcggc agccttgaaa ttcgtgcggc ttttctgcgt cagcgtaata ccgcgctgcg
     1501 caccgaagtg gcggaactgg aacaggaagt gcagcgtctg gaaaatgaag tgagccagta
     1561 tgaaacccgt tatggcccgc tgggaggcgg agccgctgct ctgggctgtc ttgcggccgc
     1621 actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     1681 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1741 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1981 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2101 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2161 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2221 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     2281 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     2341 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2401 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2461 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2521 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2581 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2641 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2701 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2761 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2821 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     2881 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2941 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3001 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3061 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3121 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3181 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3421 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3901 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3961 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4021 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     4081 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     4141 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     4201 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4261 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4321 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4381 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4441 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4501 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4561 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4621 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4681 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4741 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4801 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4861 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4921 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     4981 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     5041 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     5101 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     5161 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     5221 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     5281 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     5401 cttttcacc
//
