LOCUS       sequence-357273-        5256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CoilE genetically encoded tag on H2B-mEmerald, to be used with CoilR
            labeled with a reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  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 5256)
  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: H2B-CoilE-mEm
FEATURES             Location/Qualifiers
     source          1..5256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             85..879
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             85..879
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(139..158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     749..768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1067..1086)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1111..1158
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1487..2075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1976..1995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2250..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2554..2757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2708..2728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2846..3223
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      3386..3395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3392..3394,3395..3397,3398..4111)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3437..3458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3698..3717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4045..4066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4233..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4270..4289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4324..4343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4361..4816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4498..4519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4710..4729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4843..4947
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4949..5256,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4949..5140
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4972..4992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5157..5256,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
       61 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      121 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      181 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      241 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
      301 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      361 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      421 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      481 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      541 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      601 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      661 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
      721 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
      781 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
      841 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
      901 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
      961 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     1021 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     1081 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     1141 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     1201 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     1261 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     1321 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1621 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2161 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2761 ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa
     2821 ttctgcagtc gacggtaccg ccaccatgcc agagccagcg aagtctgctc ccgccccgaa
     2881 aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg
     2941 cagccgcaag gagagctatt ccatctatgt gtacaaggtt ctgaagcagg tccaccctga
     3001 caccggcatt tcgtccaagg ccatgggcat catgaattcg tttgtgaacg acattttcga
     3061 gcgcatcgca ggtgaggctt cccgcctggc gcattacaac aagcgctcga ccatcacctc
     3121 cagggagatc cagacggccg tgcgcctgct gctgcctggg gagttggcca agcacgccgt
     3181 gtccgagggt actaaggcca tcaccaagta caccagcgct aaggatccac cggtaatgct
     3241 ggaaattgaa gcggcgtttc tggaacgtga gaataccgcg ctggaaaccc gtgtggcgga
     3301 actgcgtcag cgtgtgcagc gtctgcgtaa tcgtgtgagc cagtatcgta cccgttatgg
     3361 cccgctgggt ggcggcggac cggtcgccac catggtgagc aagggcgagg agctgttcac
     3421 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3481 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3541 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttgacct acggcgtgca
     3601 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3661 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3721 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3781 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3841 ggtctatatc accgccgaca agcagaagaa cggcatcaag gtgaacttca agacccgcca
     3901 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3961 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     4021 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4081 cactctcggc atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc
     4141 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4201 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4261 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4321 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4381 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4441 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4501 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4561 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4621 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4681 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4741 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4801 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4861 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4921 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4981 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5041 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5101 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5161 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5221 tttatgcaga ggccgaggcc gcctcggcct ctgagc
//
