LOCUS       sequence-357265-        5997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CoilE genetically encoded tag on mEmerald-Actin, 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 5997)
  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 5997)
  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: mEm-CoilE-Actin
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(628..630,631..633,634..1344)
                     /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(673..694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(934..953)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1281..1302
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2800..2921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2837..2856)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2891..2910
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2928..3383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3065..3086)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3277..3296
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3410..3514
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3516..3873
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3516..3707
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3539..3559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3724..3859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3908..4702
                     /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             3908..4702
                     /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(3962..3981)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4572..4591
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4890..4909)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4934..4981
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5310..5898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5799..5818
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      661 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      721 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      781 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tgacctacgg cgtgcagtgc
      841 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      901 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      961 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1021 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaaggtc
     1081 tatatcaccg ccgacaagca gaagaacggc atcaaggtga acttcaagac ccgccacaac
     1141 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1201 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     1261 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1321 ctcggcatgg acgagctgta caagtccgga ctcagatcta tgctggaaat tgaagcggcg
     1381 tttctggaac gtgagaatac cgcgctggaa acccgtgtgg cggaactgcg tcagcgtgtg
     1441 cagcgtctgc gtaatcgtgt gagccagtat cgtacccgtt atggcccgct gggtggcggc
     1501 agatctggca gcggtggagg cagcgcatcc ggcggaagcg gaagcgatga tgatatcgcc
     1561 gcgctcgtcg tcgacaacgg ctccggcatg tgcaaggccg gcttcgcggg cgacgatgcc
     1621 ccccgggccg tcttcccctc catcgtgggg cgccccaggc accagggcgt gatggtgggc
     1681 atgggtcaga aggattccta tgtgggcgac gaggcccaga gcaagagagg catcctcacc
     1741 ctgaagtacc ccatcgagca cggcatcgtc accaactggg acgacatgga gaaaatctgg
     1801 caccacacct tctacaatga gctgcgtgtg gctcccgagg agcaccccgt gctgctgacc
     1861 gaggcccccc tgaaccccaa ggccaaccgc gagaagatga cccagatcat gtttgagacc
     1921 ttcaacaccc cagccatgta cgttgctatc caggctgtgc tatccctgta cgcctctggc
     1981 cgtaccactg gcatcgtgat ggactccggt gacggggtca cccacactgt gcccatctac
     2041 gaggggtatg ccctccccca tgccatcctg cgtctggacc tggctggccg ggacctgact
     2101 gactacctca tgaagatcct caccgagcgc ggctacagct tcaccaccac ggccgagcgg
     2161 gaaatcgtgc gtgacattaa ggagaagctg tgctacgtcg ccctggactt cgagcaagag
     2221 atggccacgg ctgcttccag ctcctccctg gagaagagct acgagctgcc tgacggccag
     2281 gtcatcacca ttggcaatga gcggttccgc tgccctgagg cactcttcca gccttccttc
     2341 ctgggcatgg agtcctgtgg catccacgaa actaccttca actccatcat gaagtgtgac
     2401 gtggacatcc gcaaagacct gtacgccaac acagtgctgt ctggcggcac caccatgtac
     2461 cctggcattg ccgacaggat gcagaaggag atcactgccc tggcacccag cacaatgaag
     2521 atcaagatca ttgctcctcc tgagcgcaag tactccgtgt ggatcggcgg ctccatcctg
     2581 gcctcgctgt ccaccttcca gcagatgtgg atcagcaagc aggagtatga cgagtccggc
     2641 ccctccatcg tccaccgcaa atgcttctag ggatccaccg gatctagata actgatcata
     2701 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2761 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2821 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2881 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     2941 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3001 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3061 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3121 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3181 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3241 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3301 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3361 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3421 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3481 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3721 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3781 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3841 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3901 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3961 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4021 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4081 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4141 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4201 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4261 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4321 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4381 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4441 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4501 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4561 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4621 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4681 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4861 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4921 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4981 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5041 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5101 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5161 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5221 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5281 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5341 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5401 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5461 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5521 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5581 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5641 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5701 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5761 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5821 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5881 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5941 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgt
//
