LOCUS       sequence-306034-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SNAP-tag polypeptide fused to mRNA decapping enzyme 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  AUTHORS   Lee JE, Cathey PI, Wu H, Parker R, Voeltz GK
  TITLE     Endoplasmic reticulum contact sites regulate the dynamics of 
            membraneless organelles.
  JOURNAL   Science. 2020 Jan 31;367(6477). pii: 367/6477/eaay7108. doi: 
            10.1126/science.aay7108.
  PUBMED    32001628
REFERENCE   2  (bases 1 to 5787)
  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: "Science."; 
            date: "2020-01-31"; volume: "367(6477). pii: 367/6477/eaay7108. 
            doi"; pages: " 10.1126/science.aay7108"
COMMENT     Sequence Label: SNAP-Dcp2
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             249..794
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    2211..2332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2248..2267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2302..2321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2339..2794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2476..2497)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2688..2707
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2821..2925
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2927..3284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2927..3118
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2950..2970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3135..3270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3319..4113
                     /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             3319..4113
                     /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(3373..3392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3983..4002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4301..4320)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4345..4392
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4721..5309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5210..5229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5484..5787
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
      301 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
      361 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
      421 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
      481 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
      541 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
      601 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
      661 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac gtggggggct
      721 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
      781 agcctgggct gggtagcggt agcggtggca gctctcgagc tgagaccaaa cgggtggaga
      841 ttcccggcag cgtcctggac gatctctgca gccgatttat tttgcatatt cccagcgagg
      901 aaagagacaa tgcaatccga gtgtgttttc agattgaact tgcccattgg ttttacttgg
      961 atttctacat gcagaacaca ccaggattac ctcagtgtgg gataagagac tttgctaaag
     1021 ctgtcttcag tcattgtccg tttttgctgc ctcaaggtga agatgtggaa aaagttttgg
     1081 atgaatggaa ggaatataaa atgggagtac caacatatgg tgcaattatt cttgatgaga
     1141 cacttgaaaa tgtactactg gttcaggggt acctagcaaa atcaggctgg ggatttccaa
     1201 aaggaaaagt aaataaagaa gaagctcctc atgattgtgc tgctagagag gtctttgaag
     1261 aaactggttt tgatatcaaa gactatattt gtaaggatga ttacattgaa cttcgaatca
     1321 atgaccagct tgctcgtttg tacatcattc caggaattcc aaaagacaca aaatttaacc
     1381 caaaaactag aagagaaatt cggaacattg agtggttctc tattgagaaa ttgccttgtc
     1441 atagaaatga tatgaccccc aaatccaaac ttggtttggc acctaacaaa ttttttatgg
     1501 ccattccctt tatcagacca ttaagggact ggctttctcg aagatttggc gattcctcag
     1561 acagtgacaa tggattttcc tcaactggta gcacgccggc taaacccact gtggaaaaat
     1621 tgagtcgaac caaattccgc cacagtcagc agttatttcc tgacggttct cctggtgacc
     1681 agtgggtaaa gcacaggcaa ccactgcagc aaaagccata taataatcat tctgaaatgt
     1741 ctgacctttt aaaaggaaag aatcaaagta tgaggggaaa tggcagaaaa cagtatcaag
     1801 attcacctaa tcaaaagaaa agaacaaatg ggcttcagcc agcaaagcag cagaattctt
     1861 tgatgaagtg tgaaaagaaa cttcatccac ggaaacttca ggataatttt gaaacagatg
     1921 ctgtatatga cttgcctagc tccagtgaag accagttgct agaacatgcc gagggacagc
     1981 ccgtggcatg taatggacat tgcaagttcc ccttttcatc cagagccttt ttgagtttca
     2041 agtttgacca taatgctata atgaaaatct tggacctttg aggatccacc ggatctagat
     2101 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2161 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2221 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2281 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     2341 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2401 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2461 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2521 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2581 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2641 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2701 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2761 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2941 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3061 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3241 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3301 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3361 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3421 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3481 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3541 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3601 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3661 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3721 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3781 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3841 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3901 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3961 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4021 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4081 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4141 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4201 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4261 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4321 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4381 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4441 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4501 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4561 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4621 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4681 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4741 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4801 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4861 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4921 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4981 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5041 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5101 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5161 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5221 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5281 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5341 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5401 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     5461 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5521 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5581 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     5641 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     5701 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5761 tacgtattag tcatcgctat taccatg
//
