LOCUS       sequence-306135-        6561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein fused to mRNA decapping enzyme 1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6561)
  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 6561)
  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: GFP-Dcp1b
FEATURES             Location/Qualifiers
     source          1..6561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        403..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     557..577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      645..654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             651..1367
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    3387..3508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3424..3443)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3478..3497
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3515..3970)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3652..3673)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3864..3883
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4103..4460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4103..4294
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4126..4146)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4311..4446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4495..5289
                     /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             4495..5289
                     /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(4549..4568)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5159..5178
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5477..5496)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5521..5568
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5897..6485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6386..6405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      601 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca
      661 agggcgccga gctgttcacc ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga
      721 atggccacaa gttcagcgtg agcggcgagg gcgagggcga tgccacctac ggcaagctga
      781 ccctgaagtt catctgcacc accggcaagc tgcctgtgcc ctggcccacc ctggtgacca
      841 ccctgagcta cggcgtgcag tgcttctcac gctaccccga tcacatgaag cagcacgact
      901 tcttcaagag cgccatgcct gagggctaca tccaggagcg caccatcttc ttcgaggatg
      961 acggcaacta caagtcgcgc gccgaggtga agttcgaggg cgataccctg gtgaatcgca
     1021 tcgagctgac cggcaccgat ttcaaggagg atggcaacat cctgggcaat aagatggagt
     1081 acaactacaa cgcccacaat gtgtacatca tgaccgacaa ggccaagaat ggcatcaagg
     1141 tgaacttcaa gatccgccac aacatcgagg atggcagcgt gcagctggcc gaccactacc
     1201 agcagaatac ccccatcggc gatggccctg tgctgctgcc cgataaccac tacctgtcca
     1261 cccagagcgc cctgtccaag gaccccaacg agaagcgcga tcacatgatc tacttcggct
     1321 tcgtgaccgc cgccgccatc acccacggca tggatgagct gtacaagtcc ggactcagat
     1381 ctcgagctgc agccgtggcg gcaggcggcc tggtgggaaa ggggcgcgac atcagcctag
     1441 cggccctgca gcgccacgac ccctatatca accgcatcgt ggacgtggcc agccaggtgg
     1501 ctctgtacac cttcggccat cgggccaacg agtgggagaa aactgatgtg gaaggaacct
     1561 tatttgttta tacaaggtct gcttctccaa agcatggatt caccattatg aataggctga
     1621 gcatggaaaa taggacagaa cctattacta aagacttgga tttccaactc caggaccctt
     1681 tccttctcta cagaaatgcc agattgtcca tctatggaat ttggttttat gataaggaag
     1741 aatgccaaag aattgcagag cttatgaaaa acctaactca gtatgaacag ttgaaagccc
     1801 atcagggaac tggagcagga atttcaccag tgatcctcaa ttcaggagag ggcaaagaag
     1861 tagacatttt acgaatgctc atcaaggcca aagacgaata cacaaagtgt aaaacctgtt
     1921 ctgagccaaa aaagataacc agttcctctg ccatctatga caatccaaat ctcatcaaac
     1981 caattccagt gaaacccagt gaaaaccagc aacagcgtat acctcagccc aaccagacct
     2041 tagaccctga accccaacac ttatccttga cagctctgtt tgggaagcag gacaaagcta
     2101 catgtcagga aactgtggag cctccgcaga ctctccacca gcagcagcag cagcagcagc
     2161 agcagcagca agagaagctt ccaattaggc agggggttgt acgctccctg tcctatgagg
     2221 aacccagaag acactcaccc cccattgaga agcagctctg tccagccatt cagaaactca
     2281 tggtcaggag cgcagacctc cacccattgt cagagctgcc tgaaaaccgg ccttgtgaaa
     2341 atggcagtac ccattctgcg ggagaatttt ttacaggacc tgtccagcca gggtctcctc
     2401 acaacattgg aacttctcgt ggtgtacaaa atgcttccag aactcagaac ctgttcgaga
     2461 aacttcagag taccccaggg gcagcaaaca agtgtgaccc tagtacacca gcacctgcca
     2521 gctcagctgc cctgaaccgc agcagagctc ccacttctgt cacccctgtg gctccaggaa
     2581 agggtctggc tcagccacca caggcctatt tcaatggctc ccttccacct cagacagtag
     2641 gacatcaggc tcatggaaga gaacagtcca cactcccaag acaaacactc gccatctctg
     2701 gtagtcagac tggcagctct ggagtgatct cccctcaaga gttactgaag aagcttcaga
     2761 ttgtacagca ggagcagcag ctgcatgcct ctaaccggcc agccttggcc gctaagtttc
     2821 ctgtgctcgc tcagagctct ggaacaggga aacccttgga atcctggatc aacaagacac
     2881 ccaacacaga acagcagact cctcttttcc aggtcatctc acctcagcgc atccctgcta
     2941 cagcagctcc gtctctgctt atgtccccca tggtgttcgc acaacccacc tccgtcccgc
     3001 caaaggaaag ggagagcggc ctcttgcctg tgggaggcca ggagccacct gttgctgcca
     3061 ccagcctcct cctgcccata cagagcccgg agccctccgt gatcaccagc agcccactca
     3121 ccaagctcca gctccaggag gcactgctgt acctcattca gaatgatgac aacttcttaa
     3181 atataatcta tgaagcctat ctcttcagca tgactcaagc agccatgaaa aagactatgt
     3241 gaggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     3301 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3361 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3481 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3541 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3601 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3661 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3721 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3781 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3841 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3901 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3961 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4141 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4381 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4441 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4501 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4561 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4621 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4681 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4741 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4801 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4861 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4921 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4981 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5041 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5101 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5161 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5221 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5281 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5341 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5401 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5461 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5521 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5581 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5641 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5701 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5761 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5821 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5881 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5941 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6001 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6061 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6121 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6181 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6241 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6301 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6361 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6421 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6481 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6541 acatgttctt tcctgcgtta t
//
