LOCUS       sequence-497792-        5911 bp    DNA     circular SYN 20-JAN-2026
DEFINITION  mRNA-RELISR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5911)
  AUTHORS   Lee C, Yu J, Shin J, Yu J, Heo Y, Lee M, Yu D, Park Y, Heo WD
  TITLE     Optogenetic storage and release of protein and mRNA in live cells 
            and animals.
  JOURNAL   Nat Commun. 2025 Jul 7;16(1):6230. doi: 10.1038/s41467-025-61322-y.
  PUBMED    40624072
REFERENCE   2  (bases 1 to 5911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 20, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-07-7"; volume: "16(1):6230. doi"; pages: " 
            10.1038/s41467-025-61322-y"
COMMENT     Sequence Label: pCMV-tdMCP-DsRed-PixD
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1336..1345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1342..2016
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(1522..1542)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1740..1759
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1934..1957
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             complement(2051..2068)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2699..2820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2736..2755)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2790..2809
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2827..3282)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2964..2985)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3176..3195
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3309..3413
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3415..3772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3415..3606
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3438..3458)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3623..3758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3807..4601
                     /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             3807..4601
                     /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(3861..3880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4471..4490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4789..4808)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4833..4880
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5209..5797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5698..5717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccgtt
      601 aaaatggctt ctaactttac tcagttcgtt ctcgtcgaca atggcggaac tggcgacgtg
      661 actgtcgccc caagcaactt cgctaacggg atcgctgaat ggatcagctc taactcgcgt
      721 tcacaggctt acaaagtaac ctgtagcgtt cgtcagagct ctgcgcagaa tcgcaaatac
      781 accatcaaag tcgaggtgcc taaaggcgcc tggcgttcgt acttaaatat ggaactaacc
      841 attccaattt tcgccacgaa ttccgactgc gagcttattg ttaaggcaat gcaaggtctc
      901 ctaaaagatg gaaacccgat tccctcagca atcgcagcaa actccggcat ctacgccatg
      961 gcttctaact ttactcagtt cgttctcgtc gacaatggcg gaactggcga cgtgactgtc
     1021 gccccaagca acttcgctaa cgggatcgct gaatggatca gctctaactc gcgttcacag
     1081 gcttacaaag taacctgtag cgttcgtcag agctctgcgc agaatcgcaa atacaccatc
     1141 aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa atatggaact aaccattcca
     1201 attttcgcca cgaattccga ctgcgagctt attgttaagg caatgcaagg tctcctaaaa
     1261 gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgc ggattctaga
     1321 atggtgagac cggtcgccac catggatagc actgagaacg tcatcaagcc cttcatgcgc
     1381 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1441 gagggcaagc cctacgaggg cacccagacc gccaagctgc aggtgaccaa gggcggcccc
     1501 ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg
     1561 aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg
     1621 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1681 caggacggca ccttcatcta ccacgtgaag ttcatcggcg tgaacttccc ctccgacggc
     1741 cccgtaatgc agaagaagac tctgggctgg gagccctcca ccgagcgcct gtacccccgc
     1801 gacggcgtgc tgaagggcga gatccacaag gcgctgaagc tgaagggcgg cggccactac
     1861 ctggtggagt tcaagtcaat ctacatggcc aagaagcccg tgaagctgcc cggctactac
     1921 tacgtggact ccaagctgga catcacctcc cacaacgagg actacaccgt ggtggagcag
     1981 tacgagcgcg ccgaggcccg ccaccacctg ttccagttgt acagtggggg aggcggaggg
     2041 ggtggttcag gtggtggtgg tggtggtggt tcgggtggtg gcggaggggg aggctccgga
     2101 aaggccacgc gtatgagttt gtaccgtttg atttacagca gtcagggcat tcccaattta
     2161 caaccccagg atttaaaaga tatcttagaa tcttcccaaa gaaataatcc ggccaatggc
     2221 attaccggac tgctctgcta cagcaagccg gcttttctgc aggtattgga aggagagtgt
     2281 gaacaggtta atgaaaccta ccaccggatt gtgcaagatg aacgccacca cagcccccag
     2341 atcattgagt gtatgccgat cagaagaaga aacttcgagg tttggtctat gcaagcgatc
     2401 acggtgaatg atttgtcaac ggaacaggtt aagactttgg tattgaaata ttccggcttc
     2461 accaccctac ggcccagtgc catggacccg gaacaatgcc taaacttttt gctagacatt
     2521 gccaagattt acgagttgtc cgataacttt ttcctcgacc tctaagcggg gatccaccgg
     2581 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2641 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2701 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2761 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2821 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2881 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2941 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3001 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3061 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3121 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3181 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3241 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3301 gaaatgtgcg cggaacccct atttgtttat ttttttaaat acattcaaat atgtatccgc
     3361 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3421 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3481 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3541 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3601 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3661 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3721 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3781 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4621 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4681 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4741 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4801 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4861 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4921 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4981 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5041 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5101 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5161 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5221 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5281 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5341 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5401 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5461 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5521 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5581 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5641 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5701 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5761 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5821 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5881 ttctgtggat aaccgtatta ccgccatgca t
//
