LOCUS       sequence-254704-        8129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CytERM-mCherry mRNA reporter tagged with (F30-2xPepper)10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8129)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins.
  JOURNAL   Nat Methods. 2019 Sep;16(9):862-865. doi: 10.1038/s41592-019-0531-7.
            Epub 2019 Aug 30.
  PUBMED    31471614
REFERENCE   2  (bases 1 to 8129)
  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: 
            "10.1038/s41592-019-0531-7"; journalName: "Nat Methods"; date: 
            "2019-09"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     Sequence Label: CMV-CytERM-mCherry-(F30-2xPepper)10
FEATURES             Location/Qualifiers
     source          1..8129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      677..686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             812..1519
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(954..972)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1222..1241
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3486..3503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3492..3716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3762..4190
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3849..3868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4059..4080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4199..4219)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4204..4533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4384..4519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4446..4465
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4600..5394
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4654..4673)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5264..5283
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5568..5689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5605..5624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5659..5678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5738..5754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5738..5754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5751..5773)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5762..5778
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5786..5792,5793..5810,5811..5816))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5831..5852
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5969..5986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6140..6728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6220..6239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6899..7690,7691..7759))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7522..7541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7760..7864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7939..7958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttgcca ccatggaccc tgtggtggtg ctggggctct gtctctcctg
      721 tttgcttctc ctttcactct ggaaacagag ctatggggga gggaaactgg gcggaagcgg
      781 agggacgggg ggttcaggaa cttcaggggg tgtgagcaag ggcgaggagg ataacatggc
      841 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
      901 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
      961 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     1021 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     1081 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1141 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     1201 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     1261 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     1321 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     1381 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     1441 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     1501 ggacgagctg tacaagtaac tcgagatccg ttacggccgg aatcaatcgc taatcactca
     1561 acttgccatg tgtatgtggg aagcgtagaa aggctcgttg agctcattag ctccgagccc
     1621 gactacgttt cccacatact ctgatgatcc gctagcaaag gctcgtctga gctcattagc
     1681 tccgagcccg aggtaccgga tcattcatgg caagtccagc gcaatctatt acgaaaatca
     1741 tccgacgtcg cgatgtctat gcgggaagcg tagaaaggct cgtctgagct cattagctcc
     1801 gagcccgact acgtttcccg catagtctga tcatccgcta gcaaaggctc gttgagctca
     1861 ttagctccga gcccgaggta ccggatgatt catcgcgacg ctgcggaaaa tctcacaaaa
     1921 tcacgtcaaa cgtcgccgtg tgtgtgtagg aagcgtagaa aggctcgtct gagctcatta
     1981 gctccgagcc cgactacgtt tcctacacac tctgacgatc cgctagcaaa ggctcgttga
     2041 gctcattagc tccgagcccg aggtaccgga tcgttcacgg cgacgccgat aatccacata
     2101 cttacaatca ggcaatcttg ccatgtgtat gtgggaagcg tagaaaggct cgttgagctc
     2161 attagctccg agcccgacta cgtttcccac atactctgat gatccgctag caaaggctcg
     2221 ttgagctcat tagctccgag cccgaggtac cggatcattc atggcaagta tcaagatcga
     2281 acggcgcaag atattgtcac gtcgcgatgt ctatgcggga agcgtagaaa ggctcgttga
     2341 gctcattagc tccgagcccg actacgtttc ccgcatagtc tgatcatccg ctagcaaagg
     2401 ctcgtctgag ctcattagct ccgagcccga ggtaccggat gattcatcgc gacgtcctcg
     2461 ctagatatgt taggttctta ggcatttcgc cgtgtgtgtg taggaagcgt agaaaggctc
     2521 gttgagctca ttagctccga gcccgactac gtttcctaca cactctgacg atccgctagc
     2581 aaaggctcgt ctgagctcat tagctccgag cccgaggtac cggatcgttc acggcgaaaa
     2641 gatcgtctgc aattccgatt agacgtacac ttgccatgtg tatgtgggaa gcgtagaaag
     2701 gctcgtctga gctcattagc tccgagcccg actacgtttc ccacatactc tgatgatccg
     2761 ctagcaaagg ctcgttgagc tcattagctc cgagcccgag gtaccggatc attcatggca
     2821 agatccaagc tacttcctcc atacctatcc tcctcgcgat gtctatgcgg gaagcgtaga
     2881 aaggctcgtc tgagctcatt agctccgagc ccgactacgt ttcccgcata gtctgatcat
     2941 ccgctagcaa aggctcgttg agctcattag ctccgagccc gaggtaccgg atgattcatc
     3001 gcgagatcat aacgcaatac cgtacactgt ccaatcctcg ccgtgtgtgt gtaggaagcg
     3061 tagaaaggct cgtctgagct cattagctcc gagcccgact acgtttccta cacactctga
     3121 cgatccgcta gcaaaggctc gttgagctca ttagctccga gcccgaggta ccggatcgtt
     3181 cacggcgagg ataatcaatc cacatacatc acaccacaat tcttgccatg tgtatgtggg
     3241 aagcgtagaa aggctcgtct gagctcatta gctccgagcc cgactacgtt tcccacatac
     3301 tctgatgatc cgctagcaaa ggctcgtctg agctcattag ctccgagccc gaggtaccgg
     3361 atcattcatg gcaagaattg gtcgttcttc ttggcggccg ctcgactaaa tcaccggtaa
     3421 tcttcttgtc catctagacc ttataaagat ctttgtacaa gggcccgttt aaacccgctg
     3481 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3541 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3601 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3661 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3721 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3781 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3841 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3901 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3961 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4021 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4081 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4141 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4201 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4261 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4321 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4381 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4441 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4501 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4561 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4621 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4681 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4741 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4801 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4861 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4921 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4981 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5041 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5101 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5161 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5221 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5281 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5341 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5401 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5461 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5521 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5581 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5641 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5701 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5761 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5821 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5881 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5941 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6001 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6061 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7861 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7921 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7981 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     8041 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     8101 tcgcgatgta cgggccagat atacgcgtt
//
