LOCUS       sequence-248451-        8130 bp    DNA     circular SYN 14-APR-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 8130)
  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 8130)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..8130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      912..921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1047..1754
                     /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(1189..1207)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1457..1476
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3721..3738)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3727..3954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4000..4428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4087..4106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4297..4318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4437..4457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4442..4771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4622..4757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4684..4703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4838..5632
                     /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(4892..4911)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5502..5521
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5806..5927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5843..5862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5897..5916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5976..5992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5976..5992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5989..6011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6000..6016
                     /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(6024..6030,6031..6048,6049..6054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6069..6090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6207..6224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6378..6963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6458..6477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7134..7925,7926..7994))
                     /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     7757..7776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7995..8099)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact tgccaccatg gaccctgtgg tggtgctggg gctctgtctc tcctgtttgc
      961 ttctcctttc actctggaaa cagagctatg ggggagggaa actgggcgga agcggaggga
     1021 cggggggttc aggaacttca gggggtgtga gcaagggcga ggaggataac atggccatca
     1081 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     1141 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1201 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1261 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1321 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1381 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1441 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1501 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1561 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1621 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1681 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1741 agctgtacaa gtaactcgag atccgttacg gccggaatca atcgctaatc actcaacttg
     1801 ccatgtgtat gtgggaagcg tagaaaggct cgttgagctc attagctccg agcccgacta
     1861 cgtttcccac atactctgat gatccgctag caaaggctcg tctgagctca ttagctccga
     1921 gcccgaggta ccggatcatt catggcaagt ccagcgcaat ctattacgaa aatcatccga
     1981 cgtcgcgatg tctatgcggg aagcgtagaa aggctcgtct gagctcatta gctccgagcc
     2041 cgactacgtt tcccgcatag tctgatcatc cgctagcaaa ggctcgttga gctcattagc
     2101 tccgagcccg aggtaccgga tgattcatcg cgacgctgcg gaaaatctca caaaatcacg
     2161 tcaaacgtcg ccgtgtgtgt gtaggaagcg tagaaaggct cgtctgagct cattagctcc
     2221 gagcccgact acgtttccta cacactctga cgatccgcta gcaaaggctc gttgagctca
     2281 ttagctccga gcccgaggta ccggatcgtt cacggcgacg ccgataatcc acatacttac
     2341 aatcaggcaa tcttgccatg tgtatgtggg aagcgtagaa aggctcgttg agctcattag
     2401 ctccgagccc gactacgttt cccacatact ctgatgatcc gctagcaaag gctcgttgag
     2461 ctcattagct ccgagcccga ggtaccggat cattcatggc aagtatcaag atcgaacggc
     2521 gcaagatatt gtcacgtcgc gatgtctatg cgggaagcgt agaaaggctc gttgagctca
     2581 ttagctccga gcccgactac gtttcccgca tagtctgatc atccgctagc aaaggctcgt
     2641 ctgagctcat tagctccgag cccgaggtac cggatgattc atcgcgacgt cctcgctaga
     2701 tatgttaggt tcttaggcat ttcgccgtgt gtgtgtagga agcgtagaaa ggctcgttga
     2761 gctcattagc tccgagcccg actacgtttc ctacacactc tgacgatccg ctagcaaagg
     2821 ctcgtctgag ctcattagct ccgagcccga ggtaccggat cgttcacggc gaaaagatcg
     2881 tctgcaattc cgattagacg tacacttgcc atgtgtatgt gggaagcgta gaaaggctcg
     2941 tctgagctca ttagctccga gcccgactac gtttcccaca tactctgatg atccgctagc
     3001 aaaggctcgt tgagctcatt agctccgagc ccgaggtacc ggatcattca tggcaagatc
     3061 caagctactt cctccatacc tatcctcctc gcgatgtcta tgcgggaagc gtagaaaggc
     3121 tcgtctgagc tcattagctc cgagcccgac tacgtttccc gcatagtctg atcatccgct
     3181 agcaaaggct cgttgagctc attagctccg agcccgaggt accggatgat tcatcgcgag
     3241 atcataacgc aataccgtac actgtccaat cctcgccgtg tgtgtgtagg aagcgtagaa
     3301 aggctcgtct gagctcatta gctccgagcc cgactacgtt tcctacacac tctgacgatc
     3361 cgctagcaaa ggctcgttga gctcattagc tccgagcccg aggtaccgga tcgttcacgg
     3421 cgaggataat caatccacat acatcacacc acaattcttg ccatgtgtat gtgggaagcg
     3481 tagaaaggct cgtctgagct cattagctcc gagcccgact acgtttccca catactctga
     3541 tgatccgcta gcaaaggctc gtctgagctc attagctccg agcccgaggt accggatcat
     3601 tcatggcaag aattggtcgt tcttcttggc ggccgctcga ctaaatcacc ggtaatcttc
     3661 ttgtccatct agaccttata aagatctttg tacaagggcc cgtttaaacc cgctgatcag
     3721 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3781 tgaccctgga aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3841 cattgtctga gtaggtgtca ttctattctg gggggtgggg gtgggggcag gacagcaagg
     3901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3961 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4021 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4081 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4141 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4201 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4261 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4321 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4381 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4441 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4501 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4561 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4621 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4681 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4741 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4801 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4861 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4921 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4981 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5041 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5101 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5161 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5221 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5281 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5341 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5401 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5461 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5521 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5581 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5641 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5701 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5761 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5821 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5881 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5941 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6001 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6061 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6121 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6181 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6241 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6301 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6541 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6781 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6901 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6961 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7021 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7081 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7141 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7201 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7261 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7321 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7381 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7441 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7501 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7561 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7621 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7681 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7741 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7801 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7861 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7921 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7981 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8041 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8101 acatttcccc gaaaagtgcc acctgacgtc
//
