LOCUS       sequence-367107-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry labeled with 10xPP7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  AUTHORS   Guo Y, Lee REC
  TITLE     Long-term imaging of individual mRNA molecules in living cells.
  JOURNAL   Cell Rep Methods. 2022 May 25;2(6):100226. doi: 
            10.1016/j.crmeth.2022.100226. eCollection 2022 Jun 20.
  PUBMED    35784652
REFERENCE   2  (bases 1 to 6534)
  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: "Cell Rep 
            Methods."; date: "2022-05-25"; volume: "2(6):100226. doi"; pages: " 
            10.1016/j.crmeth.2022.100226. eCollection 2022 Jun 20"
COMMENT     Sequence Label: cmv-mCherry-10xPP7
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(121..140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      934..1522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1423..1442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1676..1693
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1810..1831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1846..1851,1852..1869,1870..1876)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1884..1900
                     /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)."
     primer_bind     1889..1911
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1908..1924
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1908..1924
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1943..1963
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1945..1963
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(2014..2047)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            complement(2110..3466)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(2110..2307)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2313..3122)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3036..3055
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3123..3466)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      complement(3536..3723)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     3644..3668
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(3749..3782)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4362..5072)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4640..4659)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4909..4927
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5069..5078
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5100..5303)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5129..5149)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5304..5607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5664..5683)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5665..5683)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5693..5710)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5693..5709)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5702..5724)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5851..6306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5938..5957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6148..6169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6332..6436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6437..6505,6506..6534,1..763)
                     /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"
ORIGIN
        1 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
       61 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      181 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      301 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      481 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      721 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
      781 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
      841 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1081 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1261 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1321 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1381 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1441 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1501 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1561 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1621 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     1681 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1741 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1801 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     1861 cccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     1921 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctgggtac
     1981 cgcataggcc actagtggat ctgatatcac ctaataactt cgtatagcat acattatacg
     2041 aagttatatt aagggttctc gagagctcgt tttcgacact ggatggcggc gttagtatcg
     2101 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
     2161 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
     2221 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
     2281 gaaaacaaga atctttttat tgtcagtact gattagaaaa actcatcgag catcaaatga
     2341 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2401 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2461 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2521 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
     2581 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2641 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     2701 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2761 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttg
     2821 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     2881 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     2941 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3001 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3061 aaatcagcat ccatgttgga atttaatcgc ggcctcgaaa cgtgagtctt ttccttaccc
     3121 atggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa
     3181 gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc tacaggggcg
     3241 cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg caggtctgca
     3301 gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg
     3361 attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat gcccctgagc
     3421 tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg gccgggtgac
     3481 ccggcgggga cgaggcaagc taaacagatc caccctgtta tccctagcgg atctgccggt
     3541 agaggtgtgg tcaataagag cgacctcata ctatacctga gaaagcaacc tgacctacag
     3601 gaaagagtta ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt
     3661 gtatacactt atttttttta taacttattt aataataaaa atcataaatc ataagaaatt
     3721 cgcttattta gaagtggcag atcacctaat aacttcgtat agcatacatt atacgaagtt
     3781 atattaaggg ttagatcttc atgtctgctc gaagcggaag cttaggggga attcttccta
     3841 cgggtttcca tataaactcc gtagccagga cgtgacgttt ctgtttcgca ttccatataa
     3901 tgtcgaaact gcgagttagt acattgggta accatattta ctccaatctt aggaaagcgg
     3961 tgaataagcc attccttata atgtgcttac gcaagcgttg catactgcga agcctgatgt
     4021 acggtcacca tattgactcg taacagacag gaattggaga cccaatccat aaagggattt
     4081 catataatct ccttttgttc tcggcttcaa ggcgacggct tttctaaggg tttccatata
     4141 aactccttag tcacgaccag caggcgattc cgtttccata taaactggaa tcatgggtaa
     4201 aaacaagtgg gccgacggta aggtagcatt gtctcgcaga gtttcccata tgaaatctct
     4261 ggtcacgacc agcaggcgta tgggtttcca tataaactcc atatgtaaga attcgtcaac
     4321 gcgacgctag ataggagctc ggatctgacg gttcataaac cttacttgta cagctcgtcc
     4381 atgccgccgg tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc
     4441 tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg
     4501 ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc
     4561 ttcagcctct gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag
     4621 gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg
     4681 ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg
     4741 gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag
     4801 gacagcttca agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac
     4861 atgaactgag gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc
     4921 agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac
     4981 tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg
     5041 gccatgttat cctcctcgcc cttgctcacc atggtggcga ccggatcctt cactaaacga
     5101 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     5161 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     5221 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     5281 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     5341 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     5401 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     5461 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     5521 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     5581 gggccattta ccgtaagtta tgtaacgcgg aactccacta gtccaaactg gagcggccgc
     5641 caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     5701 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5761 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5821 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     5881 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5941 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     6001 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     6061 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6121 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6181 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6241 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     6301 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6361 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6421 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6481 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aaga
//
