LOCUS       sequence-257395-        5769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  AUTHORS   Rizzo MA, Davidson MW, Piston DW
  TITLE     Fluorescent protein tracking and detection: fluorescent protein 
            structure and color variants.
  JOURNAL   Cold Spring Harb Protoc. 2009 Dec;2009(12):pdb.top63. doi: 
            10.1101/pdb.top63.
  PUBMED    20150100
REFERENCE   2  (bases 1 to 5769)
  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: "Cold Spring
            Harb Protoc."; date: "2009-12"; volume: "2009(12):pdb.top63. doi"; 
            pages: " 10.1101/pdb.top63"
COMMENT     Sequence Label: mCherry-Actin-C-18
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        344..547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     498..518
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      586..595
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             592..1299
                     /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(737..755)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1005..1024
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2608..2729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2645..2664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2699..2718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2736..3191)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2873..2894)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3085..3104
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3218..3322
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3347..3367)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3391..3601
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3413..3609
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3460..3595
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3644..4438
                     /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             3644..4438
                     /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(3698..3717)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4308..4327
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4626..4645)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4670..4717
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5046..5634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5535..5554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
       61 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      301 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      361 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      421 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      481 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      541 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc
      601 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
      661 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      721 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
      781 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
      841 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      901 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
      961 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     1021 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     1081 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     1141 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     1201 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     1261 ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggactcag atctggcagc
     1321 ggtggaggca gcgcatccgg cggaagcgga agcgatgatg atatcgccgc gctcgtcgtc
     1381 gacaacggct ccggcatgtg caaggccggc ttcgcgggcg acgatgcccc ccgggccgtc
     1441 ttcccctcca tcgtggggcg ccccaggcac cagggcgtga tggtgggcat gggtcagaag
     1501 gattcctatg tgggcgacga ggcccagagc aagagaggca tcctcaccct gaagtacccc
     1561 atcgagcacg gcatcgtcac caactgggac gacatggaga aaatctggca ccacaccttc
     1621 tacaatgagc tgcgtgtggc tcccgaggag caccccgtgc tgctgaccga ggcccccctg
     1681 aaccccaagg ccaaccgcga gaagatgacc cagatcatgt ttgagacctt caacacccca
     1741 gccatgtacg ttgctatcca ggctgtgcta tccctgtacg cctctggccg taccactggc
     1801 atcgtgatgg actccggtga cggggtcacc cacactgtgc ccatctacga ggggtatgcc
     1861 ctcccccatg ccatcctgcg tctggacctg gctggccggg acctgactga ctacctcatg
     1921 aagatcctca ccgagcgcgg ctacagcttc accaccacgg ccgagcggga aatcgtgcgt
     1981 gacattaagg agaagctgtg ctacgtcgcc ctggacttcg agcaagagat ggccacggct
     2041 gcttccagct cctccctgga gaagagctac gagctgcctg acggccaggt catcaccatt
     2101 ggcaatgagc ggttccgctg ccctgaggca ctcttccagc cttccttcct gggcatggag
     2161 tcctgtggca tccacgaaac taccttcaac tccatcatga agtgtgacgt ggacatccgc
     2221 aaagacctgt acgccaacac agtgctgtct ggcggcacca ccatgtaccc tggcattgcc
     2281 gacaggatgc agaaggagat cactgccctg gcacccagca caatgaagat caagatcatt
     2341 gctcctcctg agcgcaagta ctccgtgtgg atcggcggct ccatcctggc ctcgctgtcc
     2401 accttccagc agatgtggat cagcaagcag gagtatgacg agtccggccc ctccatcgtc
     2461 caccgcaaat gcttctaggg atccaccgga tctagataac tgatcataat cagccatacc
     2521 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2581 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2641 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2701 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2761 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2821 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2881 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2941 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3001 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3061 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3121 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3181 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3241 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3301 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3361 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3421 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3481 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3541 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3601 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3661 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3721 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3781 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3841 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3901 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3961 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4021 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4081 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4141 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4201 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4261 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4321 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4381 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4441 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4501 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4561 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4621 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4681 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4741 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4801 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4861 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5641 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5701 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     5761 gtaatcaat
//
