LOCUS       sequence-470324-        5969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 5969)
  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"
COMMENT     Sequence Label: mCherry-Calreticulin-N-16
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1858..1867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1864..2574
                     /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(2009..2027)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2277..2296
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2697..2818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2734..2753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2788..2807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2825..3280)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2962..2983)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3174..3193
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3307..3411
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3413..3770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3413..3604
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3436..3456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3621..3756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3805..4599
                     /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             3805..4599
                     /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(3859..3878)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4469..4488
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4787..4806)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4831..4878
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5207..5795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5696..5715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgaggc caccatgctg ctatccgtgc cgctgctgct cggcctcctc
      601 ggcctggccg tcgccgagcc tgccgtctac ttcaaggagc agtttctgga cggagacggg
      661 tggacttccc gctggatcga atccaaacac aagtcagatt ttggcaaatt cgttctcagt
      721 tccggcaagt tctacggtga cgaggagaaa gataaaggtt tgcagacaag ccaggatgca
      781 cgcttttatg ctctgtcggc cagtttcgag cctttcagca acaaaggcca gacgctggtg
      841 gtgcagttca cggtgaaaca tgagcagaac atcgactgtg ggggcggcta tgtgaagctg
      901 tttcctaata gtttggacca gacagacatg cacggagact cagaatacaa catcatgttt
      961 ggtcccgaca tctgtggccc tggcaccaag aaggttcatg tcatcttcaa ctacaagggc
     1021 aagaacgtgc tgatcaacaa ggacatccgt tgcaaggatg atgagtttac acacctgtac
     1081 acactgattg tgcggccaga caacacctat gaggtgaaga ttgacaacag ccaggtggag
     1141 tccggctcct tggaagacga ttgggacttc ctgccaccca agaagataaa ggatcctgat
     1201 gcttcaaaac cggaagactg ggatgagcgg gccaagatcg atgatcccac agactccaag
     1261 cctgaggact gggacaagcc cgagcatatc cctgaccctg atgctaagaa gcccgaggac
     1321 tgggatgaag agatggacgg agagtgggaa cccccagtga ttcagaaccc tgagtacaag
     1381 ggtgagtgga agccccggca gatcgacaac ccagattaca agggcacttg gatccaccca
     1441 gaaattgaca accccgagta ttctcccgat cccagtatct atgcctatga taactttggc
     1501 gtgctgggcc tggacctctg gcaggtcaag tctggcacca tctttgacaa cttcctcatc
     1561 accaacgatg aggcatacgc tgaggagttt ggcaacgaga cgtggggcgt aacaaaggca
     1621 gcagagaaac aaatgaagga caaacaggac gaggagcaga ggcttaagga ggaggaagaa
     1681 gacaagaaac gcaaagagga ggaggaggca gaggacaagg aggatgatga ggacaaagat
     1741 gaggatgagg aggatgagga ggacaaggag gaagatgagg aggaagatgt ccccggccag
     1801 gccaaggacg agctggcatc cgggggtgcg tcaggcggca gcggatctgg accggtcgcc
     1861 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1921 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1981 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2041 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2101 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2161 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2221 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2281 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2341 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2401 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2461 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2521 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaagcggc
     2581 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2641 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2701 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2761 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     2821 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2881 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2941 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3001 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3061 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3121 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3181 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3241 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3301 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3361 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3421 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3601 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3661 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3721 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3781 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3841 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3901 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3961 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4021 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4081 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4141 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4201 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4261 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4321 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4381 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4441 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4501 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4561 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4621 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4681 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4741 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4801 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4861 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4921 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4981 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5041 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5341 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5701 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5761 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5821 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5881 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     5941 ttagttcata gcccatatat ggagttccg
//
