LOCUS       sequence-393319-        5851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Mitochondria, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5851)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 5851)
  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-PDHA1-N-10
FEATURES             Location/Qualifiers
     source          1..5851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             114..908
                     /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             114..908
                     /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(168..187)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     778..797
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1096..1115)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1140..1187
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1516..2104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2005..2024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2279..2582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2583..2786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2737..2757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4018..4027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4024..4734
                     /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(4169..4187)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4437..4456
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4857..4978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4894..4913)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4948..4967
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4985..5440)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5122..5143)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5334..5353
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5467..5571
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5573..5851,1..79)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5573..5764
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5596..5616)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5781..5851,1..65)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
       61 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
      121 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
      181 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
      241 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
      301 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
      361 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
      421 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
      481 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
      541 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
      601 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
      661 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
      721 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
      781 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
      841 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
      901 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
      961 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     1021 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     1081 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     1141 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     1201 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     1261 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     1321 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     1381 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1621 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     2221 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2281 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2341 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2401 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2461 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2521 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2581 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2641 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2701 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2761 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcaccgcc
     2821 accatgagga agatgctcgc cgccgtctcc cgcgtgctgt ctggcgcttc tcagaagccg
     2881 gcaagcagag tgctggtagc atcccgtaat tttgcaaatg atgctacatt tgaaattaag
     2941 aaatgtgacc ttcaccggct ggaagaaggc cctcctgtca caacagtgct caccagggag
     3001 gatgggctca aatactacag gatgatgcag actgtacgcc gaatggagtt gaaagcagat
     3061 cagctgtata aacagaaaat tattcgtggt ttctgtcact tgtgtgatgg tcaggaagct
     3121 tgctgtgtgg gcctggaggc cggcatcaac cccacagacc atctcatcac agcctaccgg
     3181 gctcacggct ttactttcac ccggggcctt tccgtccgag aaattctcgc agagcttaca
     3241 ggacgaaaag gaggttgtgc taaagggaaa ggaggatcga tgcacatgta tgccaagaac
     3301 ttctacgggg gcaatggcat cgtgggagcg caggtgcccc tgggcgctgg gattgctcta
     3361 gcctgtaagt ataatggaaa agatgaggtc tgcctgactt tatatggcga tggtgctgct
     3421 aaccagggcc agatattcga agcttacaac atggcagctt tgtggaaatt accttgtatt
     3481 ttcatctgtg agaataatcg ctatggaatg ggaacgtctg ttgagagagc ggcagccagc
     3541 actgattact acaagagagg cgatttcatt cctgggctga gagtggatgg aatggatatc
     3601 ctgtgcgtcc gagaggcaac aaggtttgct gctgcctatt gtagatctgg gaaggggccc
     3661 atcctgatgg agctgcagac ttaccgttac cacggacaca gtatgagtga ccctggagtc
     3721 agttaccgta cacgagaaga aattcaggaa gtaagaagta agagtgaccc tattatgctt
     3781 ctcaaggaca ggatggtgaa cagcaatctt gccagtgtgg aagaactaaa ggaaattgat
     3841 gtggaagtga ggaaggagat tgaggatgct gcccagtttg ccacggccga tcctgagcca
     3901 cctttggaag agctgggcta ccacatctac tccagcgacc caccttttga agttcgtggt
     3961 gccaatcagt ggatcaagtt taagtcagtc agtagctccg gaggggatcc accggtcgcc
     4021 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     4081 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     4141 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     4201 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     4261 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     4321 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4381 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4441 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4501 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4561 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4621 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4681 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaagcggc
     4741 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4801 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4861 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4921 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4981 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5041 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5101 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5161 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5221 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5281 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5341 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5401 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     5461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     5581 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5761 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5821 cccatggctg actaattttt tttatttatg c
//
