LOCUS       sequence-198676-        5565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: MT End Binding Protein, Excitation: 568, Emission: 
            592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5565)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 5565)
  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: mApple-EB3-C-20
FEATURES             Location/Qualifiers
     source          1..5565
                     /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      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1260
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     966..985
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2293..2414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2330..2349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2384..2403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2421..2876)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2558..2579)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2770..2789
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2903..3007
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3009..3366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3009..3200
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3032..3052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3217..3352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3401..4195
                     /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             3401..4195
                     /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(3455..3474)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4065..4084
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4383..4402)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4427..4474
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4803..5391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5292..5311
                     /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 ccggtcgcca ccatggtgag caagggcgag gagaataaca tggccatcat caaggagttc
      601 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      661 gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt
      721 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggtc
      781 tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc
      841 aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta ttcacgttaa ccaggactcc
      901 tccctgcagg acggcgtgtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      961 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccgagga gcggatgtac
     1021 cccgaggacg gcgccctgaa gagcgagatc aagaagaggc tgaagctgaa ggacggcggc
     1081 cactacgccg ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1141 gcctacatcg tcgacatcaa gttggacatc gtgtcccaca acgaggacta caccatcgtg
     1201 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1261 tccggactca gatctggcag cggtggaggc agcgcatccg gcggaagcgg aagcggggga
     1321 gccgtcaatg tgtactccac atctgtgacc agtgaaaatc tgagtcgcca tgatatgctt
     1381 gcatgggtca acgactccct gcacctcaac tataccaaga tagaacagct ttgttcaggg
     1441 gcagcctact gccagttcat ggacatgctc ttccccggct gtgtgcactt gaggaaagtg
     1501 aagttccagg ccaaactaga gcatgaatac atccacaact tcaaggtgct gcaagcagct
     1561 ttcaagaaga tgggtgttga caaaatcatt cctgtagaga aattagtgaa aggaaaattc
     1621 caagataatt ttgagtttat tcagtggttt aagaaattct ttgacgcaaa ctatgatgga
     1681 aaggattaca accctctgct ggcgcggcag ggccaggacg tagcgccacc tcctaaccca
     1741 ggtgatcaga tattcaacaa atccaagaaa ctcattggca cagcagttcc acagaggacg
     1801 tcccccacag gcccaaaaaa catgcagacc tctggccggc tgagcaatgt ggcccccccc
     1861 tgcattctcc ggaagaatcc tccatcagcc cgaaatggcg gccatgagac tgatgcccaa
     1921 attcttgaac tcaaccaaca gctggtggac ttgaagctga cagtggatgg gctggagaag
     1981 gaacgtgact tctacttcag caaacttcgt gacatcgagc tcatctgcca ggagcatgaa
     2041 agtgaaaaca gccctgttat ctcaggcatc attggcatcc tctatgccac agaggaagga
     2101 ttcgcacccc ctgaggacga tgagattgaa gagcatcaac aagaagacca ggacgagtac
     2161 taaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     2221 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2281 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2341 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2401 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2461 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2521 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2581 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2641 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2701 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2761 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2821 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     2881 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2941 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3001 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3061 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3121 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3181 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3361 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3421 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3481 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3541 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3601 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3661 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3721 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3781 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3841 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3901 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3961 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4021 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4081 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4141 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4201 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4261 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4321 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4381 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4441 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4501 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4561 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4621 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4681 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4741 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4801 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4861 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4921 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4981 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5041 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5101 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5161 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5221 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5281 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5341 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5401 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5461 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     5521 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
