LOCUS       sequence-444172-        9201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Desmosomes, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9201)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 9201)
  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-Desmoglein1-C-18
FEATURES             Location/Qualifiers
     source          1..9201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    165..212
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     237..256
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(444..1238)
                     /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             complement(444..1238)
                     /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(555..574)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1165..1184
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1273..1630)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1287..1422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1439..1630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1587..1607
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1632..1736)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1763..2218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1850..1869)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2060..2081
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2225..2346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2290..2309
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7015..7722)
                     /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     complement(7290..7309)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      7719..7728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7767..7970)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7796..7816)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7971..8274
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(8449..9037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8529..8548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg
       61 ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg
      121 ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac
      181 accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt
      241 ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc
      301 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
      361 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
      421 catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
      481 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
      541 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
      601 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
      661 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc
      721 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
      781 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
      841 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
      901 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
      961 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     1021 gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac
     1081 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     1141 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     1201 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     1261 tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     1321 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     1381 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     1441 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     1501 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     1561 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt
     1621 tccgcctcag gactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1681 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1741 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1801 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1861 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1921 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1981 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2041 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2101 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2161 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttac
     2221 gcgttaagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2281 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2341 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2401 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagttatct
     2461 agatccggtg gatccctact tgctatattg cacggtacta tactttgtga ttcgacttcg
     2521 agctgtgcta ggggaggcgg acccaatggt ttggttaaag tgatggtcag aggaactcct
     2581 catgtggcca atgctggctg tccctcccaa gctgctcagg ccaatgccac caccacttat
     2641 gccagctcca ctcagggcac cgctcccagc acccatgctg gtgccaacca ggccactgct
     2701 gcctatgcca ccgctgatcc cagtggtgcc actaattcca gttacgccag caccagaaac
     2761 aaccctctct gtcacaatga cattgtgggc attggctaac tcggggtgca tactcagact
     2821 acctatcatg ccggaagttg gttggattac tctttctgtc actaccacat ttgaagactc
     2881 tctaggatga tggatagtca gagaggtggg tagactagag cttggtgcta ttaccctttc
     2941 tgtcactata acattcgacc catctcgcaa gtcaggcatc tctaacattc catgcaaatc
     3001 agcgccagag attgggccga ccactctctc tgtcactacc acgtttgatg ctggtcggtt
     3061 atcgtgaacg tgcacagagg gcttcagagt gtcagaggtg gtgtaagact cggtcacagt
     3121 gacattacca tagcccagag gatcgagaat aggcttagga tgcagtacac caggtcccga
     3181 gggataggtg ctctcagaaa ttactgtggt agtgccgaaa tgtggggaga ttggtgggtg
     3241 tccactaaca acgggctctg tttcctgagg aaggcaaact ggttcagtgc tttgtggtgg
     3301 ccaagaagga tcaaggtctg gatatgattc ttttcctagg ctgatgtctg ccaacttctt
     3361 aaatttaggt cccagggtat ccaagaagct gtcatccagg tcttctccaa tgaagctaca
     3421 acaacccaca gagccagcag gggaacctac accttcgatg tcatatatga gcaaacagtc
     3481 attagatggg cgtccttcat cttcatctgc gtaagcatat gctttctgac agaagtagct
     3541 ttccatgaaa ttcatattca gacctccttc tctacattcc ctcatagaat ttcttcttaa
     3601 tgttccagag tattcttgac atatctcagg cattcctgaa gttttaactc cctctgttag
     3661 ttcaaatcct gtcattctct ctcctcctcc caggtcttgc atttctctgc caccatactc
     3721 atttgtataa actcctgagt tgtcaatgca ttcaattata tttgcgttat caggtggtat
     3781 ttgtggtatg acagtggtta tatccctggg ttcaggctgt ggtccttcta ctgcccatga
     3841 atgaattgct ccatctgaac attcgggaac aggctcaaag ccagctgcac tacgaggagc
     3901 acctccacaa tcacaacaga tcatcaaaaa tgggaccaat cctaagacca agaatcccat
     3961 gatgaggagt ccaatgccag caggaccaaa atgtacattg tctgataaca aatctttggc
     4021 tccgtttccg ggttcagaag aatatacttg gctagagtca gtagaagttg tgctggtatc
     4081 atagttagtg gaagaagtac tttcttgtct gccagtattg gtagtaattt tagtgttcgg
     4141 ctctgtatta gtcctgtcgt cattaccaaa actttgaatg ttaatattaa ttgtaccagt
     4201 gcaagttctt tgaagattat catctataga gagaatcgtt ccttggtatt ttcctccgag
     4261 catattgtac tgttccttgg taactttatt tttcaaagtg agtttgcctg ttcttgaatc
     4321 aacagcaagc aggtcagctg gattatttcc cattacatac ctaacagtcg ttgaaggtct
     4381 acctgtgtcc aggtcagtag ctacaaagtc tcccacttta tcatttgatc ccatattacc
     4441 agttacaaca tatgtctttg aacctggacg aaacactggg ccttcaatta catttaacac
     4501 agtcacagaa attgcagatg ctttcagttt atattgagac ataattgaat gatgaaattc
     4561 agctttattt ctgacaccaa tactgagttg cagactctgc actgagagat cccctcataa
     4621 tttccccaaa gcgtaaccat gtgtgaataa attttgagct agtagggttg cagccacgag
     4681 taagtcttcc cttgttattg tgtagccaga atgccgcaaa acttccatgc ctaagcgaac
     4741 tgttgagagt acgtttcgat ttctgactgt gttagcctgg aagtgcttgt cccaaccttg
     4801 tttctgagca tgaacgcccg caagccaaca tgttagttga agcatcaggg cgattagcag
     4861 catgatatca aaacgctctg agctgctcgt tcggctatgg cgtaggccta gtccgtaggc
     4921 aggacttttc aagtctcgga aggtttcttc aatctgcatt cgcttcgaat agatattaac
     4981 aagttgtttg ggtgttcgaa tttcaacagg taagttagtt gctagaaccc atggctcctt
     5041 tgccgacgct gagtagattt taggtgacgg gtggtgacaa tgagtccgtg tcgagcgctg
     5101 attttttcgg cctttagagc gagatttata caatagaatt tggcatgaga ttggattgct
     5161 tttagtcagc ctcttatagc ctaaagtctt tgagtgacta gatgacatat catgtaagtt
     5221 gctgataggt ttccagtttt ccgctcctag gtctgcatat tgtacttttc ctcttactcg
     5281 acttaaccag taccaaccca gcttctcaac ggatttatac catggcactt taaagccagc
     5341 atcactgaca atgagcggtg tggtgttact cggtagaatg ctcgcaaggt cggctagaaa
     5401 ttggtcatga gctttctttg aacattgctc tgaaagcggg aacgctttct cataaagagt
     5461 aacagaacga ccgtgtagtg cgactgaagc tcgcaatacc ataagtcgtt tttgctcacg
     5521 aatatcagac cagtcaacaa gtacaatggg catcgtattg cccgaacaga taaagctagc
     5581 atgccaacgg tatacagcga gtcgctcttt gtggaggtga cgattaccta acaatcggtc
     5641 gattcgtttg atgttatgtt ttgttctcgc tttggttggc aggttacggc caagttcggt
     5701 aagagtgaga gttttacagt caagtaatgc gtggcaagcc aacgttaagc tgttgagtcg
     5761 ttttaagtgt aattcggggc agaattggta aagagagtcg tgtaaaatat cgagttcgca
     5821 catcttgttg tctgattatt gatttttcgc gaaaccattt gatcatatga caagatgtgt
     5881 atccacctta acttaatgat ttttaccaaa atcattaggg gattcatcag gactctgcat
     5941 agcttcataa tctaagggct taacaacctt taaaattccc acatttgttc tttcattcat
     6001 ttctatctca aaccaatttc cttcatttcc agagataaag aaaattactg ccatccagtt
     6061 agctgagaac tcttcatcca aatcaattac tctaatttcg agcaaatttg aatttagagt
     6121 attttcttga atttctatgg tatatgaaga ctgttccatg taagggatat tatcattgac
     6181 atcgaggatt ttaatgttgc actcacattc cgctgacatg ccatctgccc caccatctcg
     6241 gtcagagcct cttacagcaa gagcatactg gccgtattgc tctctgtcta gaaaattatt
     6301 catcgttcga atttctccag tatttctgtt gataataaac attggtgaat ctgaaggttc
     6361 ttgtcttata atcttgaagg ctatttttga gttcaaattg ttcggttcat ctgcgtcagt
     6421 agcattgagt atcatcacca gtgtatttgc attagaattt tcttctattt gtcctgcaaa
     6481 tgtagccatt gaaaacactg gagggttgtc atttatatcc aaaaccctga ctctgagctc
     6541 tagaggcctc tctaaatctt ggcccattga gttcagagct cggcagtaga taatgaagaa
     6601 aggagtgacc tctcgatcaa ctatggatgt tatattaatt tcaccagttt tctgattaat
     6661 gacaaagatc ccatatggtg gctgatcaat tcctactcca gagatgcggt atgtaacttg
     6721 ctggtttgca gcacaatctg agtgaatttt ggcgattggg ttcctctttg agttgtcttc
     6781 accttcacga caggctgctg cgaacttgat ccattcacgt ttctgccttc ggattgaatg
     6841 ccatttgatg gtgccatttt tagtgttata atctcttacc tggattcgga actcactgtt
     6901 aacttctacc accaccagaa aaatgaacag cactgcaact actctgaaga aactccagtc
     6961 gcttccgctt ccgccggatg cgctgcctcc accgctgcca gatctgagtc cggacttgta
     7021 cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact gttccacgat
     7081 ggtgtagtcc tcgttgtggg acacgatgtc caacttgatg tcgacgatgt aggcgccggg
     7141 cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcggcggcgt agtggccgcc
     7201 gtccttcagc ttcagcctct tcttgatctc gctcttcagg gcgccgtcct cggggtacat
     7261 ccgctcctcg gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg
     7321 gaagttggtg ccgcgcagct tcaccttgta gatgaacacg ccgtcctgca gggaggagtc
     7381 ctggttaacg tgaataatgc cgccgtcctc gaagttcatc acgcgctccc acctgaagcc
     7441 ctcggggaag gacagcttga agtagtcggg gatgtcggct gggtgcttaa tgtagacctt
     7501 ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg ggccaccctt
     7561 ggtcaccttc agcttagcgg tctgaaaggc ctcgtagggg cggccctcgc cctcgccctc
     7621 gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaactc
     7681 cttgatgatg gccatgttat tctcctcgcc cttgctcacc atggtggcga ccggtagcgc
     7741 tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc
     7801 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     7861 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     7921 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     7981 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     8041 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     8101 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     8161 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     8221 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     8281 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actaatgcat
     8341 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8641 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8881 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     9001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     9061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     9121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9181 tatatatgag taacctgagg c
//
