LOCUS       sequence-127244-        6051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Centrosomes, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6051)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 6051)
  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-Gamma-Tubulin-17
FEATURES             Location/Qualifiers
     source          1..6051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..129
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(428..447)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    472..519
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      848..1436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1337..1356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1611..1914
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1915..2118
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2069..2089
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3551..3560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3557..4267
                     /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     3970..3989
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4389..4510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4426..4445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4480..4499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4517..4972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4654..4675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4866..4885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5105..5462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5105..5296
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5128..5148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5313..5448
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5497..6051,1..240)
                     /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             join(5497..6051,1..240)
                     /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(5551..5570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
       61 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
      121 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
      181 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
      241 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
      301 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      361 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
      421 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
      481 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
      541 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
      601 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
      661 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
      721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      781 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      961 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     1561 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1621 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1681 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1741 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1801 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1861 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1921 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1981 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2041 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2101 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca ccatgccgag
     2161 ggaaatcatc accctacagt tgggccagtg cggcaatcag attgggttcg agttctggaa
     2221 acagctgtgc gccgagcatg gtatcagccc cgagggcatc gtggaggagt tcgccaccga
     2281 gggcactgac cacaaggacg tctttttcta ccaggcagac gatgagcact acatcccccg
     2341 ggccgtgctg ctggacttgg aaccccgggt gatccactcc atcctcaact ccccctatgc
     2401 caagctctac aacccagaga acatctacct gtcggaacat ggaggaggag ctggcaacaa
     2461 ctgggccagc ggattctccc agggagaaaa gatccatgag gacatttttg acatcataga
     2521 ccgggaggca gatggtagtg acagtctaga gggctttgtg ctgtgtcact ccattgctgg
     2581 ggggacaggc tctggactgg gttcctacct cttagaacgg ctgaatgaca ggtatcctaa
     2641 gaagctggtg cagacatact cagtgtttcc caaccaggac gagatgagcg atgtggtggt
     2701 ccagccttac aattcactcc tcacactcaa gaggctgacg cagaatgcag actgtgtggt
     2761 ggtgctggac aacacagccc tgaaccggat tgccacagac cgcctgcaca tccagaaccc
     2821 atccttctcc cagatcaacc agctggtgtc taccatcatg tcagccagca ccaccaccct
     2881 gcgctaccct ggctacatga acaatgacct catcggcctc atcgcctcgc tcattcccac
     2941 cccacggctc cacttcctca tgaccggcta cacccctctc actacggacc agtcagtggc
     3001 cagcgtgagg aagaccacgg tcctggatgt catgaggcgg ctgctgcagc ccaagaacgt
     3061 gatggtgtcc acaggccgag accgccagac caaccactgc tacatcgcca tcctcaacat
     3121 catccaggga gaggtggacc ccacccaggt ccacaagagc ttgcagagga tccgggaacg
     3181 caagttggcc aacttcatcc cgtggggccc cgccagcatc caggtggccc tgtcgaggaa
     3241 gtctccctac ctgccctcgg cccaccgggt cagcgggctc atgatggcca accacaccag
     3301 catctcctcg ctcttcgaga gaacctgtcg ccagtatgac aagctgcgta agcgggaggc
     3361 cttcctggag cagttccgca aggaggacat gttcaaggac aactttgatg agatggacac
     3421 atccagggag attgtgcagc agctcatcga tgagtaccat gcggccacac ggccagacta
     3481 catctcctgg ggcactcagg agcagcgaat tctgcagtcg acggtaccgc gggcccggga
     3541 tccaccggtc gccaccatgg tgagcaaggg cgaggagaat aacatggcca tcatcaagga
     3601 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3661 gggcgagggc gagggccgcc cctacgaggc ctttcagacc gctaagctga aggtgaccaa
     3721 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3781 ggtctacatt aagcacccag ccgacatccc cgactacttc aagctgtcct tccccgaggg
     3841 cttcaggtgg gagcgcgtga tgaacttcga ggacggcggc attattcacg ttaaccagga
     3901 ctcctccctg caggacggcg tgttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3961 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctccg aggagcggat
     4021 gtaccccgag gacggcgccc tgaagagcga gatcaagaag aggctgaagc tgaaggacgg
     4081 cggccactac gccgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     4141 cggcgcctac atcgtcgaca tcaagttgga catcgtgtcc cacaacgagg actacaccat
     4201 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     4261 caagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     4321 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4381 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4441 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4501 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4561 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4621 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4681 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4741 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4801 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4861 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4921 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5101 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5161 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5221 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5281 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5341 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5401 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5461 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5521 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5581 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5641 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     5701 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5761 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5821 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5881 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5941 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6001 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt c
//
