LOCUS       sequence-174354-        6703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytoskeleton, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 6703)
  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-Palladin-C-7
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..708
                     /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     414..433
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2879..3000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2916..2935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2970..2989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3007..3462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3144..3165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3356..3375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3489..3593
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3595..3952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3595..3786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3618..3638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3803..3938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3987..4781
                     /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             3987..4781
                     /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(4041..4060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4651..4670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4969..4988)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5013..5060
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5389..5977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5878..5897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6152..6455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6456..6659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6610..6630
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6698..6703,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gaataacatg gccatcatca aggagttcat gcgcttcaag
       61 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      121 cgcccctacg aggcctttca gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc
      181 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggtcta cattaagcac
      241 ccagccgaca tccccgacta cttcaagctg tccttccccg agggcttcag gtgggagcgc
      301 gtgatgaact tcgaggacgg cggcattatt cacgttaacc aggactcctc cctgcaggac
      361 ggcgtgttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
      421 atgcagaaga agaccatggg ctgggaggcc tccgaggagc ggatgtaccc cgaggacggc
      481 gccctgaaga gcgagatcaa gaagaggctg aagctgaagg acggcggcca ctacgccgcc
      541 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
      601 gacatcaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga acagtacgaa
      661 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
      721 tctcgagtga gcgctctggc ctcccgcgct accgccatgc agtcctccgg ctccttcaac
      781 tacgcgcgcc ctaagcagtt catcgcggcg cagaacctgg gtcccgcatc ggggctgccc
      841 acgccaacct ccagccccag ctcctccagc ttgccgtcac cattgtcccc cacgcccagg
      901 cccttcggcc gcgcgcccgg gccgcccttc gtggaacccg aggccatgtg gggcccgtcc
      961 tcgccctcgc cgccaccgcc gccacctccg gtcttcagcc cctctgcagc ctacccggtg
     1021 cccgacgtgt tcccactgcc accgccgccg ccaccgctac ctagctccac ctcacactgc
     1081 gcctcgcccg cccgcttcgg ccccagccag acgcccgcag ccttcctcag cgccctgctg
     1141 ccctctcagc caccgcctgt cgctgtcaac gccctggggc tgcccaaggg agtcaccccc
     1201 gcggggttcc caaagaagtc cagtagaact gctagaattg cctctgatga ggagattcaa
     1261 ggcacaaagg atgctgtcat ccaagacctg gaacggaagc ttcgcttcaa ggaggacctt
     1321 ctgaacaatg gccaaccgag gctaacctat gaggaaagaa tggctcgccg cctgcttgga
     1381 gccgacagcg caaacgtctt caacatccag gagccagagg aaacggcagc caatcaggag
     1441 tacaaagtct ctagctgcga gcagaggctg attagcgaga ttgaatacag gctggagcgc
     1501 tcccctgtgg atgagtcggg agacgaggtg caggatccag atgtgcctgt ggagaacgca
     1561 acagctccct tctttgagat gaagctgaaa cactacaaga tctttgaggg gatgccggtg
     1621 actttcacgt gtcgagtggc tgggaatcca aagccaaaga tctattggtt taaagatggg
     1681 aagcagattt ctccgaagag cgatcactac accattcaga gagaccttga tgggacctgc
     1741 tctctccaca ccacggcctc taccctagac gacgatggga actacaccat catggctgcc
     1801 aaccctcagg gtcgcgtcag ttgtacagga aggctaatgg tacaggctgt caaccaaaga
     1861 ggccgcagtc cccgctctcc ctcaggccat cctcatgcca gaaggcctcg ctctcgatca
     1921 cgggacagtg gagatgaaaa cgagcccatt caggagcgat tcttcagacc tcacttcctg
     1981 caggctcctg gagacctgac cgttcaggaa ggcaagctct gcaggatgga ctgcaaggtc
     2041 agtgggttac caaccccaga tctcagctgg caactagacg gaaagcccat acgccccgac
     2101 agtgctcaca agatgctggt ccgtgagaat ggggtccact ccctcattat agagccagtc
     2161 acgtcccggg acgccggcat ctacacatgt attgccacca acagagcagg acagaactcg
     2221 tttaacctgg agcttgtggt tgctgctaag gaagcacaca aggcccctgt gtttatggag
     2281 aagctacaga acacgggggt tgctgatgga tacccagtgc ggctagaatg ccgtgtctcc
     2341 ggagtgccgc cacctcagat attttggaag aaagaaaatg aatcgctcac tcatagcact
     2401 gagcgagtaa gcatgcacca ggataatcat ggctacatct gcctgctcat ccagggagcc
     2461 acaaaggaag acgctgggtg gtacactgtg tctgccaaga acgaagcagg cattgtgtcc
     2521 tgcactgccc ggctggatgt ctacacccag tggcaccagc agccacagac caccaagcca
     2581 aaaaaagtac ggccctcggc cagtcgctac gcagcacttt cggaccaggg actagacatc
     2641 aaagccgctt tccaacctga agccagccca tctcacctga ctctgaacag cggcttggta
     2701 gaaagtgaag acctgtgaga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
     2761 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2821 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3001 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3061 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3481 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3541 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3601 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3781 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3901 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3961 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4021 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4081 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4141 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4201 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4261 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4321 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4381 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4441 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4501 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4561 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4621 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4681 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4741 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4801 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4861 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4921 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4981 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5041 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5101 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5161 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5221 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5281 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6001 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6061 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     6121 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     6241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     6361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     6421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     6481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     6541 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     6601 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     6661 gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc acc
//
