LOCUS       sequence-271041-        6448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions/Membrane, Excitation: 587, Emission: 
            610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 6448)
  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-Ezrin-N-14
FEATURES             Location/Qualifiers
     source          1..6448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2489..2498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2495..3205
                     /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(2640..2658)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2908..2927
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3328..3449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3365..3384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3419..3438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3456..3911)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3593..3614)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3805..3824
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3938..4042
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4044..4401
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4044..4235
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4067..4087)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4252..4387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4436..5230
                     /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             4436..5230
                     /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(4490..4509)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5100..5119
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5418..5437)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5462..5509
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5838..6426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6327..6346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 ggtttagtga accgtcagat ccgctagcgc caccatgccg aaaccaatca atgtccgagt
      721 taccaccatg gatgcagagc tggagtttgc aatccagcca aatacaactg gaaaacagct
      781 ttttgatcag gtggtaaaga ctatcggcct ccgggaagtg tggtactttg gcctccacta
      841 tgtggataat aaaggatttc ctacctggct gaagctggat aagaaggtgt ctgcccagga
      901 ggtcaggaag gagaatcccc tccagttcaa gttccgggcc aagttctacc ctgaagatgt
      961 ggctgaggag ctcatccagg acatcaccca gaaacttttc ttcctccaag tgaaggaagg
     1021 aatccttagc gatgagatct actgcccccc tgagactgcc gtgctcttgg ggtcctacgc
     1081 tgtgcaggcc aagtttgggg actacaacaa agaagtgcac aagtctgggt acctcagctc
     1141 tgagcggctg atccctcaaa gagtgatgga ccagcacaaa cttaccaggg accagtggga
     1201 ggaccggatc caggtgtggc atgcggaaca ccgtgggatg ctcaaagata atgctatgtt
     1261 ggaatacctg aagattgctc aggacctgga aatgtatgga atcaactatt tcgagataaa
     1321 aaacaagaaa ggaacagacc tttggcttgg agttgatgcc cttggactga atatttatga
     1381 gaaagatgat aagttaaccc caaagattgg ctttccttgg agtgaaatca ggaacatctc
     1441 tttcaatgac aaaaagtttg tcattaaacc catcgacaag aaggcacctg actttgtgtt
     1501 ttatgcccca cgtctgagaa tcaacaagcg gatcctgcag ctctgcatgg gcaaccatga
     1561 gttgtatatg cgccgcagga agcctgacac catcgaggtg cagcagatga aggcccaggc
     1621 ccgggaggag aagcatcaga agcagctgga gcggcaacag ctggaaacag agaagaaaag
     1681 gagagaaacc gtggagagag agaaagagca gatgatgcgc gagaaggagg agttgatgct
     1741 gcggctgcag gactatgagg agaagacaaa gaaggcagag agagagctct cggagcagat
     1801 tcagagggcc ctgcagctgg aggaggagag gaagcgggca caggaggagg ccgagcgcct
     1861 agaggctgac cgtatggctg cactgcgggc taaggaggag ctggagagac aggcggtgga
     1921 tcagataaag agccaggagc agctggctgc ggagcttgca gaatacactg ccaagattgc
     1981 cctcctggaa gaggcgcgga ggcgcaagga ggatgaagtt gaagagtggc agcacagggc
     2041 caaagaagcc caggatgacc tggtgaagac caaggaggag ctgcacctgg tgatgacagc
     2101 acccccgccc ccaccacccc ccgtgtacga gccggtgagc taccatgtcc aggagagctt
     2161 gcaggatgag ggcgcagagc ccacgggcta cagcgcggag ctgtctagtg agggcatccg
     2221 ggatgaccgc aatgaggaga agcgcatcac tgaggcagag aagaacgagc gtgtgcagcg
     2281 gcagctgctg acgctgagca gcgagctgtc ccaggcccga gatgagaata agaggaccca
     2341 caatgacatc atccacaacg agaacatgag gcaaggccgg gacaagtaca agacgctgcg
     2401 gcagatccgg cagggcaaca ccaagcagcg catcgacgag ttcgaggccc tggcatccgg
     2461 cggtagcggc ggtggcagcg gaccggtcgc caccatggtg agcaagggcg aggaggataa
     2521 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     2581 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2641 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2701 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2761 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2821 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2881 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2941 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     3001 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     3061 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     3121 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     3181 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     3241 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3301 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3361 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3421 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3481 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3541 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3601 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3661 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3721 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3781 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3841 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3901 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3961 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4021 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4081 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4261 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4321 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4381 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4441 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4501 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4561 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4621 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4681 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4741 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4801 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4861 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4921 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4981 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5041 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5101 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5161 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5221 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5281 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5341 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5401 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5461 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5521 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5581 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5641 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5701 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5761 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5821 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5881 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5941 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6001 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6061 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6121 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6181 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6241 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6301 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6361 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6421 tggaaaaacg ccagcaacgc ggcctttt
//
