LOCUS       sequence-152016-        7417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Alpha Actinin, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7417)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 7417)
  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-Alpha-Actinin-19
FEATURES             Location/Qualifiers
     source          1..7417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3371..3380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3377..4087
                     /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(3522..3540)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3790..3809
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4212..4333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4249..4268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4303..4322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4340..4795)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4477..4498)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4689..4708
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4822..4926
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4928..5285
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4928..5119
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4951..4971)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5136..5271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5320..6114
                     /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             5320..6114
                     /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(5374..5393)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5984..6003
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6302..6321)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6346..6393
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6722..7310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7211..7230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgcacc atcatggacc attatgattc
      661 tcagcaaacc aacgattaca tgcagccaga agaggactgg gaccgggacc tgctcctgga
      721 cccggcctgg gagaagcagc agagaaagac attcacggca tggtgtaact cccacctccg
      781 gaaggcgggg acacagatcg agaacatcga agaggacttc cgggatggcc tgaagctcat
      841 gctgctgctg gaggtcatct caggtgaacg cttggccaag ccagagcgag gcaagatgag
      901 agtgcacaag atctccaacg tcaacaaggc cctggatttc atagccagca aaggcgtcaa
      961 actggtgtcc atcggagccg aagaaatcgt ggatgggaat gtgaagatga ccctgggcat
     1021 gatctggacc atcatcctgc gctttgccat ccaggacatc tccgtggaag agacttcagc
     1081 caaggaaggg ctgctcctgt ggtgtcagag aaagacagcc ccttacaaaa atgtcaacat
     1141 ccagaacttc cacataagct ggaaggatgg cctcggcttc tgtgctttga tccaccgaca
     1201 ccggcccgag ctgattgact acgggaagct gcggaaggat gatccactca caaatctgaa
     1261 tacggctttt gacgtggcag agaagtacct ggacatcccc aagatgctgg atgccgaaga
     1321 catcgttgga actgcccgac cggatgagaa agccatcatg acttacgtgt ctagcttcta
     1381 ccacgccttc tctggagccc agaaggcgga gacagcagcc aatcgcatct gcaaggtgtt
     1441 ggccgtcaac caggagaacg agcagcttat ggaagactac gagaagctgg ccagtgatct
     1501 gttggagtgg atccgccgca caatcccgtg gctggagaac cgggtgcccg agaacaccat
     1561 gcatgccatg caacagaagc tggaggactt ccgggactac cggcgcctgc acaagccgcc
     1621 caaggtgcag gagaagtgcc agctggagat caacttcaac acgctgcaga ccaagctgcg
     1681 gctcagcaac cggcctgcct tcatgccctc tgagggcagg atggtctcgg acatcaacaa
     1741 tgcctggggc tgcctggagc aggtggagaa gggctatgag gagtggttgc tgaatgagat
     1801 ccggaggctg gagcgactgg accacctggc agagaagttc cggcagaagg cctccatcca
     1861 cgaggcctgg actgacggca aagaggccat gctgcgacag aaggactatg agaccgccac
     1921 cctctcggag atcaaggcct tgctcaagaa gcatgaggcc ttcgagagtg acctggctgc
     1981 ccaccaggac cgtgtggagc agattgccgc catcgcacag gagctcaatg agctggacta
     2041 ttatgactca cccagtgtca acgcccgttg ccaaaagatc tgtgaccagt gggacaatct
     2101 gggggcccta actcagaagc gaagggaagc tctggagcgg accgagaaac tgctggagac
     2161 cattgaccag ctgtacttgg agtatgccaa gcgggctgca cccttcaaca actggatgga
     2221 gggggccatg gaggacctgc aggacacctt cattgtgcac accattgagg agatccaggg
     2281 actgaccaca gcccatgagc agttcaaggc caccctccct gatgccgaca aggagcgcct
     2341 ggccatcctg ggcatccaca atgaggtgtc caagattgtc cagacctacc acgtcaatat
     2401 ggcgggcacc aacccctaca caaccatcac gcctcaggag atcaatggca aatgggacca
     2461 cgtgcggcag ctggtgcctc ggagggacca agctctgacg gaggagcatg cccgacagca
     2521 gcacaatgag aggctacgca agcagtttgg agcccaggcc aatgtcatcg ggccctggat
     2581 ccagaccaag atggaggaga tcgggaggat ctccattgag atgcatggga ccctggagga
     2641 ccagctcagc cacctgcggc agtatgagaa gagcatcgtc aactacaagc caaagattga
     2701 tcagctggag ggcgaccacc agctcatcca ggaggcgctc atcttcgaca acaagcacac
     2761 caactacacc atggagcaca tccgtgtggg ctgggagcag ctgctcacca ccatcgccag
     2821 gaccatcaat gaggtagaga accagatcct gacccgggat gccaagggca tcagccagga
     2881 gcagatgaat gagttccggg cctccttcaa ccactttgac cgggatcact ccggcacact
     2941 gggtcccgag gagttcaaag cctgcctcat cagcttgggt tatgatattg gcaacgaccc
     3001 ccagggagaa gcagaatttg cccgcatcat gagcattgtg gaccccaacc gcctgggggt
     3061 agtgacattc caggccttca ttgacttcat gtcccgcgag acagccgaca cagatacagc
     3121 agaccaagtc atggcttcct tcaagatcct ggctggggac aagaactaca ttaccatgga
     3181 cgagctgcgc cgcgagctgc cacccgacca ggctgagtac tgcatcgcgc ggatggcccc
     3241 ctacaccggc cccgactccg tgccaggtgc tctggactac atgtccttct ccacggcgct
     3301 gtacggcgag agtgacctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
     3361 tccaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     3421 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3481 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3541 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3601 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     3661 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3721 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3781 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3841 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3901 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3961 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     4021 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     4081 caagtaaagc gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     4141 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4201 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4261 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4321 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4381 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4441 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4501 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4561 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4621 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4681 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4741 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4861 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4921 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4981 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5041 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5101 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5161 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5221 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5281 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5341 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5401 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5461 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     5521 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5581 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5641 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5701 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5761 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5821 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5881 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5941 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6001 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6061 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6121 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6181 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6241 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     6301 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     6361 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     6421 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6481 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6541 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6601 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6661 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6721 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6781 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6841 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6901 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6961 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7021 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7081 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7141 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7201 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7261 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7321 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7381 cgttatcccc tgattctgtg gataaccgta ttaccgc
//
