LOCUS       sequence-241977-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lamin A mutant (R527P), C-terminally tagged with mRFP. Mutation 
            associated with EDMD myopathy..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6662)
  TITLE     Schirmer lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6662)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Lamin A (R527P)-mRFP
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    624..680
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2657..3331
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVIQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2787..2805)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3249..3272
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3465..3586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3556..3575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3593..4048)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3730..3751)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3942..3961
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4075..4179
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4181..4538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4181..4372
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4204..4224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4389..4524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4573..5367
                     /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             4573..5367
                     /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(4627..4646)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5237..5256
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5555..5574)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5599..5646
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5975..6563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6464..6483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      661 gtaccgcggg cccgggatcc gaatggccat ggagaccccg tcccagcggc gcgccacccg
      721 cagcggggcg caggccagct ccactccgct gtcgcccacc cgcatcaccc ggctgcagga
      781 gaaggaggac ctgcaggagc tcaatgatcg cttggcggtc tacatcgacc gtgtgcgctc
      841 gctggaaacg gagaacgcag ggctgcgcct tcgcatcacc gagtctgaag aggtggtcag
      901 ccgcgaggtg tccggcatca aggccgccta cgaggccgag ctcggggatg cccgcaagac
      961 ccttgactca gtagccaagg agcgcgcccg cctgcagctg gagctgagca aagtgcgtga
     1021 ggagtttaag gagctgaaag cgcgcaatac caagaaggag ggtgacctga tagctgctca
     1081 ggctcggctg aaggacctgg aggctctgct gaactccaag gaggccgcac tgagcactgc
     1141 tctcagtgag aagcgcacgc tggagggcga gctgcatgat ctgcggggcc aggtggccaa
     1201 gcttgaggca gccctaggtg aggccaagaa gcaacttcag gatgagatgc tgcggcgggt
     1261 ggatgctgag aacaggctgc agaccatgaa ggaggaactg gacttccaga agaacatcta
     1321 cagtgaggag ctgcgtgaga ccaagcgccg tcatgagacc cgactggtgg agattgacaa
     1381 tgggaagcag cgtgagtttg agagccggct ggcggatgcg ctgcaggaac tgcgggccca
     1441 gcatgaggac caggtggagc agtataagaa ggagctggag aagacttatt ctgccaagct
     1501 ggacaatgcc aggcagtctg ctgagaggaa cagcaacctg gtgggggctg cccacgagga
     1561 gctgcagcag tcgcgcatcc gcatcgacag cctctctgcc cagctcagcc agctccagaa
     1621 gcagctggca gccaaggagg cgaagcttcg agacctggag gactcactgg cccgtgagcg
     1681 ggacaccagc cggcggctgc tggcggaaaa ggagcgggag atggccgaga tgcgggcaag
     1741 gatgcagcag cagctggacg agtaccagga gcttctggac atcaagctgg ccctggacat
     1801 ggagatccac gcctaccgca agctcttgga gggcgaggag gagaggctac gcctgtcccc
     1861 cagccctacc tcgcagcgca gccgtggccg tgcttcctct cactcatccc agacacaggg
     1921 tgggggcagc gtcaccaaaa agcgcaaact ggagtccact gagagccgca gcagcttctc
     1981 acagcacgca cgcactagcg ggcgcgtggc cgtggaggag gtggatgagg agggcaagtt
     2041 tgtccggctg cgcaacaagt ccaatgagga ccagtccatg ggcaattggc agatcaagcg
     2101 ccagaatgga gatgatccct tgctgactta ccggttccca ccaaagttca ccctgaaggc
     2161 tgggcaggtg gtgacgatct gggctgcagg agctggggcc acccacagcc cccctaccga
     2221 cctggtgtgg aaggcacaga acacctgggg ctgcgggaac agcctgccta cggctctcat
     2281 caactccact ggggaagaag tggccatgcg caagctggtg cgctcagtga ctgtggttga
     2341 ggacgacgag gatgaggatg gagatgacct gctccatcac caccacggct cccactgcag
     2401 cagctcgggg gaccccgctg agtacaacct gcgctcgcgc accgtgctgt gcgggacctg
     2461 cgggcagcct gccgacaagg catctgccag cggctcagga gcccaggtgg gcggacccat
     2521 ctcctctggc tcttctgcct ccagtgtcac ggtcactcgc agctaccgca gtgtgggggg
     2581 cagtgggggt ggcagcttcg gggacaatct ggtcacccgc tcctacggtg gcggtggctc
     2641 tggaggtggt gggtccatgg cctcctccga ggacgtcatc aaggagttca tgcgcttcaa
     2701 ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2761 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     2821 cttcgcctgg gacatcctgt cccctcagtt ccagtacggc tccaaggcct acgtgaagca
     2881 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2941 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     3001 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     3061 aattcagaag aagacgatgg gctgggaggc ctccaccgag cggatgtacc ccgaggacgg
     3121 cgccctgaag ggcgagatca agatgaggct gaagctgaag gacggcggcc actacgacgc
     3181 cgaggtcaag accacctaca tggccaagaa gcccgtgcag ctgcccggcg cctacaagac
     3241 cgacatcaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3301 gcgcgccgag ggccgccact ccaccggcgc cctgtacaag taagcggccg cgactctaga
     3361 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3421 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3481 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3541 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3601 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3661 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3721 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3781 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3841 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3901 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3961 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4021 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4081 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4141 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4201 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4261 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4381 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4441 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4501 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4561 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4621 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4681 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4741 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4801 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4861 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4921 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4981 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5041 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5101 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5161 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5221 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5281 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5341 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5401 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5461 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5521 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5581 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5641 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5701 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5761 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5821 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5881 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6121 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6601 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6661 gt
//
