LOCUS       sequence-251113-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lamin A mutant (R377H), 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  JOURNAL   Unpublished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            "Unpublished"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 (R377H)-mRFP
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..346
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        347..550
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    591..647
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2624..3298
                     /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(2754..2772)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3216..3239
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3432..3553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3469..3488)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3523..3542
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3560..4015)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3697..3718)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3909..3928
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4042..4146
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4148..4505
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4148..4339
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4171..4191)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4356..4491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4540..5334
                     /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             4540..5334
                     /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(4594..4613)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5204..5223
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5522..5541)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5566..5613
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5942..6530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6431..6450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
       61 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      121 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      181 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      241 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      301 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      361 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      421 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      481 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      541 aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag
      601 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccgaa tggccatgga
      661 gaccccgtcc cagcggcgcg ccacccgcag cggggcgcag gccagctcca ctccgctgtc
      721 gcccacccgc atcacccggc tgcaggagaa ggaggacctg caggagctca atgatcgctt
      781 ggcggtctac atcgaccgtg tgcgctcgct ggaaacggag aacgcagggc tgcgccttcg
      841 catcaccgag tctgaagagg tggtcagccg cgaggtgtcc ggcatcaagg ccgcctacga
      901 ggccgagctc ggggatgccc gcaagaccct tgactcagta gccaaggagc gcgcccgcct
      961 gcagctggag ctgagcaaag tgcgtgagga gtttaaggag ctgaaagcgc gcaataccaa
     1021 gaaggagggt gacctgatag ctgctcaggc tcggctgaag gacctggagg ctctgctgaa
     1081 ctccaaggag gccgcactga gcactgctct cagtgagaag cgcacgctgg agggcgagct
     1141 gcatgatctg cggggccagg tggccaagct tgaggcagcc ctaggtgagg ccaagaagca
     1201 acttcaggat gagatgctgc ggcgggtgga tgctgagaac aggctgcaga ccatgaagga
     1261 ggaactggac ttccagaaga acatctacag tgaggagctg cgtgagacca agcgccgtca
     1321 tgagacccga ctggtggaga ttgacaatgg gaagcagcgt gagtttgaga gccggctggc
     1381 ggatgcgctg caggaactgc gggcccagca tgaggaccag gtggagcagt ataagaagga
     1441 gctggagaag acttattctg ccaagctgga caatgccagg cagtctgctg agaggaacag
     1501 caacctggtg ggggctgccc acgaggagct gcagcagtcg cgcatccgca tcgacagcct
     1561 ctctgcccag ctcagccagc tccagaagca gctggcagcc aaggaggcga agcttcgaga
     1621 cctggaggac tcactggccc gtgagcggga caccagccgg cggctgctgg cggaaaagga
     1681 gcgggagatg gccgagatgc gggcaaggat gcagcagcag ctggacgagt accaggagct
     1741 tctggacatc aagctggccc tggacatgga gatccacgcc taccacaagc tcttggaggg
     1801 cgaggaggag aggctacgcc tgtcccccag ccctacctcg cagcgcagcc gtggccgtgc
     1861 ttcctctcac tcatcccaga cacagggtgg gggcagcgtc accaaaaagc gcaaactgga
     1921 gtccactgag agccgcagca gcttctcaca gcacgcacgc actagcgggc gcgtggccgt
     1981 ggaggaggtg gatgaggagg gcaagtttgt ccggctgcgc aacaagtcca atgaggacca
     2041 gtccatgggc aattggcaga tcaagcgcca gaatggagat gatcccttgc tgacttaccg
     2101 gttcccacca aagttcaccc tgaaggctgg gcaggtggtg acgatctggg ctgcaggagc
     2161 tggggccacc cacagccccc ctaccgacct ggtgtggaag gcacagaaca cctggggctg
     2221 cgggaacagc ctgcgtacgg ctctcatcaa ctccactggg gaagaagtgg ccatgcgcaa
     2281 gctggtgcgc tcagtgactg tggttgagga cgacgaggat gaggatggag atgacctgct
     2341 ccatcaccac cacggctccc actgcagcag ctcgggggac cccgctgagt acaacctgcg
     2401 ctcgcgcacc gtgctgtgcg ggacctgcgg gcagcctgcc gacaaggcat ctgccagcgg
     2461 ctcaggagcc caggtgggcg gacccatctc ctctggctct tctgcctcca gtgtcacggt
     2521 cactcgcagc taccgcagtg tggggggcag tgggggtggc agcttcgggg acaatctggt
     2581 cacccgctcc tacggtggcg gtggctctgg aggtggtggg tccatggcct cctccgagga
     2641 cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     2701 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2761 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcca
     2821 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2881 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2941 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     3001 caccaacttc ccctccgacg gccccgtaat tcagaagaag acgatgggct gggaggcctc
     3061 caccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaaga tgaggctgaa
     3121 gctgaaggac ggcggccact acgacgccga ggtcaagacc acctacatgg ccaagaagcc
     3181 cgtgcagctg cccggcgcct acaagaccga catcaagctg gacatcacct cccacaacga
     3241 ggactacacc atcgtggaac agtacgagcg cgccgagggc cgccactcca ccggcgccct
     3301 gtacaagtaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     3361 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3421 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3481 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3541 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3601 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3661 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3721 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3781 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3841 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3901 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3961 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4021 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4081 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4141 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4201 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4321 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4381 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4441 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4501 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4561 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4621 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4681 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4741 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4801 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4861 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4921 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4981 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5041 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5101 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5161 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5221 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5281 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5341 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5401 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5461 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5521 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5581 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5641 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5701 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5761 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5821 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6061 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6541 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6601 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     6661 tc
//
