LOCUS       sequence-241969-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lamin A mutant (R298C), C-terminally tagged with Mrfp. Mutation 
            associated with CMT neuropathy..
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  JOURNAL   Unpublished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            "Unpublished"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 (R298C)-mRFP
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        121..424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        425..628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    669..725
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2702..3376
                     /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(2832..2850)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3294..3317
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3510..3631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3547..3566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3601..3620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3638..4093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3775..3796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3987..4006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4120..4224
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4226..4583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4226..4417
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4249..4269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4434..4569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4618..5412
                     /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             4618..5412
                     /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(4672..4691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5282..5301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5600..5619)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5644..5691
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6020..6608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6509..6528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
       61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      601 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      661 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      721 gatccgaatg gccatggaga ccccgtccca gcggcgcgcc acccgcagcg gggcgcaggc
      781 cagctccact ccgctgtcgc ccacccgcat cacccggctg caggagaagg aggacctgca
      841 ggagctcaat gatcgcttgg cggtctacat cgaccgtgtg cgctcgctgg aaacggagaa
      901 cgcagggctg cgccttcgca tcaccgagtc tgaagaggtg gtcagccgcg aggtgtccgg
      961 catcaaggcc gcctacgagg ccgagctcgg ggatgcccgc aagacccttg actcagtagc
     1021 caaggagcgc gcccgcctgc agctggagct gagcaaagtg cgtgaggagt ttaaggagct
     1081 gaaagcgcgc aataccaaga aggagggtga cctgatagct gctcaggctc ggctgaagga
     1141 cctggaggct ctgctgaact ccaaggaggc cgcactgagc actgctctca gtgagaagcg
     1201 cacgctggag ggcgagctgc atgatctgcg gggccaggtg gccaagcttg aggcagccct
     1261 aggtgaggcc aagaagcaac ttcaggatga gatgctgcgg cgggtggatg ctgagaacag
     1321 gctgcagacc atgaaggagg aactggactt ccagaagaac atctacagtg aggagctgcg
     1381 tgagaccaag cgccgtcatg agacccgact ggtggagatt gacaatggga agcagcgtga
     1441 gtttgagagc cggctggcgg atgcgctgca ggaactgcgg gcccagcatg aggaccaggt
     1501 ggagcagtat aagaaggagc tggagaagac ttattctgcc aagctggaca atgccaggca
     1561 gtctgctgag aggaacagca acctggtggg ggctgcccac gaggagctgc agcagtcgcg
     1621 catctgcatc gacagcctct ctgcccagct cagccagctc cagaagcagc tggcagccaa
     1681 ggaggcgaag cttcgagacc tggaggactc actggcccgt gagcgggaca ccagccggcg
     1741 gctgctggcg gaaaaggagc gggagatggc cgagatgcgg gcaaggatgc agcagcagct
     1801 ggacgagtac caggagcttc tggacatcaa gctggccctg gacatggaga tccacgccta
     1861 ccgcaagctc ttggagggcg aggaggagag gctacgcctg tcccccagcc ctacctcgca
     1921 gcgcagccgt ggccgtgctt cctctcactc atcccagaca cagggtgggg gcagcgtcac
     1981 caaaaagcgc aaactggagt ccactgagag ccgcagcagc ttctcacagc acgcacgcac
     2041 tagcgggcgc gtggccgtgg aggaggtgga tgaggagggc aagtttgtcc ggctgcgcaa
     2101 caagtccaat gaggaccagt ccatgggcaa ttggcagatc aagcgccaga atggagatga
     2161 tcccttgctg acttaccggt tcccaccaaa gttcaccctg aaggctgggc aggtggtgac
     2221 gatctgggct gcaggagctg gggccaccca cagcccccct accgacctgg tgtggaaggc
     2281 acagaacacc tggggctgcg ggaacagcct gcgtacggct ctcatcaact ccactgggga
     2341 agaagtggcc atgcgcaagc tggtgcgctc agtgactgtg gttgaggacg acgaggatga
     2401 ggatggagat gacctgctcc atcaccacca cggctcccac tgcagcagct cgggggaccc
     2461 cgctgagtac aacctgcgct cgcgcaccgt gctgtgcggg acctgcgggc agcctgccga
     2521 caaggcatct gccagcggct caggagccca ggtgggcgga cccatctcct ctggctcttc
     2581 tgcctccagt gtcacggtca ctcgcagcta ccgcagtgtg gggggcagtg ggggtggcag
     2641 cttcggggac aatctggtca cccgctccta cggtggcggt ggctctggag gtggtgggtc
     2701 catggcctcc tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg
     2761 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2821 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     2881 cctgtcccct cagttccagt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     2941 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3001 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3061 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaattc agaagaagac
     3121 gatgggctgg gaggcctcca ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3181 gatcaagatg aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac
     3241 ctacatggcc aagaagcccg tgcagctgcc cggcgcctac aagaccgaca tcaagctgga
     3301 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg
     3361 ccactccacc ggcgccctgt acaagtaagc ggccgcgact ctagatcata atcagccata
     3421 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3481 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3601 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     3661 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3721 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3781 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3841 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3901 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3961 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4021 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4081 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4141 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4201 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4261 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4441 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4501 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4561 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4621 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4681 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4741 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4801 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4861 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4921 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4981 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5041 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5101 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     5161 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5221 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5281 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5341 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5401 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5461 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5521 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5581 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5641 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5701 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5761 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5821 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5881 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5941 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6661 ct
//
