LOCUS       sequence-241964-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lamin A mutant (R482Q), C-terminally tagged with mRFP. Mutation 
            associated with FPLD lipodystrophy..
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  JOURNAL   Unpublished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            "Unpublished"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 (R482Q)-mRFP
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    607..663
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2640..3314
                     /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(2770..2788)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3232..3255
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3448..3569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3485..3504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3539..3558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3576..4031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3713..3734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3925..3944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4058..4162
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4164..4521
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4164..4355
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4187..4207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4372..4507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4556..5350
                     /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             4556..5350
                     /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(4610..4629)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5220..5239
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5538..5557)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5582..5629
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5958..6546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6447..6466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      601 ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
      661 tccgaatggc catggagacc ccgtcccagc ggcgcgccac ccgcagcggg gcgcaggcca
      721 gctccactcc gctgtcgccc acccgcatca cccggctgca ggagaaggag gacctgcagg
      781 agctcaatga tcgcttggcg gtctacatcg accgtgtgcg ctcgctggaa acggagaacg
      841 cagggctgcg ccttcgcatc accgagtctg aagaggtggt cagccgcgag gtgtccggca
      901 tcaaggccgc ctacgaggcc gagctcgggg atgcccgcaa gacccttgac tcagtagcca
      961 aggagcgcgc ccgcctgcag ctggagctga gcaaagtgcg tgaggagttt aaggagctga
     1021 aagcgcgcaa taccaagaag gagggtgacc tgatagctgc tcaggctcgg ctgaaggacc
     1081 tggaggctct gctgaactcc aaggaggccg cactgagcac tgctctcagt gagaagcgca
     1141 cgctggaggg cgagctgcat gatctgcggg gccaggtggc caagcttgag gcagccctag
     1201 gtgaggccaa gaagcaactt caggatgaga tgctgcggcg ggtggatgct gagaacaggc
     1261 tgcagaccat gaaggaggaa ctggacttcc agaagaacat ctacagtgag gagctgcgtg
     1321 agaccaagcg ccgtcatgag acccgactgg tggagattga caatgggaag cagcgtgagt
     1381 ttgagagccg gctggcggat gcgctgcagg aactgcgggc ccagcatgag gaccaggtgg
     1441 agcagtataa gaaggagctg gagaagactt attctgccaa gctggacaat gccaggcagt
     1501 ctgctgagag gaacagcaac ctggtggggg ctgcccacga ggagctgcag cagtcgcgca
     1561 tccgcatcga cagcctctct gcccagctca gccagctcca gaagcagctg gcagccaagg
     1621 aggcgaagct tcgagacctg gaggactcac tggcccgtga gcgggacacc agccggcggc
     1681 tgctggcgga aaaggagcgg gagatggccg agatgcgggc aaggatgcag cagcagctgg
     1741 acgagtacca ggagcttctg gacatcaagc tggccctgga catggagatc cacgcctacc
     1801 gcaagctctt ggagggcgag gaggagaggc tacgcctgtc ccccagccct acctcgcagc
     1861 gcagccgtgg ccgtgcttcc tctcactcat cccagacaca gggtgggggc agcgtcacca
     1921 aaaagcgcaa actggagtcc actgagagcc gcagcagctt ctcacagcac gcacgcacta
     1981 gcgggcgcgt ggccgtggag gaggtggatg aggagggcaa gtttgtccgg ctgcgcaaca
     2041 agtccaatga ggaccagtcc atgggcaatt ggcagatcaa gcgccagaat ggagatgatc
     2101 ccttgctgac ttaccagttc ccaccaaagt tcaccctgaa ggctgggcag gtggtgacga
     2161 tctgggctgc aggagctggg gccacccaca gcccccctac cgacctggtg tggaaggcac
     2221 agaacacctg gggctgcggg aacagcctgc gtacggctct catcaactcc actggggaag
     2281 aagtggccat gcgcaagctg gtgcgctcag tgactgtggt tgaggacgac gaggatgagg
     2341 atggagatga cctgctccat caccaccacg gctcccactg cagcagctcg ggggaccccg
     2401 ctgagtacaa cctgcgctcg cgcaccgtgc tgtgcgggac ctgcgggcag cctgccgaca
     2461 aggcatctgc cagcggctca ggagcccagg tgggcggacc catctcctct ggctcttctg
     2521 cctccagtgt cacggtcact cgcagctacc gcagtgtggg gggcagtggg ggtggcagct
     2581 tcggggacaa tctggtcacc cgctcctacg gtggcggtgg ctctggaggt ggtgggtcca
     2641 tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct
     2701 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2761 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     2821 tgtcccctca gttccagtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2881 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2941 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     3001 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaattcag aagaagacga
     3061 tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga
     3121 tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct
     3181 acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca
     3241 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc
     3301 actccaccgg cgccctgtac aagtaagcgg ccgcgactct agatcataat cagccatacc
     3361 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3421 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3481 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3541 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3601 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3661 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3721 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3781 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3841 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3901 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3961 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4021 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4081 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4141 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4201 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4261 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4321 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4381 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4441 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4501 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4561 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4621 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4681 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4741 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4801 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4861 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4921 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4981 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5041 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5101 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5161 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5221 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5281 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5341 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5401 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5461 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5521 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5581 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5641 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5701 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5761 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5821 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5881 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5941 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     6661 ta
//
