LOCUS       sequence-241962-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lamin A mutant (G465D), 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  JOURNAL   Unpublished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            "Unpublished"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 (G465D)-mRFP
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..601
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    642..698
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2675..3349
                     /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(2805..2823)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3267..3290
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3483..3604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3520..3539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3574..3593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3611..4066)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3748..3769)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3960..3979
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4093..4197
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4199..4556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4199..4390
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4222..4242)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4407..4542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4591..5385
                     /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             4591..5385
                     /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(4645..4664)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5255..5274
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5573..5592)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5617..5664
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5993..6581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6482..6501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      601 tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga gctcaagctt
      661 cgaattctgc agtcgacggt accgcgggcc cgggatccga atggccatgg agaccccgtc
      721 ccagcggcgc gccacccgca gcggggcgca ggccagctcc actccgctgt cgcccacccg
      781 catcacccgg ctgcaggaga aggaggacct gcaggagctc aatgatcgct tggcggtcta
      841 catcgaccgt gtgcgctcgc tggaaacgga gaacgcaggg ctgcgccttc gcatcaccga
      901 gtctgaagag gtggtcagcc gcgaggtgtc cggcatcaag gccgcctacg aggccgagct
      961 cggggatgcc cgcaagaccc ttgactcagt agccaaggag cgcgcccgcc tgcagctgga
     1021 gctgagcaaa gtgcgtgagg agtttaagga gctgaaagcg cgcaatacca agaaggaggg
     1081 tgacctgata gctgctcagg ctcggctgaa ggacctggag gctctgctga actccaagga
     1141 ggccgcactg agcactgctc tcagtgagaa gcgcacgctg gagggcgagc tgcatgatct
     1201 gcggggccag gtggccaagc ttgaggcagc cctaggtgag gccaagaagc aacttcagga
     1261 tgagatgctg cggcgggtgg atgctgagaa caggctgcag accatgaagg aggaactgga
     1321 cttccagaag aacatctaca gtgaggagct gcgtgagacc aagcgccgtc atgagacccg
     1381 actggtggag attgacaatg ggaagcagcg tgagtttgag agccggctgg cggatgcgct
     1441 gcaggaactg cgggcccagc atgaggacca ggtggagcag tataagaagg agctggagaa
     1501 gacttattct gccaagctgg acaatgccag gcagtctgct gagaggaaca gcaacctggt
     1561 gggggctgcc cacgaggagc tgcagcagtc gcgcatccgc atcgacagcc tctctgccca
     1621 gctcagccag ctccagaagc agctggcagc caaggaggcg aagcttcgag acctggagga
     1681 ctcactggcc cgtgagcggg acaccagccg gcggctgctg gcggaaaagg agcgggagat
     1741 ggccgagatg cgggcaagga tgcagcagca gctggacgag taccaggagc ttctggacat
     1801 caagctggcc ctggacatgg agatccacgc ctaccgcaag ctcttggagg gcgaggagga
     1861 gaggctacgc ctgtccccca gccctacctc gcagcgcagc cgtggccgtg cttcctctca
     1921 ctcatcccag acacagggtg ggggcagcgt caccaaaaag cgcaaactgg agtccactga
     1981 gagccgcagc agcttctcac agcacgcacg cactagcggg cgcgtggccg tggaggaggt
     2041 ggatgaggag ggcaagtttg tccggctgcg caacaagtcc aatgaggacc agtccatgga
     2101 caattggcag atcaagcgcc agaatggaga tgatcccttg ctgacttacc ggttcccacc
     2161 aaagttcacc ctgaaggctg ggcaggtggt gacgatctgg gctgcaggag ctggggccac
     2221 ccacagcccc cctaccgacc tggtgtggaa ggcacagaac acctggggct gcgggaacag
     2281 cctgcgtacg gctctcatca actccactgg ggaagaagtg gccatgcgca agctggtgcg
     2341 ctcagtgact gtggttgagg acgacgagga tgaggatgga gatgacctgc tccatcacca
     2401 ccacggctcc cactgcagca gctcggggga ccccgctgag tacaacctgc gctcgcgcac
     2461 cgtgctgtgc gggacctgcg ggcagcctgc cgacaaggca tctgccagcg gctcaggagc
     2521 ccaggtgggc ggacccatct cctctggctc ttctgcctcc agtgtcacgg tcactcgcag
     2581 ctaccgcagt gtggggggca gtgggggtgg cagcttcggg gacaatctgg tcacccgctc
     2641 ctacggtggc ggtggctctg gaggtggtgg gtccatggcc tcctccgagg acgtcatcaa
     2701 ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat
     2761 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     2821 caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttcc agtacggctc
     2881 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     2941 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     3001 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     3061 cccctccgac ggccccgtaa ttcagaagaa gacgatgggc tgggaggcct ccaccgagcg
     3121 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag atgaggctga agctgaagga
     3181 cggcggccac tacgacgccg aggtcaagac cacctacatg gccaagaagc ccgtgcagct
     3241 gcccggcgcc tacaagaccg acatcaagct ggacatcacc tcccacaacg aggactacac
     3301 catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcgccc tgtacaagta
     3361 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3421 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3481 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3541 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3601 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3661 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3721 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3781 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3841 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3901 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3961 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4021 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4081 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4141 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4201 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4261 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4321 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4381 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4441 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4501 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4561 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4621 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4681 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4741 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4801 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4861 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4921 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4981 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5041 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5101 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5161 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5221 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5281 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5341 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5401 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5461 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5521 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5581 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5641 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5701 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5761 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5821 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6661 ct
//
