LOCUS       sequence-241955-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lamin A mutant (R435C), C-terminally tagged with mRFP. Mutation 
            associated with Progeria..
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  JOURNAL   Unpublished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            "Unpublished"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 (R435C)-mRFP
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    621..677
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2654..3328
                     /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(2784..2802)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3246..3269
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3462..3583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3499..3518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3553..3572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3590..4045)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3727..3748)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3939..3958
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4072..4176
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4178..4535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4178..4369
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4201..4221)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4386..4521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4570..5364
                     /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             4570..5364
                     /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(4624..4643)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5234..5253
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5552..5571)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5596..5643
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5972..6560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6461..6480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
      661 ccgcgggccc gggatccgaa tggccatgga gaccccgtcc cagcggcgcg ccacccgcag
      721 cggggcgcag gccagctcca ctccgctgtc gcccacccgc atcacccggc tgcaggagaa
      781 ggaggacctg caggagctca atgatcgctt ggcggtctac atcgaccgtg tgcgctcgct
      841 ggaaacggag aacgcagggc tgcgccttcg catcaccgag tctgaagagg tggtcagccg
      901 cgaggtgtcc ggcatcaagg ccgcctacga ggccgagctc ggggatgccc gcaagaccct
      961 tgactcagta gccaaggagc gcgcccgcct gcagctggag ctgagcaaag tgcgtgagga
     1021 gtttaaggag ctgaaagcgc gcaataccaa gaaggagggt gacctgatag ctgctcaggc
     1081 tcggctgaag gacctggagg ctctgctgaa ctccaaggag gccgcactga gcactgctct
     1141 cagtgagaag cgcacgctgg agggcgagct gcatgatctg cggggccagg tggccaagct
     1201 tgaggcagcc ctaggtgagg ccaagaagca acttcaggat gagatgctgc ggcgggtgga
     1261 tgctgagaac aggctgcaga ccatgaagga ggaactggac ttccagaaga acatctacag
     1321 tgaggagctg cgtgagacca agcgccgtca tgagacccga ctggtggaga ttgacaatgg
     1381 gaagcagcgt gagtttgaga gccggctggc ggatgcgctg caggaactgc gggcccagca
     1441 tgaggaccag gtggagcagt ataagaagga gctggagaag acttattctg ccaagctgga
     1501 caatgccagg cagtctgctg agaggaacag caacctggtg ggggctgccc acgaggagct
     1561 gcagcagtcg cgcatccgca tcgacagcct ctctgcccag ctcagccagc tccagaagca
     1621 gctggcagcc aaggaggcga agcttcgaga cctggaggac tcactggccc gtgagcggga
     1681 caccagccgg cggctgctgg cggaaaagga gcgggagatg gccgagatgc gggcaaggat
     1741 gcagcagcag ctggacgagt accaggagct tctggacatc aagctggccc tggacatgga
     1801 gatccacgcc taccgcaagc tcttggaggg cgaggaggag aggctacgcc tgtcccccag
     1861 ccctacctcg cagcgcagcc gtggccgtgc ttcctctcac tcatcccaga cacagggtgg
     1921 gggcagcgtc accaaaaagc gcaaactgga gtccactgag agccgcagca gcttctcaca
     1981 gcacgcatgc actagcgggc gcgtggccgt ggaggaggtg gatgaggagg gcaagtttgt
     2041 ccggctgcgc aacaagtcca atgaggacca gtccatgggc aattggcaga tcaagcgcca
     2101 gaatggagat gatcccttgc tgacttaccg gttcccacca aagttcaccc tgaaggctgg
     2161 gcaggtggtg acgatctggg ctgcaggagc tggggccacc cacagccccc ctaccgacct
     2221 ggtgtggaag gcacagaaca cctggggctg cgggaacagc ctgcgtacgg ctctcatcaa
     2281 ctccactggg gaagaagtgg ccatgcgcaa gctggtgcgc tcagtgactg tggttgagga
     2341 cgacgaggat gaggatggag atgacctgct ccatcaccac cacggctccc actgcagcag
     2401 ctcgggggac cccgctgagt acaacctgcg ctcgcgcacc gtgctgtgcg ggacctgcgg
     2461 gcagcctgcc gacaaggcat ctgccagcgg ctcaggagcc caggtgggcg gacccatctc
     2521 ctctggctct tctgcctcca gtgtcacggt cactcgcagc taccgcagtg tggggggcag
     2581 tgggggtggc agcttcgggg acaatctggt cacccgctcc tacggtggcg gtggctctgg
     2641 aggtggtggg tccatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt
     2701 gcgcatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     2761 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
     2821 cgcctgggac atcctgtccc ctcagttcca gtacggctcc aaggcctacg tgaagcaccc
     2881 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     2941 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3001 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3061 tcagaagaag acgatgggct gggaggcctc caccgagcgg atgtaccccg aggacggcgc
     3121 cctgaagggc gagatcaaga tgaggctgaa gctgaaggac ggcggccact acgacgccga
     3181 ggtcaagacc acctacatgg ccaagaagcc cgtgcagctg cccggcgcct acaagaccga
     3241 catcaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     3301 cgccgagggc cgccactcca ccggcgccct gtacaagtaa gcggccgcga ctctagatca
     3361 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3421 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3481 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3541 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3601 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3661 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3721 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3781 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3841 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3901 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3961 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4021 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4141 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4201 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4261 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4321 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4381 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4441 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4501 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4561 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4621 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4681 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4741 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4801 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4861 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4921 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4981 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5041 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5101 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5161 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5221 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5281 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5341 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5401 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5461 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5521 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5581 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5641 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5701 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5761 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5941 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6541 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6601 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6661 tt
//
