LOCUS       sequence-462158-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HaloTag-tagged LaminA to visualize the nuclear lamina.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  AUTHORS   Korsten G, Osinga M, Pelle RA, Serweta AK, Hoogenberg B, Kampinga 
            HH, Kapitein LC
  TITLE     Nuclear poly-glutamine aggregates rupture the nuclear envelope and 
            hinder its repair.
  JOURNAL   J Cell Biol. 2024 Nov 4;223(11):e202307142. doi: 
            10.1083/jcb.202307142. Epub 2024 Aug 16.
  PUBMED    39150509
REFERENCE   2  (bases 1 to 6890)
  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: "J Cell 
            Biol."; date: "2024-11-4"; volume: "223(11):e202307142. doi"; pages:
            " 10.1083/jcb.202307142. Epub 2024 Aug 16"
COMMENT     Sequence Label: HaloTag-LaminA
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             597..1484
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     polyA_signal    3678..3799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3715..3734)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3769..3788
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3806..4261)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3943..3964)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4155..4174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4288..4392
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4394..4751
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4394..4585
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4417..4437)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4602..4737
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4786..5580
                     /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             4786..5580
                     /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(4840..4859)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5450..5469
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5768..5787)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5812..5859
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6188..6776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6677..6696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc
      661 actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga
      721 cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg
      781 ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg
      841 accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg
      901 tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg
      961 tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag
     1021 aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca
     1081 tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg
     1141 aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt
     1201 ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg
     1261 aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc
     1321 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
     1381 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
     1441 gcagcgagat cgcgcgctgg ctgtccacgc tcgagatttc cggcagatct cgagctcaag
     1501 cttcgaattc tgcagtcgac ggtaccgcgg gcagcgctgc caacctgccg gccatggaga
     1561 ccccgtccca gcggcgcgcc acccgcagcg gggcgcaggc cagctccact ccgctgtcgc
     1621 ccacccgcat cacccggctg caggagaagg aggacctgca ggagctcaat gatcgcttgg
     1681 cggtctacat cgaccgtgtg cgctcgctgg aaacggagaa cgcagggctg cgccttcgca
     1741 tcaccgagtc tgaagaggtg gtcagccgcg aggtgtccgg catcaaggcc gcctacgagg
     1801 ccgagctcgg ggatgcccgc aagacccttg actcagtagc caaggagcgc gcccgcctgc
     1861 agctggagct gagcaaagtg cgtgaggagt ttaaggagct gaaagcgcgc aataccaaga
     1921 aggagggtga cctgatagct gctcaggctc ggctgaagga cctggaggct ctgctgaact
     1981 ccaaggaggc cgcactgagc actgctctca gtgagaagcg cacgctggag ggcgagctgc
     2041 atgatctgcg gggccaggtg gccaagcttg aggcagccct aggtgaggcc aagaagcaac
     2101 ttcaggatga gatgctgcgg cgggtggatg ctgagaacag gctgcagacc atgaaggagg
     2161 aactggactt ccagaagaac atctacagtg aggagctgcg tgagaccaag cgccgtcatg
     2221 agacccgact ggtggagatt gacaatggga agcagcgtga gtttgagagc cggctggcgg
     2281 atgcgctgca ggaactgcgg gcccagcatg aggaccaggt ggagcagtat aagaaggagc
     2341 tggagaagac ttattctgcc aagctggaca atgccaggca gtctgctgag aggaacagca
     2401 acctggtggg ggctgcccac gaggagctgc agcagtcgcg catccgcatc gacagcctct
     2461 ctgcccagct cagccagctc cagaagcagc tggcagccaa ggaggcgaag cttcgagacc
     2521 tggaggactc actggcccgt gagcgggaca ccagccggcg gctgctggcg gaaaaggagc
     2581 gggagatggc cgagatgcgg gcaaggatgc agcagcagct ggacgagtac caggagcttc
     2641 tggacatcaa gctggccctg gacatggaga tccacgccta ccgcaagctc ttggagggcg
     2701 aggaggagag gctacgcctg tcccccagcc ctacctcgca gcgcagccgt ggccgtgctt
     2761 cctctcactc atcccagaca cagggtgggg gcagcgtcac caaaaagcgc aaactggagt
     2821 ccactgagag ccgcagcagc ttctcacagc acgcacgcac tagcgggcgc gtggccgtgg
     2881 aggaggtgga tgaggagggc aagtttgtcc ggctgcgcaa caagtccaat gaggaccagt
     2941 ccatgggcaa ttggcagatc aagcgccaga atggagatga tcccttgctg acttaccggt
     3001 tcccaccaaa gttcaccctg aaggctgggc aggtggtgac gatctgggct gcaggagctg
     3061 gggccaccca cagcccccct accgacctgg tgtggaaggc acagaacacc tggggctgcg
     3121 ggaacagcct gcgtacggct ctcatcaact ccactgggga agaagtggcc atgcgcaagc
     3181 tggtgcgctc agtgactgtg gttgaggacg acgaggatga ggatggagat gacctgctcc
     3241 atcaccacca cggctcccac tgcagcagct cgggggaccc cgctgagtac aacctgcgct
     3301 cgcgcaccgt gctgtgcggg acctgcgggc agcctgccga caaggcatct gccagcggct
     3361 caggagccca ggtgggcgga cccatctcct ctggctcttc tgcctccagt gtcacggtca
     3421 ctcgcagcta ccgcagtgtg gggggcagtg ggggtggcag cttcggggac aatctggtca
     3481 cccgctccta cctcctgggc aactccagcc cccgaaccca gagcccccag aactgcagca
     3541 tcatgtaagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3601 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3661 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3721 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3781 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3841 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3901 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3961 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4021 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4081 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4141 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4201 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4261 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4321 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4381 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4441 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4501 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4561 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4621 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4681 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4741 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4801 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4861 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4921 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4981 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5041 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5101 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5161 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5221 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5281 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5341 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5401 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5461 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5521 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5581 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5641 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5701 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5761 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5821 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5881 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5941 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6001 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6061 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6121 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6181 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6241 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6301 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6361 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6421 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6481 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6541 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6601 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6661 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6721 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6781 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6841 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
