LOCUS       sequence-339427-        6562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear envelope protein LBR with a C-terminal mCherry
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6562)
  AUTHORS   Ferrandiz N, Downie L, Starling GP, Royle SJ
  TITLE     Endomembranes promote chromosome missegregation by ensheathing 
            misaligned chromosomes.
  JOURNAL   J Cell Biol. 2022 Jun 6;221(6). pii: 213189. doi: 
            10.1083/jcb.202203021. Epub 2022 Apr 29.
  PUBMED    35486148
REFERENCE   2  (bases 1 to 6562)
  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: "2022-06-6"; volume: "221(6). pii: 213189. doi"; 
            pages: " 10.1083/jcb.202203021. Epub 2022 Apr 29"
COMMENT     Sequence Label: LBR-mCherry
FEATURES             Location/Qualifiers
     source          1..6562
                     /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"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2512..2521
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2518..3228
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2663..2681)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2931..2950
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3350..3471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3387..3406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3441..3460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3478..3933)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3615..3636)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3827..3846
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3960..4064
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4066..4423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4066..4257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4089..4109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4274..4409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4458..5252
                     /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             4458..5252
                     /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(4512..4531)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5122..5141
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5440..5459)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5484..5531
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5860..6448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6349..6368
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc catgccaagt
      661 aggaaatttg ccgatggtga agtggtaaga ggtcgatggc ctgggagttc actttattat
      721 gaagtagaaa ttctgagcca cgacagcacc tcccagcttt acactgtgaa gtataaagat
      781 ggaacagagc ttgaattgaa agagaatgat attaagcctt taacttcctt taggcaaagg
      841 aaaggtggct caacttccag ttccccttcc agacgccgag ggagtcgatc aaggtcacgc
      901 tcccgatccc ctggtcgacc acctaaaagt gcccgccgat ctgcttctgc ttcccaccag
      961 gccgacatta aggaagcaag gagggaagtg gaagttaaat tgactccgct gattctgaag
     1021 ccatttggaa atagcatcag cagatataat ggggagcctg agcatattga gagaaatgac
     1081 gcacctcata aaaatacaca ggaaaaattc agtttgtcac aagaaagcag ttacatagca
     1141 acacagtata gccttcgtcc aagaagagaa gaagtcaaat taaaagaaat agattctaag
     1201 gaagaaaaat acgttgcaaa agaactggca gtgagaacct ttgaagtgac ccccatccgg
     1261 gcaaaggact tggagtttgg aggagtacct ggtgtgtttc tcatcatgtt tggcctgcct
     1321 gtgttcctct tcctgttgct gttgatgtgt aaacagaaag atcccagtct tctgaatttc
     1381 cctcctcctt tgccagcttt gtatgagtta tgggaaacca gagtatttgg ggtctacctc
     1441 ctgtggtttt tgattcaagt cctgttctac ctactgccaa ttggaaaggt tgtagaagga
     1501 acgcctctta ttgatggaag aagactcaag tatagattaa atggattcta tgcttttatc
     1561 ctgacatctg cagtcatcgg aacatctctc ttccagggcg tagagtttca ttacgtgtac
     1621 agtcattttc ttcagtttgc acttgcggcc actgtttttt gtgtggtctt gagtgtgtat
     1681 ctctacatgc gctctttgaa agcgccccgg aatgacctgt cgcctgccag ctctggaaat
     1741 gctgtctatg atttcttcat tggccgtgaa ttaaaccctc gaattggtac ttttgatctc
     1801 aaatactttt gtgaattgcg ccccggattg attggatggg tggttattaa cttggtgatg
     1861 cttttggctg aaatgaaaat acaggaccgc gctgttccat ccttggccat gattttagtt
     1921 aatagtttcc agcttctcta tgtggtggat gctctctgga atgaggaagc gttgttgacg
     1981 accatggaca tcatccacga tggatttgga ttcatgctgg cttttggaga cttggtgtgg
     2041 gttcccttta tttacagctt ccaagccttt tatttagtca gtcatccaaa tgaagtgtct
     2101 tggccaatgg cttctctaat tattgttctg aaactttgtg gttatgtaat cttccgaggt
     2161 gcaaattctc agaaaaatgc attccggaaa aatcccagtg atccaaagct tgcacattta
     2221 aaaaccattc atacttcaac gggaaaaaat cttctagttt ctggatggtg gggctttgtt
     2281 cgccacccca attacttggg tgatctcatc atggccttgg cgtggtccct cccatgtggt
     2341 tttaaccaca ttctgcctta tttctacata atttatttca ccatgttgct tgtccaccga
     2401 gaagctcgtg acgagtacca ctgtaagaag aaatacggcg tggcttggga aaagtactgt
     2461 cagcgtgtgc cctaccgtat atttccatac atctacacgg atccaccggt cgccaccatg
     2521 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2581 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2641 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2701 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2761 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2821 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2881 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2941 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     3001 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     3061 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     3121 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     3181 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc ggccgcgact
     3241 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3301 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3361 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3421 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3481 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3541 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3601 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3661 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3721 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3781 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3841 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3901 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3961 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4021 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4081 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4141 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4201 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4261 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4321 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4381 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4441 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4501 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4561 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4621 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4681 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4741 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4801 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4861 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4921 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4981 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5041 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5101 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5161 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5221 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5281 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5341 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5401 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5461 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5521 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5581 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5641 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5701 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5881 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5941 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6001 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6061 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6121 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6181 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6241 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6301 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6361 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6421 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6481 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6541 taaccgtatt accgccatgc at
//
