LOCUS       sequence-342142-        6449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent labeling of the nuclear envelope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  AUTHORS   Taniyama T, Tsuda N, Sueda S
  TITLE     Fluorescent Labeling of the Nuclear Envelope by Localizing Green 
            Fluorescent Protein on the Inner Nuclear Membrane.
  JOURNAL   ACS Chem Biol. 2018 Jun 15;13(6):1463-1469. doi: 
            10.1021/acschembio.8b00219. Epub 2018 May 24.
  PUBMED    29782140
REFERENCE   2  (bases 1 to 6449)
  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"; doi: 
            "10.1021/acschembio.8b00219"; journalName: "ACS Chem Biol"; date: 
            "2018-06-15- 15"; volume: "13"; issue: "6"; pages: "1463-1469"
COMMENT     Sequence Label: pTM-BPL
FEATURES             Location/Qualifiers
     source          1..6449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1096..1105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1210..1233
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1246..1392
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    2153..2274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2164..2183)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2218..2237
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2460..2915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2547..2566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2757..2778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3032..3389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3032..3223
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3055..3075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3240..3375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3440..4234
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3494..3513)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4104..4123
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4298..4346
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4387..4406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4506..4528
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4566..4584)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4652..4756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4757..4825,4826..5617)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4975..4994)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5788..6376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6277..6296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagccac catgggcctc cgggcgctga tgctgtggct gctggcggcg
     1141 gcggggctcg tgcgcgagtc cctgcaagga gagttccaaa ggaagctgta caaggatcga
     1201 ggaagcttgg actacaagga cgatgacgac aaggatccag gaggcgctgt gggccaggac
     1261 acgcaggagg tcatcgtggt gccacactcc ttgcccttta aggtggtggt gatctcagcc
     1321 atcctggccc tggtggtgct caccatcatc tcccttatca tcctcatcat gctttggcag
     1381 aagaagccac gtggaggcgg tgaattcatg ggcatcagca tgctgatctt taagttcccc
     1441 agcgttacct ccacccaaga cctggccgag gccatttatc agattatcaa cgctgatgag
     1501 ttcgtgatag tggcagagga gcagaccagg gccaggggca ggtacaagcg cgagtggtat
     1561 tctcctaagg gagggctctg gtttacatac gtcattaaga attacaatgc agagaagatc
     1621 cctttcctga catttaggtc ctcactggcc gtgaggaagg tgctgtctac atttctgaat
     1681 gttaagatcc gctggcccaa tgacatcgtg tacgggaaaa gaaagattag cggaatcctc
     1741 attgagggca tctcagaggg agcaaacagc actgtcttca tcggtattgg catcgacact
     1801 aatgtcaaat ctctcccaga ggagctgtcc gctacttcac tctatctgga gctgaataga
     1861 gaggttgata atgataaaat gctgaacgag atcataaacg agataaagaa ctacgaaaag
     1921 ataagcgata aggaaataat tgacgaaatc aacgaacacc tctctatcaa ggacaagaag
     1981 gtgaagctca tctccaaatc cgaagaaaga aactgcctgg ctctgttcgt ggactactac
     2041 ggacggctgg tgaccgaatg tggtatcttc gaagtggaag acattctccg ggtcatcgaa
     2101 gaataagcgg ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac
     2161 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2221 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2281 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     2341 aagtaaaacc tctacaaatg tggtaaaatc cgataaggat cgatccgggc tggcgtaata
     2401 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2461 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2521 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2581 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2641 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2701 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2761 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2821 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2881 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     2941 catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa
     3001 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     3061 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3121 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3181 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3301 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3361 gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga
     3421 acttaaggct agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3481 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3541 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3601 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3661 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3721 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3781 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3841 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3901 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3961 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4021 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4081 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4141 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4201 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4261 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     4321 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatccgcg tatggtgcac
     4381 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4441 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4501 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4561 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4621 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4681 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4741 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4801 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4861 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4921 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4981 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5041 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5101 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5161 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5221 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5281 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5341 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5401 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5461 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5521 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5581 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5641 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5701 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5941 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6421 gccttttgct cacatggctc gacagatct
//
