LOCUS       sequence-190907-        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the nuclear envelope (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 6656)
  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: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: mTurquoise2-LaminB
FEATURES             Location/Qualifiers
     source          1..6656
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1369..1410
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    1426..1450
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3248..3272)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3444..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3535..3554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3572..4027)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3709..3730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3921..3940
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4054..4158
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4160..4517
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4160..4351
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4183..4203)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4368..4503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4552..5346
                     /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             4552..5346
                     /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(4606..4625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5216..5235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5534..5553)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5578..5625
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5954..6542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6443..6462
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attgcggtgg taagcctatc
     1381 cctaaccctc tcctcggtct cgattctacg atcgatttcg atcaaacaag tttgtacaaa
     1441 aaagcaggct ccgcggccgc ccccttcacc atggcgactg cgacccccgt gccgccgcgg
     1501 atgggcagcc gcgctggcgg ccccaccacg ccgctgagcc ccacgcgcct gtcgcggctc
     1561 caggagaagg aggagctgcg cgagctcaat gaccggctgg cggtgtacat cgacaaggtg
     1621 cgcagcctgg agacggagaa cagcgcgctg cagctgcagg tgacggagcg cgaggaggtg
     1681 cgcggccgtg agctcaccgg cctcaaggcg ctctacgaga ccgagctggc cgacgcgcga
     1741 cgcgcgctcg acgacacggc ccgcgagcgc gccaagctgc agatcgagct gggcaagtgc
     1801 aaggcggaac acgaccagct gctcctcaac tatgctaaga aggaatctga tcttaatggc
     1861 gcccagatca agcttcgaga atatgaagca gcactgaatt cgaaagatgc agctcttgct
     1921 actgcacttg gtgacaaaaa aagtttagag ggagatttgg aggatctgaa ggatcagatt
     1981 gcccagttgg aagcctcctt agctgcagcc aaaaaacagt tagcagatga aactttactt
     2041 aaagtagatt tggagaatcg ttgtcagagc cttactgagg acttggagtt tcgcaaaagc
     2101 atgtatgaag aggagattaa cgagaccaga aggaagcatg aaacgcgctt ggtagaggtg
     2161 gattctgggc gtcaaattga gtatgagtac aagctggcgc aagcccttca tgagatgaga
     2221 gagcaacatg atgcccaagt gaggctgtat aaggaggagc tggagcagac ttaccatgcc
     2281 aaacttgaga atgccagact gtcatcagag atgaatactt ctactgtcaa cagtgccagg
     2341 gaagaactga tggaaagccg catgagaatt gagagccttt catcccagct ttctaatcta
     2401 cagaaagagt ctagagcatg tttggaaagg attcaagaat tagaggactt gcttgctaaa
     2461 gaaaaagaca actctcgtcg catgctgaca gacaaagaga gagagatggc ggaaataagg
     2521 gatcaaatgc agcaacagct gaatgactat gaacagcttc ttgatgtaaa gttagccctg
     2581 gacatggaaa tcagtgctta caggaaactc ttacaaggcg aagaagagag gttgaagctg
     2641 tctccaagcc cttcttcccg tgtgacagta tcccgagcat cctcaagtcg tagtgtacgt
     2701 acaactagag gaaagcggaa gagggttgat gtggaagaat cagaggcgag tagtagtgtt
     2761 agcatctctc attccgcctc agccactgga aatgtttgca tcgaagaaat tgatgttgat
     2821 gggaaattta tccgcttgaa gaacacttct gaacaggatc aaccaatggg aggctgggag
     2881 atgatcagaa aaattggaga cacatcagtc agttataaat atacctcaag atatgtgctg
     2941 aaggcaggcc agactgttac aatttgggct gcaaacgctg gtgtcacagc cagcccccca
     3001 actgacctca tctggaagaa ccagaactcg tggggcactg gcgaagatgt gaaggttata
     3061 ttgaaaaatt ctcagggaga ggaggttgct caaagaagta cagtctttaa aacaaccata
     3121 cctgaagaag aggaggagga ggaagaagca gctggagtgg ttgttgagga agaacttttc
     3181 caccagcagg gaaccccaag agcatccaat agaagctgtg caattatgta aaagggtggg
     3241 cgcgccgacc cagctttctt gtacaaagtg gttcgatcga accgcggtct agactcgacg
     3301 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     3361 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3421 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3481 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3541 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3601 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3661 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3721 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3781 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3841 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3901 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3961 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4021 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4141 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4441 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4501 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4561 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4621 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4681 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4741 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4801 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4861 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4921 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4981 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5041 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5101 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5161 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5221 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5281 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5341 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5401 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5461 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5521 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5581 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5641 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5701 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5761 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5821 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
