LOCUS       sequence-375340-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of nuclear envelope transmembrane protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  AUTHORS   Zuleger N, Kelly DA, Richardson AC, Kerr AR, Goldberg MW, Goryachev 
            AB, Schirmer EC
  TITLE     System analysis shows distinct mechanisms and common principles of 
            nuclear envelope protein dynamics.
  JOURNAL   J Cell Biol. 2011 Apr 4;193(1):109-23. doi: 10.1083/jcb.201009068. 
            Epub 2011 Mar 28.
  PUBMED    21444689
REFERENCE   2  (bases 1 to 6524)
  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: 
            "10.1083/jcb.201009068"; journalName: "J Cell Biol"; date: 
            "2011-04-4- 4"; volume: "193"; issue: "1"; pages: "109-23"
COMMENT     Sequence Label: LBR pEGFP-N2 (646)
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..324
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     275..295
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2220..2229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2226..2228,2229..2231,2232..2945)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2226..2228,2229..2231,2232..2942)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2271..2292)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2532..2551)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2879..2900
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3068..3189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3105..3124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3159..3178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3196..3651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3333..3354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3545..3564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3678..3782
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3784..4141
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3784..3975
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3807..3827)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3992..4127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4176..4970
                     /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             4176..4970
                     /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(4230..4249)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4840..4859
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5158..5177)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5202..5249
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5578..6166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6067..6086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(6341..6524,1..120)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
       61 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      121 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      181 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      241 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      301 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gctatgccaa
      361 gtaggaaatt tgccgatggt gaagtggtaa gaggtcgatg gcctgggagt tcactttatt
      421 atgaagtaga aattctgagc cacgacagca cctcccagct ttacactgtg aagtataaag
      481 atggaacaga gcttgaattg aaagagaatg atattaagcc tttaacttcc tttaggcaaa
      541 ggaaaggtgg ctcaacttcc agttcccctt ccagacgccg agggagtcga tcaaggtcac
      601 gctcccgatc ccctggtcga ccacctaaaa gtgcccgccg atctgcttct gcttcccacc
      661 aggccgacat taaggaagca aggagggaag tggaagttaa attgactccg ctgattctga
      721 agccatttgg aaatagcatc agcagatata atggggagcc tgagcatatt gagagaaatg
      781 acgcacctca taaaaataca caggaaaaat tcagtttgtc acaagaaagc agttacatag
      841 caacacagta tagccttcgt ccaagaagag aagaagtcaa attaaaagaa atagattcta
      901 aggaagaaaa atacgttgca aaagaactgg cagtgagaac ctttgaagtg acccccatcc
      961 gggcaaagga cttggagttt ggaggagtac ctggtgtgtt tctcatcatg tttggcctgc
     1021 ctgtgttcct cttcctgttg ctgttgatgt gtaaacagaa agatcccagt cttctgaatt
     1081 tccctcctcc tttgccagct ttgtatgagt tatgggaaac cagagtattt ggggtctacc
     1141 tcctgtggtt tttgattcaa gtcctgttct acctactgcc aattggaaag gttgtagaag
     1201 gaacgcctct tattgatgga agaagactca agtatagatt aaatggattc tatgctttta
     1261 tcctgacatc tgcagtcatc ggaacatctc tcttccaggg cgtagagttt cattacgtgt
     1321 acagtcattt tcttcagttt gcacttgcgg ccactgtttt ttgtgtggtc ttgagtgtgt
     1381 atctctacat gcgctctttg aaagcgcccc ggaatgacct gtcgcctgcc agctctggaa
     1441 atgctgtcta tgatttcttc attggccgtg aattaaaccc tcgaattggt acttttgatc
     1501 tcaaatactt ttgtgaattg cgccccggat tgattggatg ggtggttatt aacttggtga
     1561 tgcttttggc tgaaatgaaa atacaggacc gcgctgttcc atccttggcc atgattttag
     1621 ttaatagttt ccagcttctc tatgtggtgg atgctctctg gaatgaggaa gcgttgttga
     1681 cgaccatgga catcatccac gatggatttg gattcatgct ggcttttgga gacttggtgt
     1741 gggttccctt tatttacagc ttccaagcct tttatttagt cagtcatcca aatgaagtgt
     1801 cttggccaat ggcttctcta attattgttc tgaaactttg tggttatgta atcttccgag
     1861 gtgcaaattc tcagaaaaat gcattccgga aaaatcccag tgatccaaag cttgcacatt
     1921 taaaaaccat tcatacttca acgggaaaaa atcttctagt ttctggatgg tggggctttg
     1981 ttcgccaccc caattacttg ggtgatctca tcatggcctt ggcgtggtcc ctcccatgtg
     2041 gttttaacca cattctgcct tatttctaca taatttattt caccatgttg cttgtccacc
     2101 gagaagctcg tgacgagtac cactgtaaga agaaatacgg cgtggcttgg gaaaagtact
     2161 gtcagcgtgt gccctaccgt atatttccat acatctacgg aaggatccac cggccggtcg
     2221 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2281 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2341 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2401 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2461 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2521 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2581 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2641 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2701 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2761 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2821 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2881 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2941 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3001 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3061 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3121 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3181 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3241 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3301 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3361 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3421 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3481 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3541 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3601 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3781 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3901 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3961 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4021 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4081 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4141 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4201 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4261 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4321 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4381 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4441 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4501 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4561 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4621 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4681 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4741 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4801 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4861 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4921 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4981 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5041 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5101 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5161 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5221 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5281 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5341 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5401 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5461 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5521 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5581 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5641 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5701 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5761 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5821 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5881 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5941 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6001 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6061 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6121 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6181 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6241 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     6301 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6361 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6421 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6481 aaactgccca cttggcagta catcaagtgt atcatatgcc aagt
//
