LOCUS       sequence-399875-        5905 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 5905)
  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 5905)
  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: NET33 pEGFP-N2 (597)
FEATURES             Location/Qualifiers
     source          1..5905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             154..948
                     /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             154..948
                     /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(208..227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     818..837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1136..1155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1180..1227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1556..2144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2045..2064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2319..2622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2623..2826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2777..2797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4103..4112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4109..4111,4112..4114,4115..4828)
                     /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(4109..4111,4112..4114,4115..4825)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4154..4175)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4415..4434)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4762..4783
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4951..5072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5042..5061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5079..5534)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5216..5237)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5428..5447
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5561..5665
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5667..5905,1..119)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5667..5858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5690..5710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5875..5905,1..105)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
       61 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
      121 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      181 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      241 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      301 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
      361 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      421 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      481 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      541 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      601 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
      661 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
      721 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
      781 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
      841 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
      901 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
      961 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1021 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     1081 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     1141 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1201 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     1261 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     1321 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     1381 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     1441 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1681 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1981 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2041 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2101 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2161 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2221 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     2281 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2341 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2401 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2461 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2521 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2581 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2641 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2701 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2761 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2821 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagtgat
     2881 ggagaacaaa gctatgtacc tacacaccgt cagcgactgt gacaccagct ccatctgtga
     2941 ggattccttt gatggcagga gcctgtccaa gctgaacctg tgtgaggatg gtccatgtca
     3001 caaacggcgg gcaagcatct gctgtaccca gctggggtcc ctgtcggccc tgaagcatgc
     3061 tgtcctgggg ctctacctgc tggtcttcct gattcttgtg ggcatcttca tcttagcagt
     3121 gtccaggccg cgcagctccc ctgacgacct gaaggccctg actcgcaatg tgaaccggct
     3181 gaatgagagc ttccgggact tgcagctgcg gctgctgcag gctccgctgc aagcggacct
     3241 gacggagcag gtgtggaagg tgcaggacgc gctgcagaac cagtcagact cgttgctggc
     3301 gctggcgggc gcagtgcagc ggctggaggg cgcgctatgg gggctgcagg cgcaggcggt
     3361 gcagaccgag caggcggtgg ccctgctgcg ggaccgcacg ggccagcaga gcgacacggc
     3421 gcagctggag ctctaccagc tgcaggtgga gagcaacagt agccagctgc tgctgaggcg
     3481 ccacgcgggc ctgctggacg ggctggcgcg cagggtgggc atcctgggcg aggagctggc
     3541 cgacgtgggc ggcgtgctgc gcggcctcaa ccacagcctg tcctacgacg tggccctcca
     3601 ccgcacgcgg ctgcaggacc tgcgggtgct ggtgagcaac gccagcgagg acacgcgccg
     3661 cctgcgcctg gcgcacgtag gcatggagct gcagctgaag caggagctgg ccatgctcaa
     3721 cgcggtcacc gaggacctgc gcctcaagga ctgggagcac tccatcgcac tgcggaacat
     3781 ctccctcgcg aaagggccac cgggacccaa aggtgatcag ggggatgaag gaaaggaagg
     3841 caggcctggc atccctggat tgcctggact tcgaggtctg cccggggaga gaggtacccc
     3901 aggattgccc gggcccaagg gcgatgatgg gaagctgggg gccacaggac caatgggcat
     3961 gcgtgggttc aaaggtgacc gaggcccaaa aggagagaaa ggagagaaag gagacagagc
     4021 tggggatgcc agtaaggaca ttctgctggg gccgtgggat atggtgttgg cacaggggcg
     4081 gcccgggatc caccggccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     4141 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     4201 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     4261 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     4321 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4381 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     4441 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     4501 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     4561 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     4621 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4681 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     4741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4801 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     4861 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4921 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5041 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     5101 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5161 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5221 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5281 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5341 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5401 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5461 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5521 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5641 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5701 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5761 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5821 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5881 cccctaactc cgcccagttc cgccc
//
