LOCUS       sequence-101293-        5734 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 5734)
  AUTHORS   Malik P, Korfali N, Srsen V, Lazou V, Batrakou DG, Zuleger N, 
            Kavanagh DM, Wilkie GS, Goldberg MW, Schirmer EC
  TITLE     Cell-specific and lamin-dependent targeting of novel transmembrane 
            proteins in the nuclear envelope.
  JOURNAL   Cell Mol Life Sci. 2010 Apr;67(8):1353-69. doi: 
            10.1007/s00018-010-0257-2. Epub 2010 Jan 21.
  PUBMED    20091084
REFERENCE   2  (bases 1 to 5734)
  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: 
            "10.1007/s00018-010-0257-2"; journalName: "Cell Mol Life Sci"; date:
            "2010-04"; volume: "67"; issue: "8"; pages: "1353-69"
COMMENT     Sequence Label: NET29A.Hs pEGFP-N2 (1181)
FEATURES             Location/Qualifiers
     source          1..5734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1698..1707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1704..1706,1707..1709,1710..2423)
                     /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(1704..1706,1707..1709,1710..2420)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1749..1770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2010..2029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2357..2378
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2546..2667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2583..2602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2637..2656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2674..3129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3023..3042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3156..3260
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3262..3619
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3262..3453
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3285..3305)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3470..3605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3654..4448
                     /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             3654..4448
                     /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(3708..3727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4318..4337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4636..4655)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4680..4727
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5056..5644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5545..5564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      601 gaaccgtcag atccgctagc gctaccggac tcagatctcg acccatgcag cccccgcccc
      661 cgggcccgct gggcgactgc ctgcgggact gggaggatct acagcaggac ttccagaaca
      721 tccaggagac ccatcggctc taccgcctga agctggagga gctgaccaaa cttcagaaca
      781 attgcaccag ctccatcacg cggcagaaga agcggctcca ggagctggcc ctcgccctga
      841 agaaatgcaa accctccctc ccagcagagg ccgagggggc cgcacaggag ctggagaacc
      901 agatgaaaga gcgccaaggc ctcttctttg acatggaggc ctatttgcct aagaagaatg
      961 gattgtacct gagcctggtt ctggggaacg tcaacgtcac gctcctgagc aagcaggcta
     1021 agtttgccta caaggacgag tatgagaagt tcaagctcta cctcaccatc atcctcatcc
     1081 tcatctcctt cacttgccgc ttcctgctca actccagggt gacagatgct gccttcaact
     1141 tcctgctggt ctggtactac tgcaccctga ccatccggga gagcatcctc atcaacaacg
     1201 gctcccggat caaaggctgg tgggtgttcc atcactacgt gtccgccttc ctgtcgggag
     1261 tcatgctgac gtggcccgac ggtctcatgt accagaaatt ccggaaccaa ttcctctcct
     1321 tttccatgta ccagagcttc gtgcagtttc tccagtacta ctaccagagc ggctgcctct
     1381 accgcctgcg ggcgctgggc gagcggcaca ccatggacct cactgtggag ggcttccagt
     1441 cctggatgtg gcggggcctc accttcctgc tgccttttct tttctttgga cacttctggc
     1501 agctttttaa cgcgctgacg ttgttcaacc tggcccagga ccctcagtgc aaggagtggc
     1561 aggtgcttat gtgcggcttt cccttcctcc tccttttcct cggcaatttc ttcaccaccc
     1621 tgagggttgt gcaccacaag tttcacagtc agcggcacgg gagcaagaag gatgcggccg
     1681 ggatccaccg gccggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1741 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1801 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1861 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1921 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1981 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2041 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2101 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2161 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2221 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2281 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2341 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2401 gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac
     2461 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2521 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2641 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2701 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2761 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2821 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2881 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2941 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3001 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3061 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3121 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3181 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3241 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3301 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3421 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3481 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3541 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3601 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3661 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3721 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3781 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3841 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3901 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3961 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4021 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4081 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4141 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4201 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4261 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4321 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4381 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4441 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4501 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4561 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4621 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4681 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4741 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4801 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4861 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4921 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5161 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5701 catgttcttt cctgcgttat cccctgattc tgtg
//
