LOCUS       sequence-274824-        6054 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 6054)
  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 6054)
  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: LAP2 Full I pAcGFP-N1 monomeric GFP (1317)
FEATURES             Location/Qualifiers
     source          1..6054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1967..2686
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2808..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2845..2864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2899..2918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2936..3391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3073..3094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3285..3304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3418..3522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3524..3881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3524..3715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3547..3567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3732..3867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3916..4710
                     /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             3916..4710
                     /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(3970..3989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4580..4599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4898..4917)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4942..4989
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5318..5906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5807..5826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      541 gtgaaccgtc agatccgcta gcgctaccgg actcagatcc atgccggagt tcctagagga
      601 cccttcggtc ctgaccaaag acaagttgaa gagcgagttg gtcgccaaca acgtgacgct
      661 cccggccggc gagcagcgca aggacgtgta cgtgcagctc tacctgcagc acctcacggc
      721 gcgcaaccgg ccgccgctcg ccgcgggagc caacagcaaa gggccgcccg acttctcgag
      781 cgacgaggag cgcgagccca ccccagtgct tggctccggg gcctccgtgg gtcgcggccg
      841 cggcgccgtc ggcaggaaag ccacaaagaa aactgataag cccaggccag aagataaaga
      901 tgatctggat gtgacagagc tctctaacga agaacttctg gaacagcttg tgagatatgg
      961 agtgaatcct ggtcccattg tgggaaccac caggaagctg tatgagaaga agctgttgaa
     1021 gctgagggaa cagggagcag aatcgagatc ctctactcct ctcccaacag tctcttcctc
     1081 tgcagaaaac acgaggcaga atggaagtaa tgactctgac agatacagtg acaatgacga
     1141 agactctaaa atagagctca agcttgagaa gagagagccg ctaaagggca gagcaaagac
     1201 tccagtaaca ctcaagcaaa ggaggattga gcacaatcag agctattctg aagctggagt
     1261 aactgagact gaatggacaa gtggatcttc aaaaggcgga cctctgcagg cattaactag
     1321 ggagtccacg agagggtcga gaagaactcc aaggagaagg gtggaaccct cacagcattt
     1381 tcgtgtagat ggtgcagtaa tttcagagag tactcccata gctgagacta taaaggcttc
     1441 cagcaacgac tccttagtgg ccaataggtt gactggaaat ttcaagcatg catcttctat
     1501 tctgccaatc actgaattct cagacataac cagaagaaca ccaaagaaac cattgacaag
     1561 agctgaagtg ggagaaaaaa cagaggaaag aagagtagaa agggatattc tgaaggaaat
     1621 gttcccgtat gaagcctcta ctccaaccgg aattagtgct agttgccgca gaccaatcaa
     1681 aggtgctgcc ggccggccgc tcgagctcag tgacttcagg atggaagaat cgttctcatc
     1741 taagtacgtc ccgaagtacg ttcccttggc agacgtcaag tcggaaaaga caaagaaggg
     1801 acgctccgtt cccatgtgga taaaaatgct gctgtttgct ctcgtggcgg gttttttgtt
     1861 tttggtctat caagctatgg aaaccaacca aggaaacccc ttcactaatt ttcttcaaga
     1921 tactaaaata tccaactggg taccgcgggc ccgggatcca ccggtcatgg tgagcaaggg
     1981 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     2041 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2101 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     2161 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     2221 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     2281 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     2341 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     2401 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     2461 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     2521 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     2581 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     2641 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtgagcgg ccgcgactct
     2701 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2761 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2941 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3001 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3061 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3121 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3181 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3241 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3301 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3361 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3421 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3541 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3781 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3901 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3961 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4021 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4081 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4141 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4201 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4261 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4321 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4381 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4441 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4501 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4561 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4621 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4681 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4741 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4801 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4861 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4921 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4981 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5041 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5101 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5161 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5221 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5281 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5341 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5401 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5461 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5521 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5581 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5641 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5701 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5761 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5821 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5881 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5941 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6001 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc atta
//
