LOCUS       sequence-94052-e        7108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LBR-EGFP in mammalian cells (CMV promoter). Also suitable 
            for in vitro transcription (T7 promoter) for injecting mRNA into 
            oocytes. This LBR contains N-terminal domain only..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  AUTHORS   Inoue A, Zhang Y
  TITLE     Nucleosome assembly is required for nuclear pore complex assembly in
            mouse zygotes.
  JOURNAL   Nat Struct Mol Biol. 2014 Jul;21(7):609-16. doi: 10.1038/nsmb.2839. 
            Epub 2014 Jun 8.
  PUBMED    24908396
REFERENCE   2  (bases 1 to 7108)
  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: "10.1038/nsmb.2839";
            journalName: "Nat Struct Mol Biol"; date: "2014-07"; volume: "21"; 
            issue: "7"; pages: "609-16"
COMMENT     Sequence Label: pcDNA-LBR-EGFP-polyA
FEATURES             Location/Qualifiers
     source          1..7108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1674..1683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1680..1682,1683..1685,1686..2399)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             1680..2396
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1725..1746)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1986..2005)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2333..2354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2611..2640
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2656..2673
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2696..2713)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2702..2926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2972..3400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3059..3078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3269..3290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3409..3429)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3414..3744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3595..3730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3657..3676
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3811..4605
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3865..3884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4475..4494
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4781..4902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4818..4837)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4872..4891
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4951..4967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4951..4967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4964..4986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4975..4991
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4999..5005,5006..5023,5024..5029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5044..5065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5182..5199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5433..5452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6112..6903,6904..6972))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6735..6754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6973..7077)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttcga attcatgcca agtaggaagt ttgttgaggg tgaagtggtc agaggccgat
      961 ggcctgggag ctccctttat tatgaagtag agattctgag ccacgacaac aaatcccagc
     1021 tctacactgt aaaatacaaa gatggcaccg aactggaact gaaggagagt gatatcaagc
     1081 ctttaaaatc ctttaagcaa agaaaaagtg gctcgatttc cagctctccc tctcgacgcc
     1141 gtggcagccg cagccgcagc cgctcaagat cacgttcccg atccccgggc agagcaccga
     1201 aaggctccag aagatccgtc tctgcttccc atgagggtga cgtgaaggag aagaaggaga
     1261 aggagatgag gagggaaatt ctgcaagtta agctgactcc gcttgtgttg aagccgttcg
     1321 gaaacagcgt cagcgtgtac aacggggagc ccgaacacat ggagaagaac gctacgccgt
     1381 ataaagacaa gcaggaaaga atcattttgt caacagaaga cagatatata gttacacagt
     1441 acagccttcg tcccaggaga gaagaggtca aagccaaaga aatagagtct gaggaacaaa
     1501 accttgtcac caaaggaccc gcacctttgg gaacctttca agtgaccact ccacagagga
     1561 aggacttgga gtttggagga gtacctggtg cggtcctcat catgctgggc ctgcccgcct
     1621 gcgtcctctt gctgctgctg cagtgcaggc agaaggaccc ggatccaccg gtcgccacca
     1681 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1741 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1801 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1861 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1921 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1981 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2041 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2101 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2161 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2221 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2281 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2341 tgctggagtt cgtgaccgcc gccgggatca ctcacggcat ggacgagctg tacaagtaag
     2401 cggccgctcg actatgggca ccaaagaacc tgtaaacgtt atctttttaa attgaatgtg
     2461 cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2521 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaatacgc atcatacgct
     2581 gtggcgtacc tcgagtctag agggcccttc gaacaaaaac tcatctcaga agaggatctg
     2641 aatatgcata ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg
     2701 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2761 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2821 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2881 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2941 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3001 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3061 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3121 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3181 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3241 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3301 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     3361 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3421 agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat
     3481 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3541 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3601 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3661 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3721 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3781 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3841 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3961 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4321 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4621 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4681 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4741 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4801 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4861 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4921 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4981 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5041 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5101 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7081 atttccccga aaagtgccac ctgacgtc
//
