LOCUS       sequence-94053-e        7062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LBR-mRFP1 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 7062)
  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 7062)
  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-mRFP1-polyA
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        243..622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        623..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     777..797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     871..890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        871..889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1676..2353
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1676..2353
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1806..1824)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2074..2093
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2268..2291
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2565..2594
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2610..2627
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2650..2667)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2656..2880
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2926..3354
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3013..3032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3223..3244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3363..3383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3368..3698
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3549..3684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3611..3630
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3765..4559
                     /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(3819..3838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4429..4448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4735..4856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4772..4791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4826..4845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4905..4921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4905..4921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4918..4940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4929..4945
                     /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(4953..4959,4960..4977,4978..4983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4998..5019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5136..5153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5307..5895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5387..5406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6066..6857,6858..6926))
                     /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     6689..6708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6927..7031)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tncnacgtga cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      181 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      241 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      301 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      361 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      421 gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact tggcagtaca
      481 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      541 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      601 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      661 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      721 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      781 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
      841 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
      901 tggctagtta agcttcgaat tcatgccaag taggaagttt gttgagggtg aagtggtcag
      961 aggccgatgg cctgggagct ccctttatta tgaagtagag attctgagcc acgacaacaa
     1021 atcccagctc tacactgtaa aatacaaaga tggcaccgaa ctggaactga aggagagtga
     1081 tatcaagcct ttaaaatcct ttaagcaaag aaaaagtggc tcgatttcca gctctccctc
     1141 tcgacgccgt ggcagccgca gccgcagccg ctcaagatca cgttcccgat ccccgggcag
     1201 agcaccgaaa ggctccagaa gatccgtctc tgcttcccat gagggtgacg tgaaggagaa
     1261 gaaggagaag gagatgagga gggaaattct gcaagttaag ctgactccgc ttgtgttgaa
     1321 gccgttcgga aacagcgtca gcgtgtacaa cggggagccc gaacacatgg agaagaacgc
     1381 tacgccgtat aaagacaagc aggaaagaat cattttgtca acagaagaca gatatatagt
     1441 tacacagtac agccttcgtc ccaggagaga agaggtcaaa gccaaagaaa tagagtctga
     1501 ggaacaaaac cttgtcacca aaggacccgc acctttggga acctttcaag tgaccactcc
     1561 acagaggaag gacttggagt ttggaggagt acctggtgcg gtcctcatca tgctgggcct
     1621 gcccgcctgc gtcctcttgc tgctgctgca gtgcaggcag aaggacccgg atccaatggc
     1681 ctcctccgag gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt
     1741 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1801 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     1861 ccctcagttc cagtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     1921 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1981 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2041 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     2101 ctgggaggcc tccaccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2161 gatgaggctg aagctgaagg acggcggcca ctacgacgcc gaggtcaaga ccacctacat
     2221 ggccaagaag cccgtgcagc tgcccggcgc ctacaagacc gacatcaagc tggacatcac
     2281 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgcgccgagg gccgccactc
     2341 caccggcgcc taagcggccg ctcgactatg ggcaccaaag aacctgtaaa cgttatcttt
     2401 ttaaattgaa tgtgcacaaa taaaagtttg gaaaagaaaa aaaaaaaaaa aaaaaaaaaa
     2461 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaat
     2521 acgcatcata cgctgtggcg tacctcgagt ctagagggcc cttcgaacaa aaactcatct
     2581 cagaagagga tctgaatatg cataccggtc atcatcacca tcaccattga gtttaaaccc
     2641 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2701 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2761 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2821 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2881 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     2941 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3001 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3061 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     3121 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3181 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3241 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     3301 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3361 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta
     3421 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3541 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3601 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3661 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3721 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3781 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3841 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3901 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3961 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4021 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4081 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4141 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4201 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4261 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4321 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4381 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4441 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4501 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4561 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4621 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4681 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4861 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4921 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4981 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5041 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5101 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5161 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5221 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     5521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5821 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5941 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6001 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
