LOCUS       sequence-198314-        6948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Ran Q69L mutant in mammalian cells (CMV promoter). Also 
            suitable for in vitro transcription (T7 promoter) for injecting mRNA
            into oocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6948)
  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 6948)
  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-Ran-Q69L-mRFP1-polyA
FEATURES             Location/Qualifiers
     source          1..6948
                     /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"
     CDS             1562..2239
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1562..2239
                     /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(1692..1710)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1960..1979
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2154..2177
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2451..2480
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2496..2513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2536..2553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2542..2766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2812..3240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2899..2918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3109..3130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3249..3269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3254..3584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3435..3570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3497..3516
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3651..4445
                     /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(3705..3724)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4315..4334
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4621..4742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4658..4677)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4712..4731
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4791..4807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4791..4807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4804..4826)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4815..4831
                     /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(4839..4845,4846..4863,4864..4869))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4884..4905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5022..5039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5193..5781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5273..5292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5952..6743,6744..6812))
                     /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     6575..6594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6813..6917)
                     /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 taagcttatg gccgcccagg gagagccgca ggtccagttc aagctcgtcc tggtgggcga
      961 cggcggcacc gggaagacaa ccttcgtgaa gcgccacttg acgggcgagt ttgagaagaa
     1021 gtatgtagcc accctgggcg tggaggtgca cccgctcgtc ttccatacca acagaggacc
     1081 catcaagttc aacgtgtggg acacggccgg cctggagaag ttcgggggcc tgcgcgatgg
     1141 ctactacatc caagcccagt gtgccattat aatgtttgat gtaacctcaa gagttactta
     1201 caagaatgta cctaactggc atagagatct ggtacgagtg tgtgaaaaca tccccattgt
     1261 attgtgtggc aacaaagtgg atattaaaga caggaaagtg aaggcaaaat ctattgtctt
     1321 ccaccggaag aagaatcttc agtactatga catttctgcc aaaagtaact acaactttga
     1381 aaagcctttc ctctggcttg ccagaaagct cattggagat cctaacttgg agtttgttgc
     1441 catgcctgct cttgccccac ctgaggtggt catggaccca gctttggcag cacagtacga
     1501 gcatgattta gaggttgctc agacgactgc tctcccagat gaggatgatg acctggatcc
     1561 aatggcctcc tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg
     1621 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1681 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     1741 cctgtcccct cagttccagt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     1801 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1861 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     1921 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     1981 catgggctgg gaggcctcca ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2041 gatcaagatg aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac
     2101 ctacatggcc aagaagcccg tgcagctgcc cggcgcctac aagaccgaca tcaagctgga
     2161 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg
     2221 ccactccacc ggcgcctaag cggccgctcg actatgggca ccaaagaacc tgtaaacgtt
     2281 atctttttaa attgaatgtg cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa
     2341 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2401 aaaaatacgc atcatacgct gtggcgtacc tcgagtctag agggcccttc gaacaaaaac
     2461 tcatctcaga agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt
     2521 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2581 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2641 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2701 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2761 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2821 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2881 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2941 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     3001 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3061 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3121 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3181 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3241 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca
     3301 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3421 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3481 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3541 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3601 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3661 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3721 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3781 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3841 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3901 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3961 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4021 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4081 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4141 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4201 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4261 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4321 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4381 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4441 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     4501 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4561 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4621 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4681 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4741 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4801 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4861 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4921 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4981 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5041 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5101 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5161 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5221 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5281 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5341 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5401 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5461 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5521 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5581 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5641 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5701 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5821 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5881 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5941 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6001 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6061 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6121 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6181 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6241 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6301 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6361 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6421 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6481 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6541 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6601 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6661 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6721 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6781 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6841 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6901 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
