LOCUS       sequence-262454-        7578 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express Nup37-EGFP 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 7578)
  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 7578)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Nup37-EGFP-polyA
FEATURES             Location/Qualifiers
     source          1..7578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             227..256
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             272..289
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(312..329)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    318..542
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      588..1016
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(675..694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     885..906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1025..1045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1030..1359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1210..1345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1272..1291
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1426..2220
                     /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(1480..1499)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2090..2109
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2396..2517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2433..2452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2487..2506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2566..2582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2566..2582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2579..2601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2590..2606
                     /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(2614..2620,2621..2638,2639..2644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2659..2680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2797..2814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2968..3556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3048..3067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3727..4518,4519..4587))
                     /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     4350..4369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4588..4692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4767..4786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4958..5337
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5338..5541
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5492..5512
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5586..5605
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5586..5604
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6628..6637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6634..6636,6637..6639,6640..7353)
                     /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(6634..6636,6637..6639,6640..7350)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6679..6700)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6940..6959)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7287..7308
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
       61 aaaaaaaaaa aanaaaaaaa annaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      181 atacgcatca tacgctgtgg cgtacctcga gtctagaggg cccttcgaac aaaaactcat
      241 ctcagaagag gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttaaac
      301 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
      361 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
      421 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
      481 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
      541 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
      601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      661 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
      721 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
      781 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
      841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
      901 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
      961 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1021 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1261 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     1321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     1381 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     1441 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     1501 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     1561 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     1621 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     1681 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     1741 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     1801 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     1861 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     1921 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     1981 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2041 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2101 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     2161 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     2221 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     2281 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     2341 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     2401 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2461 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2521 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     2581 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2641 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2701 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2761 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2821 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     4741 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4801 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     4861 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     4921 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
     4981 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     5041 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     5101 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     5161 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     5221 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     5281 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     5341 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     5401 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     5461 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     5521 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
     5581 gaaattaata cgactcacta tagggagacc caagctggct agttaagctt cgaattcatg
     5641 aagcaagatg ctacaagaaa tgctgcgtac accgtggatt gcgaagatta tgtgcacgtg
     5701 gtagaattta atccctttga gagtggagat tcaggaaacc tgattgcata tggcggcagt
     5761 aattatgtgg ttgttggcat gtgtacgttt caggaggaag aaacagacat cgaagggatt
     5821 cagtataaaa cactacgaac atttcaccat ggagtccgag ttgacggcat tgcctggagc
     5881 ccagagacca aacttgactc gctgcctcct gtaatcaaat tttgtacttc agctgctgat
     5941 ttgaagatta gattatttac ttcagatctt caagataaaa atgaatataa ggttttggag
     6001 ggccactcag acttcattaa tgatctggtg ttccatccca aagaaggcca ggagctcgca
     6061 agcgtgagtg acgaccacac ctgcaggatt tggaacttgg aaggaaagca gacagctcat
     6121 tttcttctgc attctcctgg aatgagtgta tgctggcatc ccgaggaaac tttcaagcta
     6181 atggttgcag agaagaatgg aacaatccgg ttttatgatc taatggctca gcaggcaatt
     6241 ttatctcttc agtcagagca gacaccctta atgtcagcac actggtgctt aaaaaacacc
     6301 ttcaaagttg gtgctgtggc aggaaatgat tggattattt gggatatcac tcggtccagt
     6361 tatcctcaag aaacaagacc tgttcacatg gaccgagcac acttatttag gtggtcggcc
     6421 attagtgaaa acttgtttgc aaccactgga tatcctggca aaatggccag tcagtttcag
     6481 attcatcatt tagggcactc tcagcccgtc ctcattggtt ctgtagctgt cggttctggt
     6541 ctttcctggc atagaacact tccactgtgt gcagttggag gcgaccacaa gctcttgttc
     6601 tgggtgacag aaatggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     6661 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     6721 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     6781 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     6841 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     6901 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     6961 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     7021 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     7081 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     7141 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     7201 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     7261 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     7321 atcactctcg gcatggacga gctgtacaag taaagcggcc gctcgactat gggcaccaaa
     7381 gaacctgtaa acgttatctt tttaaattga atgtgcacaa ataaaagttt ggaaaagaaa
     7441 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     7501 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     7561 aaaaaaaaaa aaaaaaaa
//
