LOCUS       sequence-177630-        7213 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To express human Nup93 in mammalian cells. It contains synonymous 
            mutations that are not targeted by shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7213)
  AUTHORS   Tie HC, Madugula V, Lu L
  TITLE     The development of a single molecule fluorescence standard and its 
            application in estimating the stoichiometry of the nuclear pore 
            complex.
  JOURNAL   Biochem Biophys Res Commun. 2016 Sep 30;478(4):1694-9. doi: 
            10.1016/j.bbrc.2016.09.005. Epub 2016 Sep 6.
  PUBMED    27613095
REFERENCE   2  (bases 1 to 7213)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbrc.2016.09.005"; journalName: "Biochem Biophys Res 
            Commun"; date: "2016-09-30- 30"; volume: "478"; issue: "4"; pages: 
            "1694-9"
COMMENT     Sequence Label: pEGFP-C1X-Nup93-mut
FEATURES             Location/Qualifiers
     source          1..7213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..601
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     552..572
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      640..649
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(646..648,649..651,652..1362)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(646..648,649..651,652..1362)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(691..712)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(952..971)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1299..1320
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4034..4155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4071..4090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4125..4144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4162..4617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4299..4320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4511..4530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4644..4748
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4750..5107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4750..4941
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4773..4793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4958..5093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5142..5936
                     /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             5142..5936
                     /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(5196..5215)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5806..5825
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6124..6143)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6168..6215
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6544..7132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7033..7052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      601 tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc
      661 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      721 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      781 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      841 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      901 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      961 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1021 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1081 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1141 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1201 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1261 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1321 accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga
     1381 cttcccggga tgacggctgt agcacaagga tctgcatctc caatggatac tgaggggttt
     1441 ggtgagctcc ttcagcaagc tgaacagctt gctgctgaga ctgagggcat ctcagagctt
     1501 ccccatgtgg aacggaactt acaggagatc cagcaggcgg gagagcgcct gcgttcccgt
     1561 accctaacac gcacgtccca ggagacggca gatgtcaagg cgtcagttct cctcgggtct
     1621 cggggacttg acatatccca catctcccag cgattggaga gtctgagtgc agccaccacc
     1681 tttgagcctc ttgagcctgt gaaggacact gacattcagg gcttcctgaa gaatgagaag
     1741 gacaatgccc tgctgtctgc catcgaagag tcccggaaga ggaccttcgg catggctgag
     1801 gagtaccatc gggagtcaat gttggttgag tgggagcaag tgaaacagcg aattctgcac
     1861 acactgctgg catcaggaga agacgccctt gactttactc aagaaagcga gccaagctac
     1921 atcagtgatg tgggaccccc tggtcgaagc tctctggata acatcgagat ggcctatgcg
     1981 cggcaaattt atatctataa tgagaaaatt gtaaatggac acctgcagcc taacctggtg
     2041 gacctttgtg cttccgtcgc agagctcgat gataagagca tttccgacat gtggaccatg
     2101 gtaaaacaaa tgacagacgt gttgttgaca ccggcaacgg atgccctgaa gaaccgcagc
     2161 agcgtggaag tgcgcatgga gtttgtcagg caggccttgg cgtaccttga gcagagttat
     2221 aagaattaca cccttgtgac tgtctttgga aatttgcatc aggcccagct gggcggggtg
     2281 cctgggactt accaattggt tcgaagtttc ctgaacatta aactgccagc tcccttgcct
     2341 ggactacagg atggagaggt ggaaggccat cctgtgtggg cgctaattta ctactgcatg
     2401 cgctgtggag acctgcttgc cgcttcacag gtagttaatc gagcccagca ccagctggga
     2461 gagtttaaaa cctggttcca ggagtacatg aacagcaagg acagaagatt gtccccagct
     2521 acggaaaaca agctccggct gcattaccgt agggccctca ggaacaatac agatccctac
     2581 aagcgggccg tgtactgtat cattggcaga tgtgacgtca ccgacaacca gagtgaagtg
     2641 gcggacaaaa ctgaggatta cctgtggctg aagttgaacc aagtgtgttt tgacgacgat
     2701 ggcaccagct ccccacaaga caggctcact ctctcacagt tccagaagca gttgttggaa
     2761 gactatggcg agtcccactt tacggtgaac cagcaaccct tcctctactt ccaagtcctg
     2821 ttcctgacag cgcagtttga agcagcagtt gcctttcttt tccgcatgga gcggctgcgc
     2881 tgccatgctg tccatgtagc actggtgctg tttgagctga agctgctttt aaagtcctct
     2941 ggacagaggg ctcagctcct cagccacgag cctggtgacc ctccttgctt gcggcggctg
     3001 aacttcgtgc ggctcctcat gctgtacacc cggaagtttg agtccacgga cccaagggag
     3061 gccctccagt acttctattt cctcagggat gagaaagata gtcaaggaga aaacatgttt
     3121 ctgcgctgtg tgagtgagct tgtgattgaa agccgagagt tcgatatgat tcttgggaaa
     3181 ctagagaatg acggaagtag aaagcctgga gtcatagata agtttactag tgatacgaaa
     3241 cccatcatta ataaagttgc ttctgtggca gaaaataaag gactgtttga agaggcagca
     3301 aagctgtatg accttgccaa gaatgctgac aaggtactgg agctgatgaa caaactgctg
     3361 agccctgtcg tcccccagat cagtgccccg caatccaaca aggagaggct gaagaacatg
     3421 gcactctcca ttgccgaacg gtatagggct caaggaataa gcgcaaataa atttgtggac
     3481 tccacgttct atcttctttt ggacttgatc accttttttg acgagtatca tagtggtcat
     3541 attgatagag cttttgatat cattgagcgc ttgaagctgg tgcccctgaa tcaggaaagt
     3601 gtggaagaga gagtggctgc cttcagaaat ttcagtgatg aaatcaggca caacctctca
     3661 gaagtgcttc ttgccaccat gaacatcttg ttcacacagt ttaagaggct caaggggaca
     3721 agtccatcct cgtcatccag gccccagcga gtcatcgagg accgcgactc tcaactccga
     3781 agtcaagccc gcactctgat tacctttgct ggaatgatac cataccgaac gtctggggac
     3841 accaatgcga ggctggtgca gatggaagtt ttaatgaact agctcgagga tccgagctcc
     3901 gcggagatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3961 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4021 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4081 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4141 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4621 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4681 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4741 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4861 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4921 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4981 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5041 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5101 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5161 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5221 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5281 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5341 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5401 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5461 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5521 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5581 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5641 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5701 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5761 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5821 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5881 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5941 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6001 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6061 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6121 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6181 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6241 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6301 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6361 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7201 tgcgttatcc cct
//
