LOCUS       sequence-417564-        8069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses XPO6 fused with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  AUTHORS   Li J, Gui Q, Liang FX, Sall J, Zhang Q, Duan Y, Dhabaria A, Askenazi
            M, Ueberheide B, Stapleford KA, Pagano M
  TITLE     The REEP5/TRAM1 complex binds SARS-CoV-2 NSP3 and promotes virus 
            replication.
  JOURNAL   J Virol. 2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23.
  PUBMED    37768083
REFERENCE   2  (bases 1 to 8069)
  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"; journalName: "J Virol. 
            2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23."
COMMENT     Sequence Label: XPO6-EGFP
FEATURES             Location/Qualifiers
     source          1..8069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4013..4022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4019..4021,4022..4024,4025..4738)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4064..4085)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4325..4344)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4672..4693
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4861..4982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4898..4917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4952..4971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4989..5444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5126..5147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5338..5357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5471..5575
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5577..5934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5577..5768
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5600..5620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5785..5920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5969..6763
                     /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             5969..6763
                     /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(6023..6042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6633..6652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6951..6970)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6995..7042
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7371..7959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7860..7879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggac tcagatctcg agatggcatc tgaagaagcc tctctcaggg cattggaaag
      661 tctgatgaca gaattttttc acgattgtac aaccaatgaa agaaaacgtg agatagagga
      721 gcttcttaat aactttgccc agcaaatagg agcctggaga ttctgcctgt actttctctc
      781 cagcactagg aatgactatg taatgatgta cagtttaaca gtttttgaga atctgatcaa
      841 taaaatgtgg cttggggtcc catctcagga taagatggaa atccgtagct gtctgcccaa
      901 actccttttg gctcaccata aaaccttacc ttactttatc cggaacaagc tctgcaaagt
      961 tattgttgat attggacgtc aggattggcc catgttctac cacgactttt ttactaacat
     1021 tttacagttg atccagtccc ctgtgacaac cccccttggg ctgatcatgt tgaagacaac
     1081 ttcagaagag ctggcttgtc cccgtgagga cctcagtgtg gctcggaagg aggagttgcg
     1141 gaagctgcta ctggaccagg tgcagacagt gcttgggcta ctgacaggta tcttggagac
     1201 tgtctgggac aaacacagtg ttactgctgc cactccacca ccatccccga cctcaggaga
     1261 aagtggtgac ttactgagta acctgttgca gagtcccagt tcagccaaac tgttgaatca
     1321 gccaattccc atccttgatg tggagagtga gtatatctgt tccctggctt tggagtgcct
     1381 ggcccatctc ttcagttgga ttcctctgtc tgccagcatc accccatccc tccttaccac
     1441 catcttccac tttgcacgat ttggctgtga catccgggcc agaaagatgg cgtcagttaa
     1501 cggcagcagc cagaactgtg tctcgggtca ggagcgcggc cggctggggg tcctggccat
     1561 gtcctgcatc aatgaactca tgtccaagaa ctgtgtgcct atggaatttg aggagtattt
     1621 actgcgtatg ttccagcaga ctttctacct cctgcagaaa atcaccaagg ataacaatgc
     1681 ccacacagtg aagagcaggc tagaagagct cgatgagagc tatatcgaga agtttactga
     1741 ctttcttcgg ctctttgtga gtgttcacct aagaagaatc gagtcttatt cccagttccc
     1801 tgtggtggag tttttgacac ttttgttcaa gtacacattt catcagccta ctcatgaagg
     1861 ttacttctct tgtttggata tctggacgct gtttttggac tatctgacaa gtaaaattaa
     1921 aagtcgtctt ggagacaagg aagcagttct caacaggtac gaagatgccc tggtgctcct
     1981 gctcacagag gtgttgaatc gaatccagtt cagatacaac caagcccagc tggaggagtt
     2041 ggatgatgag actctggatg acgatcagca gacggagtgg cagcggtact tacggcagag
     2101 cttggaggtg gtggccaaag tgatggagct cctgcccacg cacgccttct ccacactgtt
     2161 ccctgttctt caggacaatt tagaagttta tttgggatta caacagttta tagtcacttc
     2221 agggtcagga cacaggttga acatcacggc ggagaacgac tgccggcggc tgcactgctc
     2281 cctgagagac ttgagctccc tgctgcaggc cgtgggccgc ctggccgagt actttatcgg
     2341 ggatgtgttt gctgcacggt tcaatgatgc cctcacagtc gtggaaaggt tggtcaaagt
     2401 cactctgtac ggatctcaga taaaattgta caacattgaa actgctgtgc catcagtatt
     2461 gaaacctgac ctcattgatg tgcatgctca gtccctggct gcgctgcagg cttactctca
     2521 ctggttagca cagtattgca gtgaagttca ccggcagaac acgcagcagt tcgtgacact
     2581 catctctact accatggatg caatcacacc tctaatcagc accaaggtcc aagacaagct
     2641 gctgctatct gcgtgccact tactggtctc actggccacc accgtgcggc ccgtctttct
     2701 gatcagcatc cctgcagtgc agaaagtatt caacagaatc actgatgcct ctgccctgcg
     2761 acttgtcgat aaggcccagg tgttggtgtg ccgagccctc tctaacatct tgctgcttcc
     2821 gtggccaaac cttccagaga atgagcagca gtggcccgtg cgctccatca accatgccag
     2881 cctcatctct gcactctccc gggactatcg caacctgaag cccagtgctg ttgccccaca
     2941 gagaaagatg ccactggatg acaccaaact gattatccac cagacactca gcgtcttaga
     3001 agatattgtg gagaatatct cgggggagtc caccaagtct cgacagattt gctaccagtc
     3061 gctgcaggaa tctgttcagg tctccctggc cctctttcca gcttttatcc atcagtcaga
     3121 tgtgactgat gagatgctga gcttcttcct cactctgttt cgaggcctta gagtacagat
     3181 gggtgtgcct ttcactgagc aaatcataca gactttcctc aacatgttta ccagagagca
     3241 gttagccgag agcatcctcc acgagggcag cacaggctgc cgggtggtgg agaagtttct
     3301 gaagatcctg caggtggtgg tccaggagcc aggccaggtg ttcaagccct tcctccccag
     3361 catcatcgcc ctgtgcatgg agcaagtgta tcccatcatt gccgagcgtc cctcccctga
     3421 tgtgaaggcc gagctgtttg agctcctttt ccggacgctc catcacaact ggaggtactt
     3481 cttcaagtcc accgtgctgg ccagtgtcca gagggggatc gctgaggagc agatggagaa
     3541 tgagccccag ttcagtgcca tcatgcaggc tttcggacag tcctttctcc agcccgacat
     3601 ccaccttttt aaacaaaatc tcttctactt ggagactctc aacaccaagc agaagctgta
     3661 ccacaagaag atcttccgga ctgccatgct gttccagttt gtgaacgtgc tgctccaggt
     3721 cctggtccac aagtcccatg atcttctgca ggaggagatt ggcatcgcca tctacaacat
     3781 ggcctcagtc gactttgatg gcttctttgc cgccttcctc ccagagttcc tgaccagctg
     3841 tgatggtgtg gatgccaacc agaaaagtgt gctggggcgg aatttcaaga tggatcggga
     3901 cctgccctca ttcacccaga atgtgcacag gctggtcaac gacctgcgct actacagact
     3961 ctgcaacgac agcctgcccc ctggcactgt gaagctcgcg gatccaccgg tcgccaccat
     4021 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4081 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4141 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4201 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     4261 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4321 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4381 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4441 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4501 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4561 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4621 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4681 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     4741 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4801 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4861 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4921 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4981 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5041 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     5101 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     5161 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     5221 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5281 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5341 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5401 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     5461 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5521 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     5581 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5701 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5761 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5821 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5881 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     5941 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     6001 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6061 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     6121 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     6181 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6241 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6301 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6361 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6421 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6481 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6541 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6601 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6661 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6721 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6781 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6841 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6901 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     6961 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     7021 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     7081 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     7141 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     7201 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     7261 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7321 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7381 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7441 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7501 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7561 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7621 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7681 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7741 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7801 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7861 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7921 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7981 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8041 gattctgtgg ataaccgtat taccgccat
//
