LOCUS       sequence-417477-        8231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GLG1 fused with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  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 8231)
  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: GLG1-EGFP
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      4164..4173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4170..4172,4173..4175,4176..4889)
                     /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(4215..4236)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4476..4495)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4823..4844
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5012..5133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5049..5068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5103..5122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5140..5595)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5277..5298)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5489..5508
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5622..5726
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5728..6085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5728..5919
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5751..5771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5936..6071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6120..6914
                     /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             6120..6914
                     /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(6174..6193)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6784..6803
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7102..7121)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7146..7193
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7522..8110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8011..8030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctcga gatggcggcg tgtggacgtg tacggaggat gttccgcttg tcggcggcgc
      661 tgcatctgct gctgctattc gcggccgggg ccgagaaact ccccggccag ggcgtccaca
      721 gccagggcca gggtcccggg gccaactttg tgtccttcgt agggcaggcc ggaggcggcg
      781 gcccggcggg tcagcagctg ccccagctgc ctcagtcatc gcagcttcag cagcaacagc
      841 agcagcagca acagcaacag cagcctcagc cgccgcagcc gcctttcccg gcgggtgggc
      901 ctccggcccg gcggggagga gcgggggctg gtgggggctg gaagctggcg gaggaagagt
      961 cctgcaggga ggacgtgacc cgcgtgtgcc ctaagcacac ctggagcaac aacctggcgg
     1021 tgctcgagtg cctgcaggat gtgagggagc ctgaaaatga aatttcttca gactgcaatc
     1081 atttgttgtg gaattataag ctgaacctaa ctacagatcc caaatttgaa tctgtggcca
     1141 gagaggtttg caaatctact ataacagaga ttaaagaatg tgctgatgaa ccggttggaa
     1201 aaggttacat ggtttcctgc ttggtggatc accgaggcaa catcactgag tatcagtgtc
     1261 accagtacat taccaagatg acggccatca tttttagtga ttaccgttta atctgtggct
     1321 tcatggatga ctgcaaaaat gacatcaaca ttctgaaatg tggcagtatt cggcttggag
     1381 aaaaggatgc acattcacaa ggtgaggtgg tatcatgctt ggagaaaggc ctggtgaaag
     1441 aagcagaaga aagagaaccc aagattcaag tttctgaact ctgcaagaaa gccattctcc
     1501 gggtggctga gctgtcatcg gatgactttc acttagaccg gcatttatat tttgcttgcc
     1561 gagatgatcg ggagcgtttt tgtgaaaata cacaagctgg tgagggcaga gtgtataagt
     1621 gcctctttaa ccataaattt gaagaatcca tgagtgaaaa gtgtcgagaa gcacttacaa
     1681 cccgccaaaa gctgattgcc caggattata aagtcagtta ttcattggcc aaatcctgta
     1741 aaagtgactt gaagaaatac cggtgcaatg tggaaaacct tccgcgatcg cgtgaagcca
     1801 ggctctccta cttgttaatg tgcctggagt cagctgtaca cagagggcga caagtcagca
     1861 gtgagtgcca gggggagatg ctggattacc gacgcatgtt gatggaagac ttttctctga
     1921 gccctgagat catcctaagc tgtcgggggg agattgaaca ccattgttcc ggattacatc
     1981 gaaaagggcg gaccctacac tgtctgatga aagtagttcg aggggagaag gggaaccttg
     2041 gaatgaactg ccagcaggcg cttcaaacac tgattcagga gactgaccct ggtgcagatt
     2101 accgcattga tcgagctttg aatgaagctt gtgaatctgt aatccagaca gcctgcaaac
     2161 atataagatc tggagaccca atgatcttgt cgtgcctgat ggaacattta tacacagaga
     2221 agatggtaga agactgtgaa caccgtctct tagagctgca gtatttcatc tcccgggatt
     2281 ggaagctgga ccctgtcctg taccgcaagt gccagggaga cgcttctcgt ctttgccaca
     2341 cccacggttg gaatgagacc agtgaattta tgcctcaggg agctgtgttc tcttgtttat
     2401 acagacacgc ctaccgcact gaggaacagg gaaggaggct ctcacgggag tgccgagctg
     2461 aagtccaaag gatcctacac cagcgtgcca tggatgtcaa gctggatcct gccctccagg
     2521 ataagtgcct gattgatctg ggaaaatggt gcagtgagaa aacagagact ggacaggagc
     2581 tggagtgcct tcaggaccat ctggatgact tggtggtgga gtgtagagat atagttggca
     2641 acctcactga gttagaatca gaggatattc aaatagaagc cttgctgatg agagcctgtg
     2701 agcccataat tcagaacttc tgccacgatg tggcagataa ccagatagac tctggggacc
     2761 tgatggagtg tctgatacag aacaaacacc agaaggacat gaacgagaag tgtgccatcg
     2821 gagttaccca cttccagctg gtgcagatga aggattttcg gttttcttac aagtttaaaa
     2881 tggcctgcaa ggaggacgtg ttgaagcttt gcccaaacat aaaaaagaag gtggacgtgg
     2941 tgatctgcct gagcacgacc gtgcgcaatg acactctgca ggaagccaag gagcacaggg
     3001 tgtccctgaa gtgccgcagg cagctccgtg tggaggagct ggagatgacg gaggacatcc
     3061 gcttggagcc agatctatac gaagcctgca agagtgacat caaaaacttc tgttccgctg
     3121 tgcaatatgg caacgctcag attatcgaat gtctgaaaga aaacaagaag cagctaagca
     3181 cccgctgcca ccaaaaagta tttaagctgc aggagacaga gatgatggac ccagagctag
     3241 actacaccct catgagggtc tgtaagcaga tgataaagag gttctgtccg gaagcagatt
     3301 ctaaaaccat gttgcagtgc ttgaagcaaa ataaaaacag tgaattgatg gatcccaaat
     3361 gcaaacagat gataaccaag cgccagatca cccagaacac agattaccgc ttaaacccca
     3421 tgttaagaaa agcctgtaaa gctgacattc ctaaattctg tcacggtatc ctgactaagg
     3481 ccaaggatga ttcagaatta gaaggacaag tcatctcttg cctgaagctg agatatgctg
     3541 accagcgcct gtcttcagac tgtgaagacc agatccgaat cattatccag gagtccgccc
     3601 tggactaccg cctggatcct cagctccagc tgcactgctc agacgagatc tccagtctat
     3661 gtgctgaaga agcagcagcc caagagcaga caggtcaggt ggaggagtgc ctcaaggtca
     3721 acctgctcaa gatcaaaaca gaattgtgta aaaaggaagt gctaaacatg ctgaaggaaa
     3781 gcaaagcaga catctttgtt gacccggtac ttcatactgc ttgtgccctg gacattaaac
     3841 accactgcgc agccatcacc cctggccgcg ggcgtcaaat gtcctgtctc atggaagcac
     3901 tggaggataa gcgggtgagg ttacagcccg agtgcaaaaa gcgcctcaat gaccggattg
     3961 agatgtggag ttacgcagca aaggtggccc cagcagatgg cttctctgat cttgccatgc
     4021 aagtaatgac gtctccatct aagaactaca ttctctctgt gatcagtggg agcatctgta
     4081 tattgttcct gattggcctg atgtgtggac ggatcaccaa gcgagtgaca cgagagctca
     4141 aggacagggc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     4201 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4261 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4321 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4381 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4441 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4501 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4561 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4621 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4681 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4741 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag
     4801 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4861 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     4921 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4981 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5101 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     5161 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5221 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5281 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5341 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5401 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5461 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5521 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5581 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5641 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5701 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5761 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5821 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5881 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5941 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6001 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6061 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     6121 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6181 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6241 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6301 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6361 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6421 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6481 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6541 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6601 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     6661 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6721 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6781 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6841 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6901 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6961 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7021 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     7081 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     7141 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     7201 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     7261 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7321 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7381 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7441 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7501 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7561 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7621 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7681 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7741 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7801 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7861 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7921 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7981 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8041 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8101 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8161 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     8221 gcattagtta t
//
