LOCUS       sequence-418196-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the C terminal of SARS-CoV-2 NSP3 fused with mCherry tag 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  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 6310)
  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: NSP3C-mCherry
FEATURES             Location/Qualifiers
     source          1..6310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    621..645
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2259..2268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2265..2975
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2410..2428)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3097..3218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3134..3153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3188..3207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3225..3680)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3362..3383)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3574..3593
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3707..3811
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3813..4170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3813..4004
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3836..3856)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4021..4156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4205..4999
                     /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             4205..4999
                     /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(4259..4278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4869..4888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5187..5206)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5231..5278
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5607..6195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6096..6115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggactcaga tctcgagatg ccaactttgt acaaaaaagt tggcatgtgg ttcctgctgc
      661 tctccgtgtg cctgggttcc ctgatctact ctaccgccgc tctgggagtg ctgatgtcta
      721 acctgggtat gcctagctac tgcactggtt acagagaggg atacctgaac agcaccaacg
      781 tgaccatcgc cacctactgc accggcagca tcccatgctc cgtgtgcctg tctggactgg
      841 acagcctgga cacctacccc tccctggaaa ccatccagat caccatctct agcttcaagt
      901 gggacctgac cgccttcggt ctggtggctg agtggttcct ggcctacatc ctgttcacca
      961 ggttcttcta cgtgctgggc ctggccgcta tcatgcagct gttcttctcc tacttcgctg
     1021 tgcacttcat cagcaactcc tggctgatgt ggctgatcat caacctggtg cagatggctc
     1081 ccatctctgc catggtgcgc atgtacatct tcttcgccag cttctactac gtgtggaagt
     1141 cctacgtgca cgtggtggac ggttgcaact cctctacctg catgatgtgc tacaagcgca
     1201 acagggctac cagggtggag tgcaccacca tcgtgaacgg cgtgcgcagg tccttctacg
     1261 tgtacgccaa cggcggcaag ggattctgca agctgcacaa ctggaactgc gtgaactgcg
     1321 acaccttctg cgctggctcc accttcatct ctgacgaggt ggccagagac ctgagcctgc
     1381 agttcaagcg tccaatcaac cccaccgacc agagctccta catcgtggac tctgtgaccg
     1441 tgaagaacgg tagcatccac ctgtacttcg acaaggctgg ccagaagacc tacgagcgcc
     1501 actctctgag ccacttcgtg aacctggaca acctgagggc caacaacacc aagggcagcc
     1561 tgcctatcaa cgtgatcgtg ttcgacggaa agtccaagtg cgaggaatct agcgccaagt
     1621 ccgccagcgt gtactacagc cagctgatgt gccagccaat cctgctgctg gaccaggctc
     1681 tggtgtctga cgtgggagac agcgccgaag tggccgtgaa gatgttcgac gcctacgtga
     1741 acaccttctc ctctaccttc aacgtgccta tggagaagct gaagaccctg gtggctactg
     1801 ctgaggctga actggctaag aacgtgtccc tggacaacgt gctgagcacc ttcatctccg
     1861 ccgctaggca gggtttcgtg gactctgacg tggaaaccaa agacgtggtg gagtgcctga
     1921 agctgtctca ccagagcgac atcgaagtga ccggcgactc ctgcaacaac tacatgctga
     1981 cctacaacaa ggtggagaac atgaccccaa gagacctggg agcttgcatc gactgctccg
     2041 cccgtcacat caacgctcag gtggccaagt ctcacaacat cgccctgatc tggaacgtga
     2101 aggacttcat gagcctgtcc gagcagctgc gcaagcagat caggagcgcc gctaagaaga
     2161 acaacctgcc cttcaagctg acctgcgcta ccaccagaca ggtggtgaac gtggtgacca
     2221 ccaagatcgc cctgaagggc ggagcggatc caccggtcgc caccatggtg agcaagggcg
     2281 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     2341 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2401 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     2461 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2521 actacttgaa gctgtccttc cccgagggct tcaattggga gcgcgtgatg aacttcgagg
     2581 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2641 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag tgtcgtacca
     2701 tgggctggga ggcctccact gagcggatgt accccgagga cggcgccctg aagggcgaga
     2761 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2821 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcgacatc aagttggaca
     2881 tcctttccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2941 actccaccgg cggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc
     3001 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3061 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3121 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3181 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     3241 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3301 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3361 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3421 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3481 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3541 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3601 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3661 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3721 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3781 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3841 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3901 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4021 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4081 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4141 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4201 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4261 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4321 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4381 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4441 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4501 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4561 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4621 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4681 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4741 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4801 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4861 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4921 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4981 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5041 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5101 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5161 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5221 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5281 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5341 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5401 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6241 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6301 gccatgcatt
//
