LOCUS       sequence-311067-        6451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for COVID-SARS2 NSP10 fused to GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6451)
  AUTHORS   Gileadi Lab
  TITLE     Bacterial co-expression vector for Covid-SARS2 NSP16 and NSP10
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6451)
  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: 
            "Unpublished"
COMMENT     Sequence Label: COVID-SARS2 NSP10
FEATURES             Location/Qualifiers
     source          1..6451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..1083)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1045..1064
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1084..1161)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1367..1386
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1470..1489
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1470..1488
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1489..1513
                     /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)."
     CDS             1557..2210
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             2217..2240
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2673..2693
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2694..2711
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2721..2735
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             2785..2802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2855..2873)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2869..2916
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2953..3408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3040..3059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3250..3271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3500..4315)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3500..4315)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4223..4242
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4307..4326
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4437..5025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4926..4945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5179..5196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5211..5350
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5276..5295)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5436..5458
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5452..5643)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
       61 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
      121 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
      181 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
      241 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
      301 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
      361 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
      421 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
      481 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
      541 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
      601 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
      661 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
      721 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
      781 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
      841 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
      901 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
      961 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     1021 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     1081 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     1141 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     1201 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     1261 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1321 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1381 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1441 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     1501 ggataacaat tcccctctag aaataatttt gtttaacctt aagaaggaga tatactatgt
     1561 ccccgatcct gggttactgg aaaatcaaag gtctggtcca gccgacgcgg ctgttgcttg
     1621 aatacttaga ggagaaatat gaggaacatc tgtatgaacg tgatgaaggc gacaaatggc
     1681 gtaataaaaa attcgagtta gggttagagt tccccaattt gccgtattat atcgacgggg
     1741 acgtgaaatt gactcagagc atggccatca ttcggtacat cgcagataag cataatatgt
     1801 taggtggctg ccctaaagaa cgggcagaaa tctctatgct ggaaggtgcc gtgctcgata
     1861 ttcgttacgg ggtatcccgc attgcctata gtaaagattt tgaaaccctg aaagtagact
     1921 ttctgtctaa actccccgaa atgcttaaaa tgtttgagga tcgcctgtgc cacaaaactt
     1981 atctgaacgg ggatcacgtg acccatccgg attttatgct ctatgatgcg ttagacgtgg
     2041 tgctttacat ggatccgatg tgtttagatg cgtttccgaa actggtgtgc ttcaaaaaac
     2101 ggattgaagc tatcccacaa atcgacaagt atttaaaaag cagcaaatat atcgcctggc
     2161 ccttacaggg ctggcaagct acgttcggtg gcggcgatca tccaccgaag tcggatctgg
     2221 aggtgctgtt tcaaggcccg ctgcgtagcg cgggaaatgc caccgaagtc ccagcaaatt
     2281 ctaccgtact gtcattttgt gcatttgccg tggacgcggc gaaggcctat aaggattatc
     2341 tggcatcggg tggtcagccg attaccaatt gtgtcaaaat gctgtgcacg catacgggca
     2401 cagggcaagc aatcacagtc acgccggaag ccaacatgga tcaggagtct tttggtggcg
     2461 cgagctgctg cctgtactgc cgttgtcaca ttgatcatcc gaacccgaaa ggtttctgtg
     2521 atttgaaagg taaatacgta caaatcccaa cgacctgtgc caacgatccg gtgggcttta
     2581 ccctgaaaaa cacggtctgc actgtgtgtg gcatgtggaa aggttacggc tgctcttgtg
     2641 atcagctgcg tgaaccgatg ttacaatgag cagagaacct ctacttccaa tcgcaccatc
     2701 atcaccacca ttgatacaag gatgacgacg ataagtgagg atccgaattc gagctccgtc
     2761 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
     2821 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2881 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     2941 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3001 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3061 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3121 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3181 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3241 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3301 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3361 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     3421 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3481 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     3541 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     3601 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     3661 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     3721 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     3781 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     3841 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     3901 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     3961 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     4021 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     4081 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     4141 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     4201 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     4261 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     4321 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     4381 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4441 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4501 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4561 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4621 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4681 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4741 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4801 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4861 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4921 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4981 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5041 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5101 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5161 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     5221 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     5281 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     5341 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5401 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5461 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     5521 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     5581 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     5641 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     5701 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     5761 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     5821 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     5881 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     5941 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     6001 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     6061 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     6121 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     6181 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     6241 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     6301 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     6361 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     6421 agtgagctaa cttacattaa ttgcgttgcg c
//
