LOCUS       sequence-322812-        5653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses uvsY for bacterial expression and affinity purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  AUTHORS   Patchsung M, Homchan A, Aphicho K, Suraritdechachai S, Wanitchanon 
            T, Pattama A, Sappakhaw K, Meesawat P, Wongsatit T, Athipanyasilp A,
            Jantarug K, Athipanyasilp N, Buahom J, Visanpattanasin S, 
            Niljianskul N, Chaiyen P, Tinikul R, Wichukchinda N, Mahasirimongkol
            S, Sirijatuphat R, Angkasekwinai N, Crone MA, Freemont PS, Joung J, 
            Ladha A, Abudayyeh O, Gootenberg J, Zhang F, Chewapreecha C, 
            Chanarat S, Horthongkham N, Pakotiprapha D, Uttamapinant C
  TITLE     A Multiplexed Cas13-Based Assay with Point-of-Care Attributes for 
            Simultaneous COVID-19 Diagnosis and Variant Surveillance.
  JOURNAL   CRISPR J. 2022 Nov 11. doi: 10.1089/crispr.2022.0048.
  PUBMED    36367987
REFERENCE   2  (bases 1 to 5653)
  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: "CRISPR J. 
            2022 Nov 11. doi: 10.1089/crispr.2022.0048."
COMMENT     Sequence Label: pET28a-MH6-uvsY
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(646..1728)
                     /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     1690..1709
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1729..1806)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2012..2031
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2115..2134
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2115..2133
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2134..2158
                     /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)."
     promoter        join(2167..2172,2173..2190,2191..2196)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             2216..2238
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2270..2287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2702..2720)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2716..2763
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2800..3255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2887..2906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3097..3118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3347..4162)
                     /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(3347..4162)
                     /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     4070..4089
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4154..4173
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4284..4872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4773..4792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5026..5043
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5058..5197
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5123..5142)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5283..5305
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5299..5490)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
       61 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
      121 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
      181 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
      241 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
      301 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
      361 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
      421 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
      481 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
      541 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
      601 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
      661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      721 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
      781 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
      841 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
      901 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
      961 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     1021 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     1081 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     1141 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     1201 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     1261 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1321 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     1381 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     1441 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     1501 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     1561 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     1621 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     1681 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1741 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     1801 ggtgtccggg atctcgacgc tctcccttat gcggctcctg cattaggaag cagcccagta
     1861 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     1921 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     1981 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     2041 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     2101 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     2161 tctagattta cactttatgc ttccggctcg tatgtttcta gagaattcaa ataattttgt
     2221 ttaactttaa gaaggagatt taaatatgaa aatcgaagaa ggtaaaggtc accatcacca
     2281 tcaccacatg cgtctggaag atctgcaaga ggaactgaaa aaagatgtgt ttatcgacag
     2341 caccaaactg cagtatgaag cagcaaataa tgtgatgctg tatagcaaat ggctgaacaa
     2401 acatagcagc atcaaaaaag aaatgctgcg tattgaggcc cagaaaaaag ttgcactgaa
     2461 agcacgtctg gattattaca gcggtcgtgg tgatggtgat gaatttagca tggatcgtta
     2521 tgagaaaagc gagatgaaaa ccgttctgag cgcagataaa gatgttctga aagttgatac
     2581 cagcctgcag tattggggta ttctgctgga tttttgtagc ggtgcactgg atgcaattaa
     2641 aagccgtggt tttgccatta aacatatcca ggacatgcgt gcatttgaag ccggtaaata
     2701 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2761 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     2821 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2881 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2941 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3001 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3061 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3121 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3181 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3241 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3301 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     3361 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     3421 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     3481 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     3541 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3601 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3661 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     3721 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     3781 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     3841 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     3901 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     3961 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     4021 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     4081 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     4141 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     4201 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4261 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4321 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4381 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4441 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4501 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4561 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4621 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4681 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4741 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4801 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4861 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4921 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4981 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5041 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5101 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     5161 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     5221 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5281 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     5341 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     5401 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5461 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     5521 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     5581 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     5641 ggaccagaga aaa
//
