LOCUS       sequence-322669-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses uvsX for bacterial expression and affinity purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6391)
  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 6391)
  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-uvsX-H6
FEATURES             Location/Qualifiers
     source          1..6391
                     /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             3419..3436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3440..3458)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3454..3501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3538..3993
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3625..3644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3835..3856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4085..4900)
                     /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(4085..4900)
                     /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     4808..4827
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4892..4911
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5022..5610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5511..5530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5764..5781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5796..5935
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5861..5880)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6021..6043
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6037..6228)
                     /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 gcgactcctg 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 taaatatgag cgatctgaaa agccgtctga ttaaagcaag
     2281 caccagcaaa ctgaccgcag aactgacagc aagcaaattt ttcaacgaaa aagatgtggt
     2341 gcgcacgaaa attccgatga tgaatattgc actgagcggt gaaattaccg gtggtatgca
     2401 gagcggtctg ctgattctgg ctggtccgag caaaagcttt aaaagcaatt ttggtctgac
     2461 catggtgagc agctatatgc gtcagtatcc ggatgcagtt tgtctgtttt atgatagcga
     2521 atttggtatc acaccggcat atctgcgtag catgggtgtt gatccggaac gtgttattca
     2581 tacaccggtt cagagcctgg aacagctgcg tattgatatg gttaatcagc tggatgcaat
     2641 tgaacgtggt gaaaaagtgg tggtgtttat tgatagcctg ggtaatctgg caagcaaaaa
     2701 agaaaccgaa gatgccctga atgaaaaagt tgttagcgat atgacccgtg ccaaaaccat
     2761 gaaaagcctg tttcgtattg ttaccccgta tttcagcacc aaaaacattc cgtgtattgc
     2821 catcaaccat acctatgaaa cccaagagat gtttagcaaa accgttatgg gtggtggcac
     2881 cggtccgatg tatagcgcag ataccgtttt tatcattggc aaacgccaga ttaaagatgg
     2941 tagcgatctg cagggttatc agtttgttct gaatgttgaa aaaagccgca ccgtgaaaga
     3001 gaaaagcaaa ttctttatcg acgtgaaatt cgatggtggc attgatccgt atagtggtct
     3061 gctggatatg gcactggaac tgggttttgt tgttaaaccg aaaaatggtt ggtatgcccg
     3121 tgaatttctg gatgaagaaa ccggtgaaat gatccgtgaa gaaaaaagct ggcgtgcaaa
     3181 agataccaat tgtaccacct tttggggtcc gctgttcaaa catcagccgt ttcgtgatgc
     3241 cattaaacgt gcatatcagc tgggtgccat cgatagcaat gaaattgttg aagccgaagt
     3301 ggatgaactg atcaatagca aagtggaaaa attcaagagt ccggaatcca aaagcaaaag
     3361 cgcagcagat ctggaaaccg atctggaaca actgagcgat atggaagaat ttaacgaaca
     3421 ccatcaccat caccactaac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     3481 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3601 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3721 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3841 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3961 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4021 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     4081 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     4141 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4201 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4261 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4321 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     4381 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4441 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4501 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4561 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     4621 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4681 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4741 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     4801 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4861 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4921 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5101 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5161 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5221 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5281 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5341 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5401 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5461 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5521 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5581 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5641 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5701 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5761 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     5821 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     5881 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     5941 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6001 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6061 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     6121 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     6181 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     6241 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     6301 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     6361 ggcggtatgg atgcggcggg accagagaaa a
//
