LOCUS       sequence-346372-        5873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Purification of AtNUDX23 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5873)
  AUTHORS   Sherwood AV, Rivera-Rangel LR, Ryberg LA, Larsen HS, Anker KM, Costa
            R, Vagbo CB, Jakljevic E, Pham LV, Fernandez-Antunez C, 
            Indrisiunaite G, Podolska-Charlery A, Grothen JER, Langvad NW, 
            Fossat N, Offersgaard A, Al-Chaer A, Nielsen L, Kusnierczyk A, 
            Solund C, Weis N, Gottwein JM, Holmbeck K, Bottaro S, Ramirez S, 
            Bukh J, Scheel TKH, Vinther J
  TITLE     Hepatitis C virus RNA is 5'-capped with flavin adenine dinucleotide.
  JOURNAL   Nature. 2023 Jul;619(7971):811-818. doi: 10.1038/s41586-023-06301-3.
            Epub 2023 Jul 5.
  PUBMED    37407817
REFERENCE   2  (bases 1 to 5873)
  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"; doi: 
            "10.1038/s41586-023-06301-3"; journalName: "Nature"; date: 
            "2023-07"; volume: "619"; issue: "7971"; pages: "811-818"
COMMENT     Sequence Label: pET30(a)-AtNUDX23
FEATURES             Location/Qualifiers
     source          1..5873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     973..992
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1012..1089)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1295..1314
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1402..1421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1402..1420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1421..1445
                     /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)."
     RBS             1460..1482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1493..1510
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2135..2152
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2205..2223)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2219..2266
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2303..2758
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2390..2409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2600..2621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2850..3665)
                     /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(2850..3665)
                     /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     3573..3592
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3657..3676
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3787..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4276..4295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4529..4546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4561..4700
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4626..4645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4786..4808
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4802..4993)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5802..5873,1..1011))
                     /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"
ORIGIN
        1 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
       61 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      121 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      181 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      241 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      301 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      361 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      421 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      481 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      541 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      601 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      661 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      721 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      781 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      841 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
      901 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
      961 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1021 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1081 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1141 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1201 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1261 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1321 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1381 tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     1441 attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca
     1501 tcaccaccac gtacaatcag ctggggatgt taggaagata aagttctgcc aatggtgcgg
     1561 tggcccgacc aagcacgaga ttccggatgg cgaggaaaaa ctgcgtgcga tctgcaccca
     1621 ctgcggcaag attgcgtacc agaacccgaa aatggtggtt ggttgcctga tcgagcacga
     1681 aggcaaggtg ctgctgtgca aacgtaacat tcaaccgagc catggtctgt ggaccctgcc
     1741 ggcgggttat ctggaagtgg gtgaaagcgc ggcgcaaggt gcgatgcgtg aaacctggga
     1801 ggaagcgggt gcgaccgtgg aagttatcag cccgttcgcg cagctggaca tcccgctgat
     1861 tggccaaacc tacgtgattt tcctggcgaa gctgaaaaac ctgcactttg cgccgggtcc
     1921 ggagagcctg gaatgccgtc tgtttgcgct ggacgagatc ccgttcgata gcctggcgtt
     1981 tagcagcatt tacgttaccc tgaacctgta tctggaagat ctgaagaaag gtaaactgaa
     2041 attccactac ggcaccatca acaagcgtcc gggtagcagc ccgagcgaca ttcgtgcgtt
     2101 tagcctggat tatcacctgc aaccgtaact cgagcaccac caccaccacc actgagatcc
     2161 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2221 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     2281 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2341 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2401 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2461 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2521 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2581 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2641 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2701 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     2761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2821 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     2881 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     2941 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     3001 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     3061 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     3121 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     3181 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     3241 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     3301 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     3361 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     3421 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3481 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     3541 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3601 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     3661 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     3721 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4501 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4561 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     4621 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     4681 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4741 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4801 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4861 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4921 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4981 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     5041 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     5101 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     5161 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     5221 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     5281 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     5341 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     5401 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     5461 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5521 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5581 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5641 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     5701 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     5761 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     5821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg aga
//
