LOCUS       sequence-178122-        6247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hQPRTase harbouring R161G mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6247)
  AUTHORS   Liu H, Woznica K, Catton G, Crawford A, Botting N, Naismith JH
  TITLE     Structural and kinetic characterization of quinolinate 
            phosphoribosyltransferase (hQPRTase) from homo sapiens.
  JOURNAL   J Mol Biol. 2007 Oct 26;373(3):755-63. Epub 2007 Aug 24.
  PUBMED    17868694
REFERENCE   2  (bases 1 to 6247)
  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 Mol 
            Biol."; date: "2007-10-26"; volume: "373(3)"; pages: "755-63. Epub 
            2007 Aug 24"
COMMENT     Sequence Label: pEHIAhqprtR161G
FEATURES             Location/Qualifiers
     source          1..6247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1101..1121)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1143..1160)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1180..1202
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1217..1241
                     /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)."
     primer_bind     complement(1241..1260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1242..1260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1348..1367)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1573..1650
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1651..2733
                     /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     complement(1670..1689)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3542..3733
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3727..3749)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3835..3977
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3890..3909
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3992..4009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4163..4751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4243..4262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4862..4881)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4873..5688
                     /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             4873..5688
                     /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     complement(4946..4965)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5781..6236)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5918..5939)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6130..6149
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccctagtgg attttgggca ctggagccac ctctttggca
      241 aacagcttga gggagaaatc aagggctggg accgcctggg tcagcatccc catggagatg
      301 acgtctatgt gcggcccgca gaactggggg aggttgtcca gggtgatgcc cccactggct
      361 tccacagcca cactcgggaa ctgggccttc agcgcggtgg ccgtggggtg cagctcctct
      421 ggcttgaagt tgtccagcag gacaaggtcg gcgccagcct cagctgcctg gacgacctcc
      481 tgcaggctgc tgcattccac ttccaccttc agagcgaagt cagccgcctg tctggccgcc
      541 cgcaccgcct tctccacgcc accggggggc accacatggt tatccttcaa catcaccagc
      601 cctcccaggt cgtacgcgtg cgaggcggcc ccgcccacca ggagcccata cttctccacc
      661 agccggaagc ctggcgtggt cttcctcgtg cctgccacgt gcccagtcca gccggccccc
      721 ctggcggcct ccactgcagc ggcggcagca ctggcaatgc cactgcagcg ggccagcgtg
      781 ttgagggcca cccgttcccc cagcagcagg cagtgggcag ggccccggac ctcggccact
      841 ctggccaccg gcaccagctt cgatccctcg gggaggaacc aggagacttg gcagttgagt
      901 tgggtaaata tggcatcgaa gaaaggctgc cctgccagta ccccagggga tttggcccac
      961 agcgccgcct gcgaggggcc tgccccgctg accaaggctg cgtagttgag ccctgggcag
     1021 tcctctcgga gccagctgtc caccagggct gccagggtga cgggcggcag cagcagcgcc
     1081 aggccttcag cgttcatggc gccctgaaaa tacaggtttt cggtcgttgg gatgtcgtaa
     1141 tcgtgatggt gatggtgatg gtagtacgac atatgtatat ctccttctta aagttaaaca
     1201 aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta
     1261 atttcgcggg atcgagatcg atctcgatcc tctacgccgg acgcatcgtg gccggcatca
     1321 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     1381 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     1441 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
     1501 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
     1561 gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
     1621 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     1681 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     1741 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     1801 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     1861 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     1921 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     1981 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     2041 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     2101 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     2161 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     2221 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     2281 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     2341 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     2401 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga catctcggta
     2461 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     2521 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     2581 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     2641 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2701 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     2761 cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2821 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2881 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     2941 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     3001 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     3061 cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     3121 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     3181 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     3241 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     3301 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     3361 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     3421 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     3481 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     3541 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     3601 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     3661 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
     3721 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3781 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     3841 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     3901 cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga
     3961 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4021 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4081 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4561 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4801 actcacgtta agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat
     4861 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat
     4921 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     4981 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     5041 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     5101 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     5161 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     5221 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     5281 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     5341 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     5401 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     5461 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     5521 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     5581 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     5641 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attaattcat
     5701 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5761 tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     5821 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5881 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     5941 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6001 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6061 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6121 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6181 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     6241 ttcgcca
//
