LOCUS       sequence-410397-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Streptococcus pyogenes Inosine 5′-Monophosphate Dehydrogenase 
            (SpIMPDH, EC. 1.1.1.205). Contains His6 tag with TEV protease 
            cleavage site in pET-28b(+) plasmid for protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6756)
  AUTHORS   Glockzin K, Kostomiris D, Minnow YVT, Suthagar K, Clinch K, Gai S, 
            Buckler JN, Schramm VL, Tyler PC, Meek TD, Katzfuss A
  TITLE     Kinetic Characterization and Inhibition of Trypanosoma cruzi 
            Hypoxanthine-Guanine Phosphoribosyltransferases.
  JOURNAL   Biochemistry. 2022 Oct 4;61(19):2088-2105. doi: 
            10.1021/acs.biochem.2c00312. Epub 2022 Sep 15.
  PUBMED    36193631
REFERENCE   2  (bases 1 to 6756)
  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: 
            "10.1021/acs.biochem.2c00312"; journalName: "Biochemistry"; date: 
            "2022-10-4- 4"; volume: "61"; issue: "19"; pages: "2088-2105"
COMMENT     Sequence Label: SpIMPDH
FEATURES             Location/Qualifiers
     source          1..6756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     885..904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     988..1007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        988..1006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1007..1031
                     /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             1046..1068
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1081..1098
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1099..1119
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2386..2397
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2608..2625
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2678..2696)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2692..2739
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2776..3231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2863..2882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3073..3094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3323..4138)
                     /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(3323..4138)
                     /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     4046..4065
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4130..4149
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4260..4848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4749..4768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5002..5019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5034..5173
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5099..5118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5259..5281
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5275..5466)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6275..6756,1..601))
                     /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 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      961 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     1021 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc
     1081 catcaccacc accaccacga gaatctgtac ttccaaggca tgagcaactg ggacaccaaa
     1141 ttcctgaaaa agggctacac cttcgatgat gtgctgctga ttccggcgga gagccatgtg
     1201 ctgccgaacg aagttgacct gaagaccaaa ctggcggata acctgaccct gaacatcccg
     1261 atcattaccg cggcgatgga caccgtgacc ggtagcaaga tggcgattgc gattgcgcgt
     1321 gcgggtggcc tgggtgttat ccacaaaaac atgagcatta ccgagcaggc ggaggaagtg
     1381 cgtaaggtta aacgtagcga aaacggtgtg atcattgatc cgttctttct gaccccggag
     1441 cacaaggtta gcgaagcgga ggaactgatg caacgttacc gtatcagcgg cgtgccgatt
     1501 gttgaaaccc tggcgaaccg taaactggtg ggtatcatta ccaaccgtga catgcgtttc
     1561 atcagcgatt ataacgcgcc gattagcgag cacatgacca gcgaacacct ggtgaccgcg
     1621 gcggttggta ccgatctgga aaccgcggaa cgtatcctgc acgagcaccg tattgaaaag
     1681 ctgccgctgg ttgacaacag cggccgtctg agcggtctga tcaccattaa ggatatcgag
     1741 aaagtgattg aattcccgca cgcggcgaaa gacgagtttg gccgtctgct ggttgcggcg
     1801 gcggtgggtg ttaccagcga caccttcgag cgtgcggaag cgctgtttga agcgggcgcg
     1861 gacgcgatcg tgattgatac cgcgcacggc cacagcgcgg gtgttctgcg taagatcgcg
     1921 gagattcgtg cgcactttcc gaaccgtacc ctgatcgcgg gcaacattgc gaccgcggaa
     1981 ggtgcgcgtg cgctgtatga tgcgggtgtg gatgtggtta aagttggtat cggcccgggt
     2041 agcatttgca ccacccgtgt ggttgcgggc gtgggtgttc cgcaagtgac cgcgatctat
     2101 gatgcggcgg cggttgcgcg tgagtatggc aagaccatca ttgcggacgg tggcatcaag
     2161 tatagcggtg atattgttaa ggcgctggcg gcgggtggca acgcggttat gctgggcagc
     2221 atgttcgcgg gtaccgatga agcgccgggc gaaaccgaaa tctaccaggg tcgtaagttt
     2281 aaaacctatc gtggcatggg tagcattgcg gcgatgaaga aaggcagcag cgaccgttac
     2341 ttccaaggta gcgtgaacga ggcgaacaag ctggttccgg agggcatcga aggtcgtgtg
     2401 gcgtacaagg gtgcggcgag cgatatcgtt ttccaaatgc tgggtggcat tcgtagcggc
     2461 atgggttatg tgggcgcggg tgacatccag gagctgcacg aaaacgcgca atttgttgaa
     2521 atgagcggcg cgggtctgat tgaaagccac ccgcacgatg ttcagattac caacgaagcg
     2581 ccgaattaca gcgttcacta actcgagcac caccaccacc accactgaga tccggctgct
     2641 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2701 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2761 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2821 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2881 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2941 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3001 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3061 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3121 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3181 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     3241 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3301 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     3361 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     3421 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     3481 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     3541 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     3601 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     3661 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     3721 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     3781 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     3841 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     3901 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     3961 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4021 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     4081 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     4141 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     4201 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4261 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4321 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4381 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4441 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4501 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4561 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4621 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4681 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4741 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4801 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4861 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4921 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4981 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     5041 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     5101 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     5161 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5221 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5281 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5341 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5401 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5461 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     5521 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     5581 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     5641 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     5701 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5761 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5821 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5881 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     5941 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     6001 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     6061 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     6121 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     6181 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     6241 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6301 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6361 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     6421 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     6481 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     6541 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     6601 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     6661 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     6721 tcggctgaat ttgattgcga gtgagatatt tatgcc
//
