LOCUS       sequence-131666-        4137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Treponema denticola 5-aminoimidazole ribonucleotide 
            carboxylase, Met15>Trp mutant (TdPurE2-M15W).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4137)
  AUTHORS   Kappock
  TITLE     Serine mutants of Treponema denticola PurE2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4137)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJK696
FEATURES             Location/Qualifiers
     source          1..4137
                     /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"
     RBS             718..740
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(771..790)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(772..790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1299..1490
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1484..1506)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1592..1734
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1647..1666
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1749..1766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1920..2508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2000..2019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2679..3470,3471..3539))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3302..3321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3540..3644)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3671..4126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3808..3829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4020..4039
                     /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 cgaattcgag ctccgtcgac aagaaactga aacgctgaat tcagcgtttc
      241 agtttcgaat cgtcttcgat aattttctgc gcattggatt ccatatagga tttgaccgaa
      301 tctgcgattt ctttatcgta cagtgaaaaa atacgagccg ccaggagtgc cgcatttttc
      361 ggttcaagaa ccagtgccgg actaatcccg ctcggcatcc gtaacgaaga ataaatgtcg
      421 gcgccggcga agctatcgct tggcggtggg caggcaatgg ttgcaccttt gacgaagcca
      481 tcaacaaagc ctgataaagc gttactacgc ccagcgatcg tgatgtacag tttaggacgg
      541 tccaaagctt catattcttt cagcatagac actacatgct ccgctgtttt atgcgcggag
      601 ccgatgcgaa tggcatactc aataccaaaa gttttcaatt cgctagcaat tttctcggca
      661 tgcccccaat ctgaactgga tcccataagg ataatcacca gcggacgcat atgtatatct
      721 ccttcttaaa gttaaacaaa attatttcta gagggaaacc gttgtggtct ccctatagtg
      781 agtcgtatta atttcgcggg atcgagatct cgggcagcgt tgggtcctgg ccacgggtgc
      841 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt
      901 agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg
      961 cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc
     1021 ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac
     1081 cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc
     1141 tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg
     1201 catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta
     1261 cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg
     1321 cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg
     1381 agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc
     1441 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1501 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1561 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     1621 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     1681 tatatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     1741 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1801 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1861 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1921 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1981 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2041 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2101 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2161 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2221 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2281 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2341 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2401 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2461 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2521 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2581 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2641 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2701 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2761 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2821 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2881 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2941 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     3001 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3061 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3121 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3181 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3241 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3301 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3361 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3421 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3481 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3541 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3601 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3661 gtgccacctg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3721 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3781 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3841 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3901 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3961 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4021 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4081 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//
