LOCUS       sequence-131690-        4176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 N5-carboxyaminoimidazole 
            ribonucleotide mutase, Tyr154>Phe/Trp165>Phe mutant 
            (AaPurE1wwf-Y154F).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4176)
  AUTHORS   Kappock
  TITLE     Single Trp variants of Acetobacter aceti PurE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4176)
  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            "Unpublished"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: pJK663
FEATURES             Location/Qualifiers
     source          1..4176
                     /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             757..779
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(810..829)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(811..829)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1338..1529
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1523..1545)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1631..1773
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1686..1705
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1788..1805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1959..2547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2039..2058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2718..3509,3510..3578))
                     /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     3341..3360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3579..3683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3710..4165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3847..3868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4059..4078
                     /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 cgaattctca tttatcttcg gtaatgggag agttggggac agaggccgtc
      241 tgcaatgcgc gaaatgtttc aaggcgtgcg gcaagggccg ggttaaacag tgcaagaatg
      301 gaggcggcca gcaaggcggc atttttggcg ccggatgccc caatagccaa agtgcccaca
      361 ggcacgccgc cgggcatctg cacaatggaa agcaggctgt ccatgccttt aagagcgcga
      421 gattctactg gaacacccag cacgggcagg cgtgtccatg ccgcacacat acccggcaga
      481 tgtgcagctc caccggcccc agcaataatc acattcaggc cgcgttcggc tgccgtacgc
      541 gcataatccg ccaaccggtc tggcgtacga tgtgctgaaa caatgagggt ttcatgcgga
      601 atttccagct ctgtcagcaa tgcgtctgca tggcgcatgg tttcccagtc tgactggctg
      661 cccatgataa tgcccacaac gggggcagag gctgctttat cttctaaagc actggaagca
      721 gagggcaggg gggcggtttc gctcatcata tgtatatctc cttcttaaag ttaaacaaaa
      781 ttatttctag agggaaaccg ttgtggtctc cctatagtga gtcgtattaa tttcgcggga
      841 tcgagatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt
      901 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
      961 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     1021 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     1081 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     1141 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     1201 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     1261 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     1321 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     1381 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     1441 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     1501 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1561 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     1621 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     1681 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg
     1741 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     1801 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1861 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3001 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     3061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3301 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3661 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga aattgtaaac
     3721 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3781 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3841 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3901 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3961 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4021 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4081 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4141 cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
