LOCUS       sequence-131697-        4290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 N5-carboxyaminoimidazole 
            ribonucleotide mutase, Trp34>Phe/Trp165>Phe mutant (AaPurE1fwf).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4290)
  AUTHORS   Kappock
  TITLE     Single Trp variants of Acetobacter aceti PurE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4290)
  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            "Unpublished"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: pJK698
FEATURES             Location/Qualifiers
     source          1..4290
                     /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..768
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    787..803
                     /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)."
     promoter        complement(join(811..817,818..834,835..840,841..855))
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    819..835
                     /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(879..897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1452..1643
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1637..1659)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1745..1887
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1800..1819
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1902..1919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2073..2661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2153..2172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2832..3623,3624..3692))
                     /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     3455..3474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3693..3797)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3824..4279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3961..3982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4173..4192
                     /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 ggttatacag 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 tttcgaagtc tgactggctg
      661 cccatgataa tgcccacaac gggggcagag gctgctttat cttctaaagc actggaagca
      721 gagggcaggg gggcggtttc gctcatcata tgttaatttc tcctctttaa tgaattctgt
      781 gtgaaattgt tatccgctca caattgaatc tattataatt gttatccgct cacaaagcaa
      841 ataaattttt tatgatttct cgaggtgaag acgaaagggc ctcgtgatac gcctattttt
      901 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaga
      961 tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga
     1021 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     1081 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     1141 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     1201 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     1261 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     1321 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     1381 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     1441 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     1501 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     1561 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     1621 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     1681 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1741 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     1801 ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga
     1861 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     1921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3121 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     3181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
     3841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4261 gcgccgctac agggcgcgtc ccattcgcca
//
