LOCUS       sequence-142240-        5191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Escherichia coli N5-carboxyaminoimidazole ribonucleotide 
            mutase (EcPurE1-H45W) and N5-carboxyaminoimidazole ribonucleotide 
            synthetase (EcPurK).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Hoskins AA, Morar M, Kappock TJ, Mathews II, Zaugg JB, Barder TE, 
            Peng P, Okamoto A, Ealick SE, Stubbe J
  TITLE     N5-CAIR mutase: role of a CO2 binding site and substrate movement in
            catalysis.
  JOURNAL   Biochemistry. 2007 Mar 13;46(10):2842-55. Epub 2007 Feb 14.
  PUBMED    17298082
REFERENCE   2  (bases 1 to 5191)
  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: 
            "Biochemistry."; date: "2007-03-13"; volume: "46(10)"; pages: 
            "2842-55. Epub 2007 Feb 14"
COMMENT     Sequence Label: pJK048
FEATURES             Location/Qualifiers
     source          1..5191
                     /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             1772..1794
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1825..1844)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1826..1844)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2353..2544
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2538..2560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2646..2788
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2701..2720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2803..2820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2974..3562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3054..3073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3733..4524,4525..4593))
                     /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     4356..4375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4594..4698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4725..5180)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4862..4883)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5074..5093
                     /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 gcaagcttgc
      181 atgcctgcag ttaaccgaac ttactctgcg cccaaatcac gccgctggca tattccggcg
      241 gcagcagcgg gattaaggct tccagcgtcg cagtcagacg cgatgtgtcg ctgtcggtca
      301 aattcagatg ccccacttta cgccccggac ggacttcttt gtcgtaccag tgcagatgca
      361 ccagcggcag tttcagccag tcataattca catcgctacc aatcagattg atcatcaccg
      421 acggattatt caccactggt tgcggtaacg gcagatcggt aatcgcccgc agatgcagct
      481 caaactggct gatgctggca ccgttttgtg tccagtgacc gctgttatgc acacgcggtg
      541 ccagttcgtt gatcaacaga ccttgcgggg tgacaaaaca ctccatcgcc atcacgccca
      601 catagcccag ctcctgcata atcgccgaca gcatctcttc ggctcgcgcc tgctgctgtg
      661 cgttggcctg cggaaaagcg acgctggtgc gcaaaatacc gtcctgatgc aggttatgcg
      721 tcagcggata aaacacggtg ctgccatcaa agccgcgcgc gccaaccagc gacacttcac
      781 cagagaagtt aatgccctgc tcgacaatac attcgccgta acactctgcc ggtaactgtt
      841 cggtttcatt tgcgcgtaaa cgccattgac cgcgaccgtc ataaccacca gtgcgacgct
      901 taacaatcgc cagctcacct aaacgatcaa acaccgcagg ccactcgctg cgttcggcaa
      961 gtaactgcca cggtgcagtc ggcaggtgga gcttatcgaa aagctgcttc tgagtcagac
     1021 ggtcagcaat aatcgggaac acatcgcggt tcacaaaggc cggatggcgc gccagctcgc
     1081 gggttaatgc ggtttccggc cagcgttcta tctcagcggt aatcacgctt tgttgaaaag
     1141 gcaccgccgc cggttcagcg tccagcccga ctggccagac agcaatgcct aacggttcgc
     1201 ctgcctgacg cagcatacgg cctaactgcc cgttaccgag gacgcaaacc tgtttcatgc
     1261 cgcacctcgc gggtccgggt tttccagcac ttcgtcggtc tgggctttgc gccagtcatt
     1321 cagacgctgg tgcagttctt tatcatgagt cgcaagaatt tgtgctgcca gtaacgccgc
     1381 gtttgccgcg ccagctttac caatcgccag cgtacccacc ggaatgccgc gcggcatttg
     1441 tacgatggag tagaggctat cgacaccgct cagtgcggcg ctctgtactg gcacgcccag
     1501 caccggcacc agcgttttgg cggcaatcat gcctggcaga tgcgctgcgc cgcctgcgcc
     1561 cgcaataatc acctgataac cgttctcttc ggcgctttcg gcgaagctga acagtttatc
     1621 gggggtgcgc caagcagaaa ccacttcaac gtggtgcggg acattcagga tttcgaagat
     1681 ttcggcggcg aactgcatgg tagcccagtc gcttttggac cccatcacaa tggcgacacg
     1741 cgccggatta ttgcgggaag acatatgtat atctccttct taaagttaaa caaaattatt
     1801 tctagaggga aaccgttgtg gtctccctat agtgagtcgt attaatttcg cgggatcgag
     1861 atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     1921 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     1981 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     2041 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     2101 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     2161 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     2221 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     2281 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     2341 acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     2401 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     2461 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc
     2521 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2581 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2641 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     2701 cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa taccgcacag
     2761 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     2821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4021 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     4081 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4141 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4201 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4261 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4621 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4681 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa
     4741 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4801 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4861 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4921 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4981 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5041 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5101 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5161 tgcgccgcta cagggcgcgt cccattcgcc a
//
