LOCUS       sequence-133906-        3583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Franciscella tularensis N5-carboxyaminoimidazole 
            ribonucleotide mutase with N-terminal TEV protease-cleavable His6 
            tag (H6tFtPurE1), synthetic gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3583)
  AUTHORS   Kappock
  TITLE     Additional purEK associated constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3583)
  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            "Unpublished"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: pJK569
FEATURES             Location/Qualifiers
     source          1..3583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     445..464
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        445..463
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    464..488
                     /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             503..525
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             536..553
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             578..598
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1141..1158
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1211..1229)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1225..1272
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1362..1378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1375..1397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1386..1402
                     /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(1410..1416,1417..1434,1435..1440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1455..1476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1593..1610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1764..2352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1844..1863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2523..3314,3315..3383))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3146..3165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3384..3488)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3556..3574
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tgacccaatg aaattaatac gactcactat aggggaattg tgagcggata
      481 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgcacca
      541 tcatcatcat cattcaagcg gcgtggatct gggcaccgaa aatctgtact ttcagtctaa
      601 catgtcagtt caggttggcg tcattatggg cagcaaatct gattggagta ccatgaaaga
      661 atgctgtgat attctggaca acctgggcat cggttatgaa tgcgaagtgg ttagcgcgca
      721 tcgtacgccg gacaaaatgt tcgattacgc agaaacggcg aaagaacgcg gtctgaaagt
      781 catcatcgct ggcgcgggtg gtgctgcaca tttacctgga atggttgcgg caaaaactac
      841 tttaccagta ttaggtgtac cagttaaatc aagtacactt aatggccaag atagtttatt
      901 atcaatagta cagatgccag cgggtattcc agtagctact tttgcaattg gtatggcagg
      961 agcaaaaaac gcggcgcttt ttgctgcaag tatcttgcaa catacagata taaatattgc
     1021 aaaagcacta gctgagttta gagctgagca aacacgattt gttttggaaa atcctgatcc
     1081 tagggagcac taacggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag
     1141 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     1201 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1261 aggggttttt tgctgaaagg aggaactata tccggatgac ccaaatcgga tcccgggccc
     1321 gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1381 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1441 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1501 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1561 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1621 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1681 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1921 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2161 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2221 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2521 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2761 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2821 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2881 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2941 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3001 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3061 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3121 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3181 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3241 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3301 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3361 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3421 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3481 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     3541 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
