LOCUS       sequence-133874-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 N5-carboxyaminoimidazole 
            ribonucleotide synthetase, Val319>Phe mutant (AaPurK-V319F).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  AUTHORS   Kappock
  TITLE     Additional purEK associated constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4937)
  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            "Unpublished"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: pJK547
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /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             303..311
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1522..1566
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1604..1622)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1618..1665
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1836..2627)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2628..2719)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2767..3505)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3715..4797)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4759..4778
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4798..4875)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4921..4937,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
      301 tgacctcctt acattctccg cttaagccca atgccgttat tggcattgtg gggggtggcc
      361 agcttggccg catgtctgct gttgctgccg cgcgccttgg ttttcgcacg catattctta
      421 cgccagaggc caacggcccg gcggctcagg tctctcacgc cgtaacatgc ggcaagtatg
      481 atgatccgac agcactggat gcctttgccc gcgctgttga tgttgttacc tttgaatttg
      541 aaaacatcag cgcagatgca ctggaccgcc tagccaaaca ctgccctgtt catccttcgg
      601 ggcatattct gcgcatcagt caggaccgat tggcagaaaa aacttttttt gcttctgcgg
      661 gtattccact cgcgccatgg catcctgtta ccgatgtcaa aacgctacac gaagccgcag
      721 aaaaagtggg tttgccgctg attctcaaaa ccactcgcaa tggatatgat ggcaaaggcc
      781 agcgccgtgt gcatcacatc catgatctgg aagaggcttt caacgctctg gcaccacatc
      841 ctctggttgc agaaggtatg gtggattttg cttgcgagat tagtgttatg gtggcgcgtg
      901 gtgcagatgg cacggtaagg tgctttgacc cggcgctgaa ccggcactgg aacggtattc
      961 tggatctaac attagccccc gcccccattg acccagatat tgccacgcag gccataaaac
     1021 tagccaccac cattgcagaa aagctggagc tggttggtat tctgggcgtt gaaatgtttg
     1081 ttgattgcac cggcaacctg ctggtgaatg aaatggcccc gcgcccccat aattctggcc
     1141 attggaccat gaatgcctgc ccgcaggacc aatttgacat gcatatccgt gcagttgcgg
     1201 ggctgccttt gccgcacgct gtgcgccatt ctgatgcaat catgaaaaat ctgttcgggc
     1261 cggaagatat ggcgctgctg ccgcagatta tggccacgca gggctttatt ccgcatctat
     1321 atggcaaggc agaagccaga gtgggccgta aaatggggca tgtaaatatt ctgttcccct
     1381 acggtgcgct gccaggcaat ttgggtattc aggatgcctt gggcccattg gccaccaaaa
     1441 cggccgctgt ggaagatgaa gaatctgcca gttaagaatt cgagctccgt cgacaagctt
     1501 gcggccgcac tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg
     1561 gactcgtcta ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta
     1621 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca
     1681 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     1741 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcatc gtggccggat
     1801 cttgcggccc ctcggcttga acgaattgtt agacattatt tgccgactac cttggtgatc
     1861 tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
     1921 tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc
     1981 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
     2041 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
     2101 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
     2161 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
     2221 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
     2281 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
     2341 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
     2401 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
     2461 acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg
     2521 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
     2581 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatact cttccttttt
     2641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2701 atttagaaaa ataaacaaat agctagctca ctcggtcgct acgctccggg cgtgagactg
     2761 cggcgggcgc tgcggacaca tacaaagtta cccacagatt ccgtggataa gcaggggact
     2821 aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt tttccatagg ctccgccctc
     2881 ctgccagagt tcacataaac agacgctttt ccggtgcatc tgtgggagcc gtgaggctca
     2941 accatgaatc tgacagtacg ggcgaaaccc gacaggactt aaagatcccc accgtttccg
     3001 gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc
     3061 cgcctttctc ccttacggga agtgtggcgc tttctcatag ctcacacact ggtatctcgg
     3121 ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg ttcagcccga
     3181 ctgctgcgcc ttatccggta actgttcact tgagtccaac ccggaaaagc acggtaaaac
     3241 gccactggca gcagccattg gtaactggga gttcgcagag gatttgttta gctaaacacg
     3301 cggttgctct tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga tttggttgct
     3361 gtgctctgcg aaagccagtt accacggtta agcagttccc caactgactt aaccttcgat
     3421 caaaccacct ccccaggtgg ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa
     3481 aggatctcaa gaagatcctt tgatcttttc tactgaaccg ctctagattt cagtgcaatt
     3541 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg
     3601 ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3661 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     3721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3781 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     3841 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     3901 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     3961 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
     4021 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4081 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4141 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4201 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4261 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4321 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4381 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4441 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     4501 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     4561 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     4621 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     4681 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     4741 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     4801 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     4861 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat
     4921 taatacgact cactata
//
