LOCUS       sequence-133883-        5436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 N5-carboxyaminoimidazole 
            ribonucleotide mutase with inserted Val2 (AaPurE1-V2ins); 
            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 5436)
  AUTHORS   Kappock
  TITLE     Additional purEK associated constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5436)
  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            "Unpublished"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: pJK552
FEATURES             Location/Qualifiers
     source          1..5436
                     /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)."
     promoter        713..731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    732..756
                     /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             802..810
                     /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             2021..2065
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2103..2121)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2117..2164
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2335..3126)
                     /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(3127..3218)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3266..4004)
                     /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(4214..5296)
                     /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     5258..5277
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5297..5374)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5420..5436,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggtgagcg aaaccgcccc cctgccctct gcttccagtg ctttagaaga
      121 taaagcagcc tctgcccccg ttgtgggcat tatcatgggc agccagtcag actgggaaac
      181 catgcgccat gcagacgcat tgctgacaga gctggaaatt ccgcatgaaa ccctcattgt
      241 ttcagcacat cgtacgccag accggttggc ggattatgcg cgtacggcag ccgaacgcgg
      301 cctgaatgtg attattgctg gggccggtgg agctgcacat ctgccgggta tgtgtgcggc
      361 atggacacgc ctgcccgtgc tgggtgttcc agtagaatct cgcgctctta aaggcatgga
      421 cagcctgctt tccattgtgc agatgcccgg cggcgtgcct gtgggcactt tggctattgg
      481 ggcatccggc gccaaaaatg ccgccttgct ggccgcctcc attcttgcac tgtataaccc
      541 ggcccttgcc gcacgccttg aaacatggcg cgcattgcag acggcctctg tccccaactc
      601 tcccattacc gaagataaat gagaattcga gctccgtcga caagcttgcg gccgcataat
      661 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
      721 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
      781 taagaaggag atatacatat gacctcctta cattctccgc ttaagcccaa tgccgttatt
      841 ggcattgtgg ggggtggcca gcttggccgc atgtctgctg ttgctgccgc gcgccttggt
      901 tttcgcacgc atattcttac gccagaggcc aacggcccgg cggctcaggt ctctcacgcc
      961 gtaacatgcg gcaagtatga tgatccgaca gcactggatg cctttgcccg cgctgttgat
     1021 gttgttacct ttgaatttga aaacatcagc gcagatgcac tggaccgcct agccaaacac
     1081 tgccctgttc atccttcggg gcatattctg cgcatcagtc aggaccgatt ggcagaaaaa
     1141 actttttttg cttctgcggg tattccactc gcgccatggc atcctgttac cgatgtcaaa
     1201 acgctacacg aagccgcaga aaaagtgggt ttgccgctga ttctcaaaac cactcgcaat
     1261 ggatatgatg gcaaaggcca gcgccgtgtg catcacatcc atgatctgga agaggctttc
     1321 aacgctctgg caccacatcc tctggttgca gaaggtatgg tggattttgc ttgcgagatt
     1381 agtgttatgg tggcgcgtgg tgcagatggc acggtaaggt gctttgaccc ggcgctgaac
     1441 cggcactgga acggtattct ggatctaaca ttagcccccg cccccattga cccagatatt
     1501 gccacgcagg ccataaaact agccaccacc attgcagaaa agctggagct ggttggtatt
     1561 ctgggcgttg aaatgtttgt tgattgcacc ggcaacctgc tggtgaatga aatggccccg
     1621 cgcccccata attctggcca ttggaccatg aatgcctgcc cgcaggacca atttgacatg
     1681 catatccgtg cagttgcggg gctgcctttg ccgcacgctg tgcgccattc tgatgcaatc
     1741 atgaaaaatc tgttcgggcc ggaagatatg gcgctgctgc cgcagattat ggccacgcag
     1801 ggctttattc cgcatctata tggcaaggca gaagccagag tgggccgtaa aatggggcat
     1861 gtaaatattc tgttccccta cggtgcgctg ccaggcaatt tgggtattca ggatgccttg
     1921 ggcccattgg ccaccaaaac ggccgctgtg gaagatgaag aatctgccag ttaagaattc
     1981 gagctccgtc gacaagcttg cggccgcact cgagtctggt aaagaaaccg ctgctgcgaa
     2041 atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc
     2101 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2161 tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     2221 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     2281 cgggtcatcg tggccggatc ttgcggcccc tcggcttgaa cgaattgtta gacattattt
     2341 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     2401 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     2461 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     2521 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     2581 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     2641 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     2701 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     2761 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     2821 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     2881 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     2941 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     3001 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     3061 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     3121 cctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3181 cggatacata tttgaatgta tttagaaaaa taaacaaata gctagctcac tcggtcgcta
     3241 cgctccgggc gtgagactgc ggcgggcgct gcggacacat acaaagttac ccacagattc
     3301 cgtggataag caggggacta acatgtgagg caaaacagca gggccgcgcc ggtggcgttt
     3361 ttccataggc tccgccctcc tgccagagtt cacataaaca gacgcttttc cggtgcatct
     3421 gtgggagccg tgaggctcaa ccatgaatct gacagtacgg gcgaaacccg acaggactta
     3481 aagatcccca ccgtttccgg cgggtcgctc cctcttgcgc tctcctgttc cgaccctgcc
     3541 gtttaccgga tacctgttcc gcctttctcc cttacgggaa gtgtggcgct ttctcatagc
     3601 tcacacactg gtatctcggc tcggtgtagg tcgttcgctc caagctgggc tgtaagcaag
     3661 aactccccgt tcagcccgac tgctgcgcct tatccggtaa ctgttcactt gagtccaacc
     3721 cggaaaagca cggtaaaacg ccactggcag cagccattgg taactgggag ttcgcagagg
     3781 atttgtttag ctaaacacgc ggttgctctt gaagtgtgcg ccaaagtccg gctacactgg
     3841 aaggacagat ttggttgctg tgctctgcga aagccagtta ccacggttaa gcagttcccc
     3901 aactgactta accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa
     3961 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc
     4021 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     4081 ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     4141 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     4201 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4261 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4321 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4381 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4441 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     4501 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4561 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4621 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4681 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     4741 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     4801 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     4861 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     4921 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     4981 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5041 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     5101 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5161 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5221 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5281 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5341 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5401 gactcctgca ttaggaaatt aatacgactc actata
//
