LOCUS       sequence-133877-        4280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 N5-carboxyaminoimidazole 
            ribonucleotide mutase with inserted Val2 (AaPurE1-V2ins).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4280)
  AUTHORS   Kappock
  TITLE     Additional purEK associated constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4280)
  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            "Unpublished"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: pJK550
FEATURES             Location/Qualifiers
     source          1..4280
                     /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)."
     CDS             865..909
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(947..965)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      961..1008
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1179..1970)
                     /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(1971..2062)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2110..2848)
                     /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(3058..4140)
                     /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     4102..4121
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4141..4218)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4264..4280,1..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 ggcagatctc aattggatat cggccggcca cgcgatcgct
      841 gacgtcggta ccctcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac
      901 atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa
      961 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     1021 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     1081 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg
     1141 gatcttgcgg cccctcggct tgaacgaatt gttagacatt atttgccgac taccttggtg
     1201 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     1261 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     1321 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
     1381 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg
     1441 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
     1501 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
     1561 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
     1621 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
     1681 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
     1741 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
     1801 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
     1861 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
     1921 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt
     1981 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2041 tgtatttaga aaaataaaca aatagctagc tcactcggtc gctacgctcc gggcgtgaga
     2101 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
     2161 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
     2221 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
     2281 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
     2341 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
     2401 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
     2461 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
     2521 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
     2581 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
     2641 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
     2701 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
     2761 gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa
     2821 aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca
     2881 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     2941 atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     3001 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3121 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     3181 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     3241 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     3301 tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc
     3361 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     3421 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     3481 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     3541 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     3601 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     3661 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     3721 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     3781 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     3841 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     3901 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     3961 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     4021 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     4081 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     4141 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     4201 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     4261 aattaatacg actcactata
//
