LOCUS       sequence-133893-        4144 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Produces Acetobacter aceti 1023 N5-carboxyaminoimidazole 
            ribonucleotide mutase with N-terminal His6 tag (H6AaPurE1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4144)
  AUTHORS   Kappock
  TITLE     Additional purEK associated constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4144)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pJK554
FEATURES             Location/Qualifiers
     source          1..4144
                     /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"
     CDS             110..127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             729..773
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(811..829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      825..872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1043..1834)
                     /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(1835..1926)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1974..2712)
                     /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(2922..4004)
                     /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     3966..3985
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4005..4082)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4128..4144,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4128..4144,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg
      121 cggcagccat atgatgagcg aaaccgcccc cctgccctct gcttccagtg ctttagaaga
      181 taaagcagcc tctgcccccg ttgtgggcat tatcatgggc agccagtcag actgggaaac
      241 catgcgccat gcagacgcat tgctgacaga gctggaaatt ccgcatgaaa ccctcattgt
      301 ttcagcacat cgtacgccag accggttggc ggattatgcg cgtacggcag ccgaacgcgg
      361 cctgaatgtg attattgctg gggccggtgg agctgcacat ctgccgggta tgtgtgcggc
      421 atggacacgc ctgcccgtgc tgggtgttcc agtagaatct cgcgctctta aaggcatgga
      481 cagcctgctt tccattgtgc agatgcccgg cggcgtgcct gtgggcactt tggctattgg
      541 ggcatccggc gccaaaaatg ccgccttgct ggccgcctcc attcttgcac tgtataaccc
      601 ggcccttgcc gcacgccttg aaacatggcg cgcattgcag acggcctctg tccccaactc
      661 tcccattacc gaagataaat gagaattcga gctccgtcga caagcttgcg gccgcactcg
      721 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
      781 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
      841 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
      901 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
      961 tttaacgacc ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc
     1021 ggcttgaacg aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt
     1081 agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa
     1141 gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
     1201 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
     1261 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata
     1321 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
     1381 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
     1441 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc
     1501 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
     1561 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
     1621 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
     1681 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
     1741 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
     1801 gagtcgatac ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa
     1861 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1921 aacaaatagc tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc
     1981 ggacacatac aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca
     2041 aaacagcagg gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca
     2101 cataaacaga cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga
     2161 cagtacgggc gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc
     2221 tcttgcgctc tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct
     2281 tacgggaagt gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc
     2341 gttcgctcca agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta
     2401 tccggtaact gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca
     2461 gccattggta actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga
     2521 agtgtgcgcc aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa
     2581 gccagttacc acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc
     2641 caggtggttt tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa
     2701 gatcctttga tcttttctac tgaaccgctc tagatttcag tgcaatttat ctcttcaaat
     2761 gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc
     2821 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     2881 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     2941 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3001 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     3061 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     3121 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     3181 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
     3241 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     3301 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     3361 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     3421 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     3481 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     3541 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     3601 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     3661 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     3721 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     3781 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     3841 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     3901 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     3961 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     4021 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     4081 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac
     4141 tata
//
