LOCUS       sequence-132599-        3954 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Produces Acetobacter aceti 1023 formylglycinamidine ribonucleotide 
            synthetase small subunit (AaPurS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3954)
  AUTHORS   Kappock
  TITLE     A multisubunit Gram-negative FGAM synthetase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3954)
  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            "Unpublished"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: pJK544
FEATURES             Location/Qualifiers
     source          1..3954
                     /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        387..405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    406..430
                     /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             539..583
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(621..639)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      635..682
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(853..1644)
                     /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(1645..1736)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1784..2522)
                     /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(2732..3814)
                     /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     3776..3795
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3815..3892)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(3938..3954,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcgaagg tacgtgtaac cgtcatgctg aaggacggtg ttctggaccc
      121 gcagggcaag gccatcagcc acgcattgca gacacttggt tttgaaggcg tgcacgatgt
      181 gcgcgtgggt aaggtgattg aactggatct gccgggtgcc aaagccgagg aagcccagaa
      241 aaaggccgat gctatggcgc gtgatctgct agccaacctg gtgattgaag attactctgt
      301 agaggtggtg gaatgaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
      361 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
      421 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatggcaga
      481 tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa
      541 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
      601 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      661 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
      721 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
      781 ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg
      841 aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
      901 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
      961 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     1021 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     1081 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     1141 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     1201 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     1261 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     1321 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     1381 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     1441 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     1501 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     1561 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     1621 ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca
     1681 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc
     1741 tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac
     1801 aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg
     1861 gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga
     1921 cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc
     1981 gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc
     2041 tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt
     2101 gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca
     2161 agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact
     2221 gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta
     2281 actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc
     2341 aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc
     2401 acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt
     2461 tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2521 tcttttctac tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     2581 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
     2641 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     2701 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2761 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2821 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     2881 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     2941 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     3001 ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     3061 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     3121 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     3181 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     3241 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     3301 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     3361 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     3421 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     3481 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     3541 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     3601 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     3661 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     3721 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     3781 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     3841 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     3901 cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac tata
//
