LOCUS       sequence-314169-        4481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAL374 plasmid carrying the pyk (NT) sgRNA targeting the 
            non-template strand of pyk, SpecR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4481)
  AUTHORS   Cleto S, Jensen JV, Wendisch VF, Lu TK
  TITLE     Corynebacterium glutamicum Metabolic Engineering with CRISPR 
            Interference (CRISPRi).
  JOURNAL   ACS Synth Biol. 2016 Feb 16.
  PUBMED    26829286
REFERENCE   2  (bases 1 to 4481)
  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: "ACS Synth 
            Biol. 2016 Feb 16."
COMMENT     Sequence Label: pAL-pyk_NT
FEATURES             Location/Qualifiers
     source          1..4481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..76
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     oriT            2341..2450
                     /note="incP origin of transfer"
     protein_bind    3182..3196
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     CDS             3316..4107
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRQALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
       61 ggcaccgagt cggtgctttt ttatggaatt ggatccgtgg aagacatctg tcgaagacac
      121 cttgtggtcg aagacacctt gtctcttccc agaggcgtgg tcgcatacta atttgtaaac
      181 ggccatgacg aaatagtcgt acggaacaag gagccactag tctagaattc ggtgaggtta
      241 tggcggaggg ttgcgaggtc taggagaaca gaggaagtca tgctttgaag catataagct
      301 gccctgcccc tcaaggtttt cttcaagtga ggttttatct aactgcctaa cggcagggga
      361 accgtatatt gcttacggta tgagacccct taaacgtccg gatagtcacc gctcttcttt
      421 agctccgcga catgcctagc aaccgtggcg cgagagactc ctacctctgc ccctatttca
      481 gcccacgtgg gaactgtccc tgtctggaaa tactgatcgt tcaccatttg gctaatacgg
      541 gacttcgtag atcgtccttg agcctttttc ttacggtgcg tcttttcaag cttcgacctt
      601 tgtgcttgcg catatttgcc ctcggggtct gttttccagc gttgtgcggc tttttgtccg
      661 cctctgcgtc ccatcgtggc caaggctttc cgctcgctgc tggtggcttt acctggtgcg
      721 ttagagccgc tgtaggtctc gctcttggat tgggcgacat acccgcgcac gcgccttgcc
      781 atggtttggc ggtcgcgcat gggtggcatc tcgttgtcgc ggcctgcacc gccgtgggtg
      841 tgtgcgacgt tgtaggcgtg ctcataggcg tcgatgattg ctgcgtctgt caggcgttgg
      901 ccttgctggc gcaagcggtg gccagtctta agcgcatgtc taaaggctgt ttcgtcgcgt
      961 gctgcggttc cttggacaat ccagagcaca cgcacaccgt cgataagttc cgggtcatac
     1021 tggtcgagac caccggcgat ttccgcgtct acgtcctggg cgagtgcttt gaatgcttgg
     1081 gcttcttcac ggcgggtctt gaccgcgttg ataagttcgc ggccagagct gaattgctgg
     1141 cgtggggtgg ggttgaactg gtcgtgtcct gccatatccc ttacctgctt tatcaagtct
     1201 ccaaggcgca tcacccggtt gtgctgccta taccaacgat aagcggtagg ggctttgcct
     1261 gtgtagaacg ggttgcggct aaagcggtgg gaaaagtgcg ggtcatggtc taaaagctca
     1321 cccagcacac gcgtggttgc tgcaagaagc ttcatctgcg cagatttacc gttacggtca
     1381 gcgtagacag ggtcaataag ccatatgaac tgggctttgc cgttagttgg gttaataccc
     1441 acccaggctg gcccgacgct atgagtaatc agtgagcgca ccacgtcgcg gacgtacggg
     1501 tttaagtctg cggggtcacc gcctgcggta cctacttggt caacgtctac gaccaggacg
     1561 gcggcgtact gcttggtggt gagcatggcg tactcgcacc gtcctaaagc atcagtctcg
     1621 aagcgataca tacgcggcga gttcgtgccg tcagcgttgc gtcgataggc ctttttaaag
     1681 tctcgtgtga ctgaaccgtg gagtacatcg cggcctagat gatcgcgtaa aaggtcgcgg
     1741 tcactggcag atgctggggt gttgtccagt ccaccacggt cgcgctcgac gcgggtaggt
     1801 gttttagtgt gcgcattctg cgcatgagtc tgtaaactca tgaccgtgat ttctcccagg
     1861 tgtgtgctgg gtgataagcg aaagtcatcg ggttgccgcc cggtggcttt cttcgttttt
     1921 cattgtcttt ccctgactct aaatgacacc ggtgttattt actagccatg acacgcgaaa
     1981 aatatgcctt ttacctgcgg ttacaattcg cccttgctct agagatcgtg gcatcaccga
     2041 accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca
     2101 ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt
     2161 tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc gccgccgcct
     2221 tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt gggctgccct
     2281 tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag
     2341 ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
     2401 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
     2461 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
     2521 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
     2581 aaaagcgctg cttccctgct gttttgtgga atatctaccg actggaaaca ggcaaatgca
     2641 ggaaattact gaactgaggg gacaggcgag agacgatgcc aaagagctac accgacgagc
     2701 tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg gcgtgatgag gctgcggttg
     2761 cgttcctggc ggtgagggcg gatgtcgagg cggcgttagc gtccggctat gcgctcgtca
     2821 ccatttggga gcacatgcgg gaaacgggga aggtcaagtt ctcctacgag acgttccgct
     2881 cgcacgccag gcggcacatc aaggccaagc ccgccgatgt gcccgcaccg caggccaagg
     2941 ctgcggaacc cgcgccggca cccaagacgc cggagccacg gcggccgaag caggggggca
     3001 aggctgaaaa gccggccccc gctgcggccc cgaccggctt caccttcaac ccaacaccgg
     3061 acaaaaagga tcaattcgcc cttgcaacgc gtgctgagcg acataaatac cactggcggt
     3121 gatactgagc acatcagcag gacgcactga ccaccatgaa ggtgacgctc ttaaaaatta
     3181 agccctgaag aagggcagca ttcaaagcag aaggctttgg ggtgtgtgat acgaaacgaa
     3241 gcattggccg taagtgcgat tccggattag ctgccaatga agtactagat ccacggttag
     3301 cggaggaggc cccccatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     3361 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     3421 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     3481 gtaaggcttg atgaaacaac gcggcaagct ttgatcaacg accttttgga aacttcggct
     3541 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     3601 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     3661 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     3721 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     3781 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     3841 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     3901 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     3961 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     4021 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     4081 gagatcacca aggtagtcgg caaataattc gaaacgcgtt gcaagggcga attggaaggg
     4141 cgaattgtaa ccgcgtttca gctggcacga caggtttccc gactggaaag cgggcagtga
     4201 gcgcaacgca attaatgtaa gttagcgcga attgatctgg tttgacagct tatcatcgac
     4261 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
     4321 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
     4381 ttttttgcgc cgacatcata acggttctgg caaatattct gaatgagctg tacaattaat
     4441 catcggctcg tataatgatt ccatctgcac tagccaccgc t
//
