LOCUS       sequence-405024-        4484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAL374 plasmid carrying the pgi (NT) sgRNA, targeting the 
            non-template strand of pgi, SpecR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4484)
  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 4484)
  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-pgi_NT
FEATURES             Location/Qualifiers
     source          1..4484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..97
                     /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             217..1008
                     /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"
     promoter        join(1334..1338,1339..1354,1355..1361)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_RNA        1386..1461
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     oriT            3726..3835
                     /note="incP origin of transfer"
ORIGIN
        1 gacataaata ccactggcgg tgatactgag cacatcagca ggacgcactg accaccatga
       61 aggtgacgct cttaaaaatt aagccctgaa gaagggcagc attcaaagca gaaggctttg
      121 gggtgtgtga tacgaaacga agcattggcc gtaagtgcga ttccggatta gctgccaatg
      181 aagtactaga tccacggtta gcggaggagg ccccccatga gggaagcggt gatcgccgaa
      241 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
      301 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
      361 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcaagc tttgatcaac
      421 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
      481 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
      541 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
      601 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
      661 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
      721 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
      781 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
      841 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
      901 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
      961 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataatt cgaaacgcgt
     1021 tgcaagggcg aattggaagg gcgaattgta accgcgtttc agctggcacg acaggtttcc
     1081 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg aattgatctg
     1141 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
     1201 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
     1261 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
     1321 tgaaatgagc tgctgacaat taatcatcgg ctcgtataat gttgcctgga agtttgagta
     1381 atgatgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     1441 aaagtggcac cgagtcggtg cttttttatg gaattggatc cgtggaagac atctgtcgaa
     1501 gacaccttgt ggtcgaagac accttgtctc ttcccagagg cgtggtcgca tactaatttg
     1561 taaacggcca tgacgaaata gtcgtacgga acaaggagcc actagtctag aattcggtga
     1621 ggttatggcg gagggttgcg aggtctagga gaacagagga agtcatgctt tgaagcatat
     1681 aagctgccct gcccctcaag gttttcttca agtgaggttt tatctaactg cctaacggca
     1741 ggggaaccgt atattgctta cggtatgaga ccccttaaac gtccggatag tcaccgctct
     1801 tctttagctc cgcgacatgc ctagcaaccg tggcgcgaga gactcctacc tctgccccta
     1861 tttcagccca cgtgggaact gtccctgtct ggaaatactg atcgttcacc atttggctaa
     1921 tacgggactt cgtagatcgt ccttgagcct ttttcttacg gtgcgtcttt tcaagcttcg
     1981 acctttgtgc ttgcgcatat ttgccctcgg ggtctgtttt ccagcgttgt gcggcttttt
     2041 gtccgcctct gcgtcccatc gtggccaagg ctttccgctc gctgctggtg gctttacctg
     2101 gtgcgttaga gccgctgtag gtctcgctct tggattgggc gacatacccg cgcacgcgcc
     2161 ttgccatggt ttggcggtcg cgcatgggtg gcatctcgtt gtcgcggcct gcaccgccgt
     2221 gggtgtgtgc gacgttgtag gcgtgctcat aggcgtcgat gattgctgcg tctgtcaggc
     2281 gttggccttg ctggcgcaag cggtggccag tcttaagcgc atgtctaaag gctgtttcgt
     2341 cgcgtgctgc ggttccttgg acaatccaga gcacacgcac accgtcgata agttccgggt
     2401 catactggtc gagaccaccg gcgatttccg cgtctacgtc ctgggcgagt gctttgaatg
     2461 cttgggcttc ttcacggcgg gtcttgaccg cgttgataag ttcgcggcca gagctgaatt
     2521 gctggcgtgg ggtggggttg aactggtcgt gtcctgccat atcccttacc tgctttatca
     2581 agtctccaag gcgcatcacc cggttgtgct gcctatacca acgataagcg gtaggggctt
     2641 tgcctgtgta gaacgggttg cggctaaagc ggtgggaaaa gtgcgggtca tggtctaaaa
     2701 gctcacccag cacacgcgtg gttgctgcaa gaagcttcat ctgcgcagat ttaccgttac
     2761 ggtcagcgta gacagggtca ataagccata tgaactgggc tttgccgtta gttgggttaa
     2821 tacccaccca ggctggcccg acgctatgag taatcagtga gcgcaccacg tcgcggacgt
     2881 acgggtttaa gtctgcgggg tcaccgcctg cggtacctac ttggtcaacg tctacgacca
     2941 ggacggcggc gtactgcttg gtggtgagca tggcgtactc gcaccgtcct aaagcatcag
     3001 tctcgaagcg atacatacgc ggcgagttcg tgccgtcagc gttgcgtcga taggcctttt
     3061 taaagtctcg tgtgactgaa ccgtggagta catcgcggcc tagatgatcg cgtaaaaggt
     3121 cgcggtcact ggcagatgct ggggtgttgt ccagtccacc acggtcgcgc tcgacgcggg
     3181 taggtgtttt agtgtgcgca ttctgcgcat gagtctgtaa actcatgacc gtgatttctc
     3241 ccaggtgtgt gctgggtgat aagcgaaagt catcgggttg ccgcccggtg gctttcttcg
     3301 tttttcattg tctttccctg actctaaatg acaccggtgt tatttactag ccatgacacg
     3361 cgaaaaatat gccttttacc tgcggttaca attcgccctt gctctagaga tcgtggcatc
     3421 accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg
     3481 gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca cgacgcccgt
     3541 gattttgtag ccctggccga cggccagcag gtaggccgac aggctcatgc cggccgccgc
     3601 cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga taggtgggct
     3661 gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc
     3721 ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     3781 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     3841 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     3901 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     3961 agcggaaaag cgctgcttcc ctgctgtttt gtggaatatc taccgactgg aaacaggcaa
     4021 atgcaggaaa ttactgaact gaggggacag gcgagagacg atgccaaaga gctacaccga
     4081 cgagctggcc gagtgggttg aatcccgcgc ggccaagaag cgccggcgtg atgaggctgc
     4141 ggttgcgttc ctggcggtga gggcggatgt cgaggcggcg ttagcgtccg gctatgcgct
     4201 cgtcaccatt tgggagcaca tgcgggaaac ggggaaggtc aagttctcct acgagacgtt
     4261 ccgctcgcac gccaggcggc acatcaaggc caagcccgcc gatgtgcccg caccgcaggc
     4321 caaggctgcg gaacccgcgc cggcacccaa gacgccggag ccacggcggc cgaagcaggg
     4381 gggcaaggct gaaaagccgg cccccgctgc ggccccgacc ggcttcacct tcaacccaac
     4441 accggacaaa aaggatcaat tcgcccttgc aacgcgtgct gagc
//
