LOCUS       sequence-141966-        4482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAL374 plasmid carrying the rfp (T) sgRNA targeting the template 
            strand of rfp, SpecR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4482)
  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 4482)
  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-rfp_T
FEATURES             Location/Qualifiers
     source          1..4482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            2111..2220
                     /note="incP origin of transfer"
     protein_bind    2952..2966
                     /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             3086..3877
                     /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"
     misc_RNA        4254..4329
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ggtgaggtta tggcggaggg ttgcgaggtc taggagaaca gaggaagtca tgctttgaag
       61 catataagct gccctgcccc tcaaggtttt cttcaagtga ggttttatct aactgcctaa
      121 cggcagggga accgtatatt gcttacggta tgagacccct taaacgtccg gatagtcacc
      181 gctcttcttt agctccgcga catgcctagc aaccgtggcg cgagagactc ctacctctgc
      241 ccctatttca gcccacgtgg gaactgtccc tgtctggaaa tactgatcgt tcaccatttg
      301 gctaatacgg gacttcgtag atcgtccttg agcctttttc ttacggtgcg tcttttcaag
      361 cttcgacctt tgtgcttgcg catatttgcc ctcggggtct gttttccagc gttgtgcggc
      421 tttttgtccg cctctgcgtc ccatcgtggc caaggctttc cgctcgctgc tggtggcttt
      481 acctggtgcg ttagagccgc tgtaggtctc gctcttggat tgggcgacat acccgcgcac
      541 gcgccttgcc atggtttggc ggtcgcgcat gggtggcatc tcgttgtcgc ggcctgcacc
      601 gccgtgggtg tgtgcgacgt tgtaggcgtg ctcataggcg tcgatgattg ctgcgtctgt
      661 caggcgttgg ccttgctggc gcaagcggtg gccagtctta agcgcatgtc taaaggctgt
      721 ttcgtcgcgt gctgcggttc cttggacaat ccagagcaca cgcacaccgt cgataagttc
      781 cgggtcatac tggtcgagac caccggcgat ttccgcgtct acgtcctggg cgagtgcttt
      841 gaatgcttgg gcttcttcac ggcgggtctt gaccgcgttg ataagttcgc ggccagagct
      901 gaattgctgg cgtggggtgg ggttgaactg gtcgtgtcct gccatatccc ttacctgctt
      961 tatcaagtct ccaaggcgca tcacccggtt gtgctgccta taccaacgat aagcggtagg
     1021 ggctttgcct gtgtagaacg ggttgcggct aaagcggtgg gaaaagtgcg ggtcatggtc
     1081 taaaagctca cccagcacac gcgtggttgc tgcaagaagc ttcatctgcg cagatttacc
     1141 gttacggtca gcgtagacag ggtcaataag ccatatgaac tgggctttgc cgttagttgg
     1201 gttaataccc acccaggctg gcccgacgct atgagtaatc agtgagcgca ccacgtcgcg
     1261 gacgtacggg tttaagtctg cggggtcacc gcctgcggta cctacttggt caacgtctac
     1321 gaccaggacg gcggcgtact gcttggtggt gagcatggcg tactcgcacc gtcctaaagc
     1381 atcagtctcg aagcgataca tacgcggcga gttcgtgccg tcagcgttgc gtcgataggc
     1441 ctttttaaag tctcgtgtga ctgaaccgtg gagtacatcg cggcctagat gatcgcgtaa
     1501 aaggtcgcgg tcactggcag atgctggggt gttgtccagt ccaccacggt cgcgctcgac
     1561 gcgggtaggt gttttagtgt gcgcattctg cgcatgagtc tgtaaactca tgaccgtgat
     1621 ttctcccagg tgtgtgctgg gtgataagcg aaagtcatcg ggttgccgcc cggtggcttt
     1681 cttcgttttt cattgtcttt ccctgactct aaatgacacc ggtgttattt actagccatg
     1741 acacgcgaaa aatatgcctt ttacctgcgg ttacaattcg cccttgctct agagatcgtg
     1801 gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc
     1861 aggcggccca ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg
     1921 cccgtgattt tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc
     1981 gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt
     2041 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     2101 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     2161 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     2221 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     2281 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     2341 tatgcagcgg aaaagcgctg cttccctgct gttttgtgga atatctaccg actggaaaca
     2401 ggcaaatgca ggaaattact gaactgaggg gacaggcgag agacgatgcc aaagagctac
     2461 accgacgagc tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg gcgtgatgag
     2521 gctgcggttg cgttcctggc ggtgagggcg gatgtcgagg cggcgttagc gtccggctat
     2581 gcgctcgtca ccatttggga gcacatgcgg gaaacgggga aggtcaagtt ctcctacgag
     2641 acgttccgct cgcacgccag gcggcacatc aaggccaagc ccgccgatgt gcccgcaccg
     2701 caggccaagg ctgcggaacc cgcgccggca cccaagacgc cggagccacg gcggccgaag
     2761 caggggggca aggctgaaaa gccggccccc gctgcggccc cgaccggctt caccttcaac
     2821 ccaacaccgg acaaaaagga tcaattcgcc cttgcaacgc gtgctgagcg acataaatac
     2881 cactggcggt gatactgagc acatcagcag gacgcactga ccaccatgaa ggtgacgctc
     2941 ttaaaaatta agccctgaag aagggcagca ttcaaagcag aaggctttgg ggtgtgtgat
     3001 acgaaacgaa gcattggccg taagtgcgat tccggattag ctgccaatga agtactagat
     3061 ccacggttag cggaggaggc cccccatgag ggaagcggtg atcgccgaag tatcgactca
     3121 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     3181 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     3241 tacggtgacc gtaaggcttg atgaaacaac gcggcaagct ttgatcaacg accttttgga
     3301 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     3361 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     3421 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     3481 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     3541 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     3601 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     3661 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     3721 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     3781 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     3841 cgtgaaaggc gagatcacca aggtagtcgg caaataattc gaaacgcgtt gcaagggcga
     3901 attggaaggg cgaattgtaa ccgcgtttca gctggcacga caggtttccc gactggaaag
     3961 cgggcagtga gcgcaacgca attaatgtaa gttagcgcga attgatctgg tttgacagct
     4021 tatcatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     4081 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     4141 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     4201 gttgacaatt aatcaggctc gtataatgaa gttcgtatgg aaggttccgt taagttttag
     4261 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     4321 agtcggtgct ttttatggaa ttggatccgt ggaagacatc tgtcgaagac accttgtggt
     4381 cgaagacacc ttgtctcttc ccagaggcgt ggtcgcatac taatttgtaa acggccatga
     4441 cgaaatagtc gtacggaaca aggagccact agtctagaat tc
//
