LOCUS       sequence-438911-        3282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA expression vector - pBG42 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3282)
  AUTHORS   Roghair Stroud MN, Vang DX, Halverson LJ
  TITLE     Optimized CRISPR Interference System for Investigating Pseudomonas 
            alloputida Genes Involved in Rhizosphere Microbiome Assembly.
  JOURNAL   ACS Synth Biol. 2024 Aug 20. doi: 10.1021/acssynbio.4c00312.
  PUBMED    39163848
REFERENCE   2  (bases 1 to 3282)
  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. 2024 Aug 20. doi: 10.1021/acssynbio.4c00312."
COMMENT     Sequence Label: pSpyC
FEATURES             Location/Qualifiers
     source          1..3282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(56..142)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_RNA        249..324
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      326..369
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      462..556
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(677..696)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             688..1503
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1670..1778
                     /note="incP origin of transfer"
     CDS             complement(1789..2451)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2452..3223
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 gaagcccgga gagcaagccc gtaggggggg cgcgcccagc tgtctagggc ggcggatttg
       61 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
      121 actgagcctt tcgttttatt tgatgccttt aattaagccc attgacaagg ctctcgcggc
      181 caggtataat tgcacgacct aggccgcggc cgcgcgaatt cgagctcggt ctcatcacta
      241 gggtctctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      301 gaaaaagtgg caccgagtcg gtgctataaa acgaaaggcc cagtctttcg actgagcctt
      361 tcgttttata actaggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg
      421 cggccgcgtc gtgactggga aaaccctggc gactagtctt ggactcctgt tgatagatcc
      481 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
      541 ttttattggt gagaatccag gggtccccaa taattacgat ttaaatttgt gtctcaaaat
      601 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
      661 tacataaaca gtaatacaag gggtgttatg agccatattc agcgtgaaac gagctgtagc
      721 cgtccgcgtc tgaacagcaa catggatgcg gatctgtatg gctataaatg ggcgcgtgat
      781 aacgtgggtc agagcggcgc gaccatttat cgtctgtatg gcaaaccgga tgcgccggaa
      841 ctgtttctga aacatggcaa aggcagcgtg gcgaacgatg tgaccgatga aatggtgcgt
      901 ctgaactggc tgaccgaatt tatgccgctg ccgaccatta aacattttat tcgcaccccg
      961 gatgatgcgt ggctgctgac caccgcgatt ccgggcaaaa ccgcgtttca ggtgctggaa
     1021 gaatatccgg atagcggcga aaacattgtg gatgcgctgg ccgtgtttct gcgtcgtctg
     1081 catagcattc cggtgtgcaa ctgcccgttt aacagcgatc gtgtgtttcg tctggcccag
     1141 gcgcagagcc gtatgaacaa cggcctggtg gatgcgagcg attttgatga tgaacgtaac
     1201 ggctggccgg tggaacaggt gtggaaagaa atgcataaac tgctgccgtt tagcccggat
     1261 agcgtggtga cccacggcga ttttagcctg gataacctga ttttcgatga aggcaaactg
     1321 attggctgca ttgatgtggg ccgtgtgggc attgcggatc gttatcagga tctggccatt
     1381 ctgtggaact gcctgggcga atttagcccg agcctgcaaa aacgtctgtt tcagaaatat
     1441 ggcattgata atccggatat gaacaaactg caatttcatc tgatgctgga tgaatttttc
     1501 taataattaa ttggaccgcg gtccgcgcgt tgtccttttc cgctgcataa ccctgcttcg
     1561 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
     1621 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
     1681 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
     1741 cggtgctcaa cgggaatcct gctctgcgag gctggccgta ggccggccct accggcgcgg
     1801 cagcgttacc cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac acggctcatc
     1861 gggcatcggc aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg tcaaggctgg
     1921 caggtctggt tccatgcccg gaatgccggg ctggctgggc ggctcctcgc cggggccggt
     1981 cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc catgccccaa
     2041 cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca ccgacaggcg
     2101 caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg ccgacacggt
     2161 gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt ccttgactgc
     2221 gtattggacc gtccgcaaag aacgtccgat gagcttggaa agtgtcttct ggctgaccac
     2281 cacggcgttc tggtggccca tctgcgccac gaggtgatgc agcagcattg ccgccgtggg
     2341 tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt gcacccagtg
     2401 accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca tgcttatctc
     2461 catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca acttttcggc
     2521 agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt ttctttaacc
     2581 tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa
     2641 gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg tgcccaaaga
     2701 agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg
     2761 gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc
     2821 ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc
     2881 ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg
     2941 ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac cggcaagcga
     3001 agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa ttgatggggc
     3061 aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg ggtagccggt
     3121 gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc ttgtccagca
     3181 gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc catcgtccac
     3241 atatccacgg gctggcaagg gagcgcagcg accgcgcagg gc
//
