LOCUS       sequence-269541-        5516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for constitutive expression of a single guide RNA from Streptococcus
            thermophilus #3 with a seed region that targets ctrA gene's promoter
            from Caulobacter crescentus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5516)
  AUTHORS   Guzzo M, Castro LK, Reisch CR, Guo MS, Laub MT
  TITLE     A CRISPR Interference System for Efficient and Rapid Gene Knockdown 
            in Caulobacter crescentus.
  JOURNAL   mBio. 2020 Jan 14;11(1):e02415-19. doi: 10.1128/mBio.02415-19.
  PUBMED    31937638
REFERENCE   2  (bases 1 to 5516)
  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"; doi: "10.1128/mBio"; 
            journalName: "mBio"; date: "2020-01-14- 14"; volume: "11"; issue: 
            "1"; pages: "e02415-19"
COMMENT     Sequence Label: pBXMCS-2 Pconstitutive-sgRNA(Sth3)_ctrA
FEATURES             Location/Qualifiers
     source          1..5516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..47
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      473..516
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(659..675)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(659..675)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(707..726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(708..726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(736..753)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(736..752)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(745..767)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1461..2123)
                     /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      2124..2895
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             4375..5169
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4429..4448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5039..5058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
       61 ctgcagccag ccgtggtcgg gcagcaggta gaaggcgccc tcgtcctgat cctcgcccga
      121 aacctccagc ccccggtcga tggcttcgac gacatagccg gccgcgcggc aggtgtcggt
      181 gagcgcggcc agcagggcgg cttcctggtc aggggtcagg tcggtcatgg gcaagaggtc
      241 caggtcgtgg tttgtcggcg gcttctagca tggaccgccc gcgcccgtga ggccgaggat
      301 ttcgcgctgg tcagacaacc tacttgccgt ccccacatgt tagcgctacc aagtgccgac
      361 acgtctttgg cgggagtccg cgttttagag ctgtgaaaac agcgagttaa aataaggctt
      421 agtccgtact caacttgaaa aggtggcacc gattcggtgt tttttttttt tgataaaacg
      481 aaaggcccag tctttcgact gagcctttcg ttttataaac gggtcttgag gggtttttta
      541 agcttctgca gctggatcca tggatatcta gatttccgca aaataaaaaa ggggacctct
      601 agggtcccca attaattagc taaagccgct agcccatggg aattcctgca gcccggggga
      661 tccactagtt ctagagcggc cgccaccgcg gtggagctcc aattcgccct atagtgagtc
      721 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      781 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      841 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
      901 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
      961 caataggccg tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
     1021 gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga tgaatggcag
     1081 aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta aatagccgct
     1141 tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgac cgtgtgcttc
     1201 tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat tgacgatatg
     1261 atcatttatt ctgcctccca gagcctgata aaaacggtga atccgttagc gaggtgccgc
     1321 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     1381 agacaaggta tagggcggcg aggcggctac agccgatagt ctggaacagc gcacttacgg
     1441 gttgctgcgc aacccaagtg ctaccggcgc ggcagcgtga cccgtgtcgg cggctccaac
     1501 ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc
     1561 cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg
     1621 ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc cggatacagg
     1681 gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca
     1741 accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg gccccacgcc
     1801 acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt cacgacggag
     1861 atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa agaacgtccg
     1921 atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc catctgcgcc
     1981 acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc ggcccacgcc
     2041 tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc ttgaatgccg
     2101 atttctctgg actgcgtggc catgcttatc tccatgcggt aggggtgccg cacggttgcg
     2161 gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg cgttgcgtaa
     2221 aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc ggggggtgcc
     2281 gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc tttcgcattt
     2341 cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc
     2401 cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc
     2461 gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca ctcgccgccg
     2521 tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac tgcccccact cgcataggct
     2581 tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc
     2641 cgccttccac ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc cggatgcttt
     2701 tccccagaga aaattaaaaa aattgatggg gcaaggccgc aggccgcgca gttggagccg
     2761 gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca agctcgtggg
     2821 caggcgcagc ctgtccatca gcttgtccag cagggttgtc cacgtgccga gcgaagcgag
     2881 ccagccggtg gccgctcgcg gccatcgtcc acatatccac gggctggcaa gggagcgcag
     2941 cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg cgccgcagcc gccgtaggcg
     3001 gtcatgactt tgcgaagcaa agtctagtga gtatactcaa gcattgagtg gcccgccgga
     3061 ggcaccgcct tgcgctgccc ccgtcgagcc ggttggacac caaaagggag gggcaggcat
     3121 ggcggcatac gcgatcatgc gatgcaagaa gctggcgaaa atgtgcaacg tggcggccag
     3181 tctcaagcac gcctaccgcg agcgcgagac gcccaacgct gacgccagca ggacgccaga
     3241 gaacgagcac tgggcggcca gcagcaccga tgaagcgatg ggccgactgc gcgagttgct
     3301 gccagagaag cggcgcaagg acgctgtgtt ggcggtcgag tacgtcatga cggccagccc
     3361 ggaatggtgg aagtcggcca gccaagaaca gcaggcggcg ttcttcgaga aggcgcacaa
     3421 gtggctggcg gacaagtacg gggcggatcg catcgtgacg gccagcatcc accgtgacga
     3481 aacaagcccg cacatgaccg cgttcgtggt gccgctgacg caggacggca ggctgtcggc
     3541 caaggagttc atcggcaaca aagcgcagat gacccgcgac cagaccacgt ttgcggccgc
     3601 tgtggccgat ctagggctgc aacggggcat cgagggcagc aaggcacgtc acacgcgcat
     3661 tcaggcgttc tacgaggccc tggagcggcc accagtgggc cacgtcacca tcagcccgca
     3721 agcggtcgag ccacgcgcct atgcaccgca gggattggcc gaaaagctgg gaatctcaaa
     3781 gcgcgttgag acgccggaag ccgtggccga ccggctgaca aaagcggttc ggcaggggta
     3841 tgagcctgcc ctacaggccg ccgcaggagc gcgtgagatg cgcaagaagg ccgatcaagc
     3901 ccaagagacg gcccgagacc ttcgggagcg cctgaagccc gttctggacg ccctggggcc
     3961 gttgaatcgg gatatgcagg ccaaggccgc cgcgatcatc aaggccgtgg gcgaaaagct
     4021 gctgacggaa cagcgggaag tccagcgcca gaaacaggcc cagcgccagc aggaacgcgg
     4081 gcgcgcacat ttccccgaaa agtgccacct gggatgaatg tcagctactg ggctatctgg
     4141 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     4201 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     4261 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     4321 tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4381 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4441 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4501 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4561 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4621 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4681 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4741 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4801 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4861 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4921 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4981 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5041 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5101 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5161 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5221 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5281 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5341 cccatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     5401 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagttt
     5461 ttatataagc tttcagtcgg cgcgaatgct ccacgccgcg cccagcggcg cggatc
//
