LOCUS       sequence-269536-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for constitutive expression of a single guide RNA from Streptococcus
            pasteurianus 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 5510)
  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 5510)
  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(Spa)_ctrA
FEATURES             Location/Qualifiers
     source          1..5510
                     /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      467..510
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(653..669)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(653..669)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(701..720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(702..720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(730..747)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(730..746)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(739..761)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1455..2117)
                     /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      2118..2889
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             4369..5163
                     /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(4423..4442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5033..5052
                     /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 gttaatttaa gactggttaa gtttttgtac tcgaaagagc ctacaaagat aaggctttat
      421 gccgaattca agcaccccat gttttgacat gaggtgcttt tttttgataa aacgaaaggc
      481 ccagtctttc gactgagcct ttcgttttat aaacgggtct tgaggggttt tttaagcttc
      541 tgcagctgga tccatggata tctagatttc cgcaaaataa aaaaggggac ctctagggtc
      601 cccaattaat tagctaaagc cgctagccca tgggaattcc tgcagcccgg gggatccact
      661 agttctagag cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta
      721 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      781 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      841 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt
      901 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
      961 gccgtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
     1021 ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc
     1081 gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
     1141 tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
     1201 gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcatt
     1261 tattctgcct cccagagcct gataaaaacg gtgaatccgt tagcgaggtg ccgccggctt
     1321 ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa
     1381 ggtatagggc ggcgaggcgg ctacagccga tagtctggaa cagcgcactt acgggttgct
     1441 gcgcaaccca agtgctaccg gcgcggcagc gtgacccgtg tcggcggctc caacggctcg
     1501 ccatcgtcca gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc
     1561 cattcctccg tttcggtcaa ggctggcagg tctggttcca tgcccggaat gccgggctgg
     1621 ctgggcggct cctcgccggg gccggtcggt agttgctgct cgcccggata cagggtcggg
     1681 atgcggcgca ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc
     1741 acggcggcac tgaacaccga caggcgcaac tggtcgcggg gctggcccca cgccacgcgg
     1801 tcattgacca cgtaggccga cacggtgccg gggccgttga gcttcacgac ggagatccag
     1861 cgctcggcca ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc
     1921 ttggaaagtg ttttctggct gaccaccacg gcgttctggt ggcccatctg cgccacgagg
     1981 tgatgcagca gcattgccgc cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc
     2041 gctttgcgtt ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat gccgatttct
     2101 ctggactgcg tggccatgct tatctccatg cggtaggggt gccgcacggt tgcggcacca
     2161 tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg
     2221 cagtcaatta cagattttct ttaacctacg caatgagcta ttgcgggggg tgccgcaatg
     2281 agctgttgcg tacccccctt ttttaagttg ttgattttta agtctttcgc atttcgccct
     2341 atatctagtt ctttggtgcc caaagaaggg cacccctgcg gggttccccc acgccttcgg
     2401 cgcggctccc cctccggcaa aaagtggccc ctccggggct tgttgatcga ctgcgcggcc
     2461 ttcggccttg cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc
     2521 tcggggggca ggcgggcggg cttcgccctt cgactgcccc cactcgcata ggcttgggtc
     2581 gttccaggcg cgtcaaggcc aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt
     2641 ccacttggtg tccaaccggc aagcgaagcg cgcaggccgc aggccggagg cttttcccca
     2701 gagaaaatta aaaaaattga tggggcaagg ccgcaggccg cgcagttgga gccggtgggt
     2761 atgtggtcga aggctgggta gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg
     2821 cagcctgtcc atcagcttgt ccagcagggt tgtccacggg ccgagcgaag cgagccagcc
     2881 ggtggccgct cgcggccatc gtccacatat ccacgggctg gcaagggagc gcagcgaccg
     2941 cgcagggcga agcccggaga gcaagcccgt agggcgccgc agccgccgta ggcggtcacg
     3001 actttgcgaa gcaaagtcta gtgagtatac tcaagcattg agtggcccgc cggaggcacc
     3061 gccttgcgct gcccccgtcg agccggttgg acaccaaaag ggaggggcag gcatggcggc
     3121 atacgcgatc atgcgatgca agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa
     3181 gcacgcctac cgcgagcgcg agacgcccaa cgctgacgcc agcaggacgc cagagaacga
     3241 gcactgggcg gccagcagca ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga
     3301 gaagcggcgc aaggacgctg tgttggcggt cgagtacgtc atgacggcca gcccggaatg
     3361 gtggaagtcg gccagccaag aacagcaggc ggcgttcttc gagaaggcgc acaagtggct
     3421 ggcggacaag tacggggcgg atcgcatcgt gacggccagc atccaccgtg acgaaaccag
     3481 cccgcacatg accgcgttcg tggtgccgct gacgcaggac ggcaggctgt cggccaagga
     3541 gttcatcggc aacaaagcgc agatgacccg cgaccagacc acgtttgcgg ccgctgtggc
     3601 cgatctaggg ctgcaacggg gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc
     3661 gttctacgag gccctggagc ggccaccagt gggccacgtc accatcagcc cgcaagcggt
     3721 cgagccacgc gcctatgcac cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt
     3781 tgagacgccg gaagccgtgg ccgaccggct gacaaaagcg gttcggcagg ggtatgagcc
     3841 tgccctacag gccgccgcag gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga
     3901 gacggcccga gaccttcggg agcgcctgaa gcccgttctg gacgccctgg ggccgttgaa
     3961 tcgggatatg caggccaagg ccgccgcgat catcaaggcc gtgggcgaaa agctgctgac
     4021 ggaacagcgg gaagtccagc gccagaaaca ggcccagcgc cagcaggaac gcgggcgcgc
     4081 acatttcccc gaaaagtgcc acctgggatg aatgtcagct actgggctat ctggacaagg
     4141 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     4201 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
     4261 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     4321 cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4381 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4441 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4501 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4561 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4621 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4681 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4741 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4801 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4861 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4921 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4981 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5041 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5101 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5161 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5221 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5281 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc cacccccatg
     5341 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     5401 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtttttatat
     5461 aagctttcag tcggcgcgaa tgctccacgc cgcgcccagc ggcgcggatc
//
