LOCUS       sequence-306452-        6542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ftsI gRNA expression in B. burgdorferi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6542)
  AUTHORS   Takacs CN, Scott M, Chang Y, Kloos ZA, Irnov I, Rosa PA, Liu J, 
            Jacobs-Wagner C
  TITLE     A CRISPR interference platform for selective downregulation of gene 
            expression in Borrelia burgdorferi.
  JOURNAL   Appl Environ Microbiol. 2020 Nov 30. pii: AEM.02519-20. doi: 
            10.1128/AEM.02519-20.
  PUBMED    33257311
REFERENCE   2  (bases 1 to 6542)
  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: "Appl 
            Environ Microbiol. 2020 Nov 30. pii: AEM.02519-20. doi: 
            10.1128/AEM.02519-20."
COMMENT     Sequence Label: pBSV2_Psyn-sgRNAftsI
FEATURES             Location/Qualifiers
     source          1..6542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..38
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(53..58,59..76,77..83)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    91..107
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     96..118
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     115..131
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     115..131
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        168..202
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        224..299
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(342..359)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(342..358)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(351..373)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             915..1724
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(982..1001)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     regulatory      1975..1984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1981..2355
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      2493..3081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2982..3001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
       61 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      121 acagctatga ccatgattac gaattcgagc tcgaattcta aagatctttg acagctagct
      181 cagtcctagg tataatacta gtgagtgtta ttaagcattc tttgttttag agctagaaat
      241 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      301 ttttttgaag cttgggcccg aacaaaaact caaagcttgg cactggccgt cgttttacaa
      361 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
      421 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      481 agcctgaatg gcgaatggcg cggccgctac ccgagcttca aggaagattt cctattaagg
      541 ttgaacttaa gagcttaagc atagacgatt tgaaaaaaat tttaaaacaa acaaaaaatt
      601 ctttaataaa acaatatgtt gcgatgttta aggtttatga tttagattta aagtttagtg
      661 aggaagctat agatagaatt gcagagctta cttttaatat gaatcttgag agtgaaaatc
      721 ttggtgccag aagacttcac ggtgttatgg aaatagtgct tgcagatctt ttttttgaag
      781 tgcctggcag taagttgaaa aaatttgaaa taaacttgga ctatgttaat aaaaaaatac
      841 aaattaacga acaaaaagat ttgaattatt atataattta gttaaaagca attttaaatg
      901 agggaggttt ccatatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
      961 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1021 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1081 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1141 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1201 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1261 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1321 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     1381 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1441 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1501 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1561 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1621 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1681 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     1741 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcttac aatttcctga
     1801 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt
     1861 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1921 ccgctcatga gacaataacc ctgataaatg cttcaataat agcacgtgag gagggccacc
     1981 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     2041 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     2101 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     2161 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     2221 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     2281 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     2341 gaggagcagg actgacacgt gctaaaactt catttttaat ttaaaaggat ctaggtgaag
     2401 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2461 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2521 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2581 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2641 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2701 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2761 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2821 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2881 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2941 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3001 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3061 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3121 tgctggcctt ttgctcacat gttctttcct gcactagatc cccggcaagt tcatccttag
     3181 ggttttcata tctcttaaaa agtcaatttc tgcttgatta atttcttgtg gattattgtt
     3241 tttgcggttt ttcatttttt atctccttga gtattgaatt ttttcttata tataaatata
     3301 tagcaaaaac tatttttgcc aactttttta caaaaaattt tttattaaaa cacttagggc
     3361 tttgctaaat tctcttttaa aagaacttag taaagcccta tggatttaag aactgattat
     3421 acttacgtag taaaaatact acagatatta atagtataga ccatatcaaa atcataatca
     3481 atttatattt atctaaaatt gtagtccata caaaatacaa aaaaattagg tttttctaaa
     3541 ttctatttta aaagaactta gtaacaccta atctctaaat tacttacaaa attctaaaat
     3601 tagattttta gttcttcata tttctttaaa agttttttaa gaaaatcttt ttgattttca
     3661 aaaatttcat gcatcataaa actagtaaat ttggattttt ttttgtaaaa atcataactt
     3721 tcttgggttt taagttgaaa tcttaatggt tttattggat tttgcctaga tttttttaac
     3781 gccggacttt ctttatcttt tataaaatta taagaatttt taataccatt ttctattaaa
     3841 taatcttctt ttataagccc ctcttgcagt gcagttgcta atcttaaata cttatacgct
     3901 tgagtttttg ctacatcaaa gtcttttata aaatcaagaa aagttttata accatcaaat
     3961 ttataatatt gattatcttt tatttctttt aaaattttaa tattttccac cctgtggaat
     4021 atttcttttt tgaaattgtc atttaacttt tcctttagtg tcttgtaata agaaatttta
     4081 ctttcattaa tcattaaatt ttgttcaaca taagtatcta tcttttttgt taaaacttta
     4141 acaacacgtt ttttttccat aaaaattttc ctttattttt gcaacatttt ccttaattga
     4201 tataattgtt actattttag attttttaat atccacttaa gaattaatcc ctcactagaa
     4261 tattttttat tatagaattc atatacatgt ttttctaaat ctataaattt atcaacaatt
     4321 gctttattat agggtgtatc tatcttttct tttcttaata atcgtctcat attctttaaa
     4381 taacaacaaa cactgccggt tttaaaccta aattctatgt aatatatttt tgaaaaagta
     4441 tatacatttt caaccccgtt tatttcatac tttacaaaaa cattatttat tggttttcta
     4501 taaccatagt aaatgcctaa aaatttgtcc ccgtctttca taggatatag ataaagtttt
     4561 tgaatctctg atttatctaa taatttttca aaaaaaatta tacaccgctt ttgtttgtta
     4621 ctaactactc tatgcaatgc ctttaatatt tttgtgtgat atatattttt atcacctttt
     4681 ttttctttca gcaggaagat actttctttt ggaaatttaa ttttttgttc atttattctt
     4741 gcagccttaa gtccgatttt catatttaat ccttatatag cctttctatt taattctttt
     4801 aaactcaata aacttttttg gtttttcatt agttctaata attcgcaata atatgtgcaa
     4861 aacactttat tgtattctat tttcttttgg ttattcaaat attcttttat aattggcttt
     4921 agaagttcaa tatttacttc tttttttagt tgttcaataa ggatattgaa aatgtttgtt
     4981 tttaaatctt gataattttg ttgagaattt tctttttttc tttcaattga cttctctaat
     5041 ttacgcttta tattgtttaa atcgctatat ttatgacttt caataataaa atgaggctta
     5101 aatttgtaat tttcgtatat tttttggaaa cttgtttcta attgtttaga attatattca
     5161 tttttttcaa attctttttg agtgttgcat agaatttcct ttaatttttc ttgtttttct
     5221 ttcatacaag atttagtaaa atgtatattt ttatttttta gtaatttaat ttctattatt
     5281 tttatgattt taagtatttt gattatatta tctttatcaa tatttaaatt gaataaaatt
     5341 gtaagaaaaa ttgaaagaat ttttttacat aaaaagttac atttattgaa gtaatttctt
     5401 acttgatact tttctatttc attaatcttt ctttcttctt ttatattatt tttattattt
     5461 aaacactcct ttaaatctac actcccattt ttagtaaatc tatctttaag gccccttgaa
     5521 actctatttt gaaatcttaa atcttttttt tcttgaaagt atttatagat tttttggtaa
     5581 cattcttttt ttgaaaaatt aagttgatag tatatttcgg tccccaaatt tacccctaag
     5641 tgtttgtgat aattagttgt gactttaaat tctttttcta atttgtaaag gtaattttgc
     5701 agtgttttta gtttaacggg tgtttgacta tttcttttaa ggttttcatt aaagtaatag
     5761 agtatgtttt tttgtgtgta tttcttatat gttttattta tgtacgctag tgttgagata
     5821 aggactacta atttatactg taatttattg tagcaagttg tggttttact ggttttgttt
     5881 atattagata agttttccat ttatagctcc ttgaataccg gattttttat ctataaagta
     5941 tatatatcac aattttgatt taaaagtaaa ttcttttttt aaaaatacaa aaatttttta
     6001 taaattatat agaccatgat attgtattaa tactaattga ttttaattta aggttgaact
     6061 aaaataaaca tagttttgta gaaaataatt tttcattatt ttgctacttg atattggatc
     6121 gtaaaaatat tagggcttta caaagttctt ttaaaagaga atttagtaaa gccctaagtc
     6181 ttttaataaa aaattttttg taaaaaagtt ggcaaaaata gtttttgcta tatatttata
     6241 tataagaaaa aattcaatac tcaaggagat aaaaaatgga aaacttaaac aataatccac
     6301 aaaaaattaa tcaaataaat aagcaaacta attttattgg ttttgaaaat tttatagaac
     6361 ttgataatca agttaatcaa actaaagaaa aaagtaaatt aagtaagata ggtaagaaat
     6421 tacccggaat aagcagtcaa gtctagtgcg gccgcgccca atacgcaaac cgcctctccc
     6481 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6541 ca
//
