LOCUS       sequence-311493-        5714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flaB gRNA expression in B. burgdorferi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  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 5714)
  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: pBSV2G_P0526-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        223..298
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(316..333)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(316..332)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(325..347)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             899..1432
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1535..2123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2024..2043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5600..5621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5636..5641,5642..5659,5660..5666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5674..5690
                     /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     5679..5701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5698..5714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5698..5714
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgattacga attcgagctc cttgttttca atgataggtt ttttaggtag agacttgtaa
       61 gggtttgttc ctggaatata atttttagga attttcatat attttactcc tatttaacta
      121 gctaacctat aattctatta ttttttttgt ttttatgcaa tcaatattga tattttgatt
      181 ttttatgatt agaatcatca tgaaaattta aatttctgac ttgttttaga gctagaaata
      241 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      301 tttttgaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      361 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      421 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcggccg
      481 ccctaggtta atacccgagc ttcaaggaag atttcctatt aaggttgaac ttaagagctt
      541 aagcatagac gatttgaaaa aaattttaaa acaaacaaaa aattctttaa taaaacaata
      601 tgttgcgatg tttaaggttt atgatttaga tttaaagttt agtgaggaag ctatagatag
      661 aattgcagag cttactttta atatgaatct tgagagtgaa aatcttggtg ccagaagact
      721 tcacggtgtt atggaaatag tgcttgcaga tctttttttt gaagtgcctg gcagtaagtt
      781 gaaaaaattt gaaataaact tggactatgt taataaaaaa atacaaatta acgaacaaaa
      841 agatttgaat tattatataa tttagttaaa agcaatttta aatgagggag gtttccatat
      901 gttacgcagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg
      961 ctcaagtatg ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg
     1021 ggctgctctt gatcttttcg gtcgtgagtt cggagacgta gccacctact cccaacatca
     1081 gccggactcc gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc
     1141 cttcgaccaa gaagcggttg ttggcgctct cgcggcttac gttctgccca agtttgagca
     1201 gccgcgtagt gagatctata tctatgatct cgcagtctcc ggcgagcacc ggaggcaggg
     1261 cattgccacc gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt
     1321 gatctacgtg caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg
     1381 catacgggaa gaagtgatgc actttgatat cgacccaagt accgccacct aacaattcgt
     1441 tcaagccgag atcggcttac gcgtccaaaa tcccttaacg tgagttttcg ttccactgag
     1501 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1561 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1621 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1681 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1741 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1801 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1861 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1921 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1981 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2041 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2101 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2161 tttgctggcc ttttgctcac atgttctttc ctgcactaga tccccggcaa gttcatcctt
     2221 agggttttca tatctcttaa aaagtcaatt tctgcttgat taatttcttg tggattattg
     2281 tttttgcggt ttttcatttt ttatctcctt gagtattgaa ttttttctta tatataaaat
     2341 atagcaaaaa ctatttttgc caactttttt acaaaaaatt ttttattaaa acacttaggg
     2401 ctttgctaaa ttctctttta aaagaactta gtaaagccct atggatttaa gaactgatta
     2461 tacttacgta gtaaaaatac tacagatatt aatagtatag accatatcaa aatcataatc
     2521 aatttatatt tatctaaaat tgtggtccat acaaaataca aaaaaattag gtttttctaa
     2581 attctatttt aaaagaactt agtaacacct aatctctaaa ttacttacaa aattctaaaa
     2641 ttagattttt agttcttcat atttctttaa aagtttttta agaaaatctt tttgattttc
     2701 aaaaatttca tgcatcataa aactagtaaa tttggatttt tttttgtaaa aatcataact
     2761 ttcttgggtt ttaagttgaa atcttaatgg ttttattgga ttttgcctag atttttttaa
     2821 cgccggactt tctttatctt ttataaaatt ataagaattt ttaataccat tttctattaa
     2881 ataatcttct tttataagcc cctcttgcag tgcagttgct aatcttaaat acttatacgc
     2941 ttgagttttt gctacatcaa agtcttttat aaaatcaaga aaagttttat aaccatcaaa
     3001 tttataatat tgattatctt ttatttcttt taaaatttta atattttcca ccctgtggaa
     3061 tatttctttt ttgaaattgt catttaactt ttcctttagt gtcttgtaat aagaaatttt
     3121 actttcatta atcattaaat tttgttcaac ataagtatct atcttttttg ttaaaacttt
     3181 aacaacacgt tttttttcca taaaaatttt cctttatttt tgcaacattt tccttaattg
     3241 atataattgt tactatttta gattttttaa tatccactta agaattaatc cctcactaga
     3301 atatttttta ttatagaatt catatacatg tttttctaaa tctataaatt tatcaacaat
     3361 tgctttatta tagggtgtat ctatcttttc ttttcttaat aatcgtctca tattctttaa
     3421 ataacaacaa acactgccgg ttttaaacct aaattctatg taatatattt ttgaaaaagt
     3481 atatacattt tcaaccccgt ttatttcata ctttacaaaa acattattta ttggttttct
     3541 ataaccatag taaatgccta aaaatttgtc cccgtctttc ataggatata gataaagttt
     3601 ttgaatctct gatttatcta ataatttttc aaaaaaaatt atacaccgct tttgtttgtt
     3661 actaactact ctatgcaatg cctttaatat ttttgtgtga tatatatttt tatcaccttt
     3721 tttttctttc agcaggaaga tactttcttt tggaaattta attttttgtt catttattct
     3781 tgcagcctta agtccgattt tcatatttaa tccttatata gcctttctat ttaattcttt
     3841 taaactcaat aaactttttt ggtttttcat tagttctaat aattcgcaat aatatgtgca
     3901 aaacacttta ttgtattcta ttttcttttg gttattcaaa tattctttta taattggctt
     3961 tagaagttca atatttactt ctttttttag ttgttcaata aggatattga aaatgtttgt
     4021 ttttaaatct tgataatttt gttgagaatt ttcttttttt ctttcaattg acttctctaa
     4081 tttacgcttt atattgttta aatcgctata tttatgactt tcaataataa aatgaggctt
     4141 aaatttgtaa ttttcgtata ttttttggaa acttgtttct aattgtttag aattatattc
     4201 atttttttca aattcttttt gagtgttgca tagaatttcc tttaattttt cttgtttttc
     4261 tttcatacaa gatttagtaa aatgtatatt tttatttttt agtaatttaa tttctattat
     4321 ttttatgatt ttaagtattt tgattatatt atctttatca atatttaaat tgaataaaat
     4381 tgtaagaaaa attgaaagaa tttttttaca taaaaagtta catttattga agtaatttct
     4441 tacttgatac ttttctattt cattaatctt tctttcttct tttatattat ttttattatt
     4501 taaacactcc tttaaatcta cactcccatt tttagtaaat ctatctttaa ggccccttga
     4561 aactctattt tgaaatctta aatctttttt ttcttgaaag tatttataga ttttttggta
     4621 acattctttt tttgaaaaat taagttgata gtatatttcg gtccccaaat ttacccctaa
     4681 gtgtttgtga taattagttg tgactttaaa ttctttttct aatttgtaaa ggtaattttg
     4741 cagtgttttt agtttaacgg gtgtttgact atttctttta aggttttcat taaagtaata
     4801 gagtatgttt ttttgtgtgt atttcttata tgttttattt atgtacgcta gtgttgagat
     4861 aaggactact aatttatact gtaatttatt gtagcaagtt gtggttttac tggttttgtt
     4921 tatattagat aagttttcca tttatagctc cttgaatacc ggatttttta tctataaagt
     4981 atatatatca caattttgat ttaaaagtaa attctttttt taaaaataca aaaatttttt
     5041 ataaattata tagaccatga tattgtatta atactaattg attttaattt aaggttgaac
     5101 taaaataaac atagttttgt agaaaataat ttttcattat tttgctactt gatattggat
     5161 cgtaaaaata ttagggcttt acaaagttct tttaaaagag aatttagtaa agccctaagt
     5221 cttttaataa aaaatttttt gtaaaaaagt tggcaaaaat agtttttgct atatatttat
     5281 atataagaaa aaattcaata ctcaaggaga taaaaaatgg aaaacttaaa caataatcca
     5341 caaaaaatta atcaaataaa taagcaaact aattttattg gttttgaaaa ttttatagaa
     5401 cttgataatc aagttaatca aactaaagaa aaaagtaaat taagtaagat aggtaagaaa
     5461 ttacccggaa taagcagtca agtctagtgc ggccgcgccc aatacgcaaa ccgcctctcc
     5521 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5581 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5641 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5701 gaaacagcta tgac
//
