LOCUS       sequence-311198-        5715 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 5715)
  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 5715)
  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_PresTL-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    745..766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(781..786,787..804,805..811)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    819..835
                     /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     824..846
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     843..859
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     843..859
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        1083..1158
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1176..1193)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1176..1192)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1185..1207)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1759..2292
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2395..2983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2884..2903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagataagga ctactaattt atactgtaat ttattgtagc aagttgtggt tttactggtt
       61 ttgtttatat tagataagtt ttccatttat agctccttga ataccggatt ttttatctat
      121 aaagtatata tatcacaatt ttgatttaaa agtaaattct ttttttaaaa atacaaaaat
      181 tttttataaa ttatatagac catgatattg tattaatact aattgatttt aatttaaggt
      241 tgaactaaaa taaacatagt tttgtagaaa ataatttttc attattttgc tacttgatat
      301 tggatcgtaa aaatattagg gctttacaaa gttcttttaa aagagaattt agtaaagccc
      361 taagtctttt aataaaaaat tttttgtaaa aaagttggca aaaatagttt ttgctatata
      421 tttatatata agaaaaaatt caatactcaa ggagataaaa aatggaaaac ttaaacaata
      481 atccacaaaa aattaatcaa ataaataagc aaactaattt tattggtttt gaaaatttta
      541 tagaacttga taatcaagtt aatcaaacta aagaaaaaag taaattaagt aagataggta
      601 agaaattacc cggaataagc agtcaagtct agtgcggccg cgcccaatac gcaaaccgcc
      661 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      721 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      781 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      841 cacaggaaac agctatgaca tgattacgaa ttcgagctcc gaagtttatt attttatgat
      901 ttaaattaat aaaaagattt atcggtgtga atagaatatt tctttggttt attatgcatc
      961 ctagtacata ttatataatt taatttaata tataagcata tttttttaaa atcagaaatt
     1021 ttttacttag ctaaagattt taaacttggt ataattgaat tgaaaattta aatttctgac
     1081 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     1141 gtggcaccga gtcggtgctt tttttgaagc ttggcactgg ccgtcgtttt acaacgtcgt
     1201 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     1261 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1321 aatggcgaat ggcgcggccg ccctaggtta atacccgagc ttcaaggaag atttcctatt
     1381 aaggttgaac ttaagagctt aagcatagac gatttgaaaa aaattttaaa acaaacaaaa
     1441 aattctttaa taaaacaata tgttgcgatg tttaaggttt atgatttaga tttaaagttt
     1501 agtgaggaag ctatagatag aattgcagag cttactttta atatgaatct tgagagtgaa
     1561 aatcttggtg ccagaagact tcacggtgtt atggaaatag tgcttgcaga tctttttttt
     1621 gaagtgcctg gcagtaagtt gaaaaaattt gaaataaact tggactatgt taataaaaaa
     1681 atacaaatta acgaacaaaa agatttgaat tattatataa tttagttaaa agcaatttta
     1741 aatgagggag gtttccatat gttacgcagc agcaacgatg ttacgcagca gggcagtcgc
     1801 cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag gctcggccct
     1861 gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt cggagacgta
     1921 gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct ccgtagtaag
     1981 acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct cgcggcttac
     2041 gttctgccca agtttgagca gccgcgtagt gagatctata tctatgatct cgcagtctcc
     2101 ggcgagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa gcatgaggcc
     2161 aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga tcccgcagtg
     2221 gctctctata caaagttggg catacgggaa gaagtgatgc actttgatat cgacccaagt
     2281 accgccacct aacaattcgt tcaagccgag atcggcttac gcgtccaaaa tcccttaacg
     2341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcactaga
     3061 tccccggcaa gttcatcctt agggttttca tatctcttaa aaagtcaatt tctgcttgat
     3121 taatttcttg tggattattg tttttgcggt ttttcatttt ttatctcctt gagtattgaa
     3181 ttttttctta tatataaaat atagcaaaaa ctatttttgc caactttttt acaaaaaatt
     3241 ttttattaaa acacttaggg ctttgctaaa ttctctttta aaagaactta gtaaagccct
     3301 atggatttaa gaactgatta tacttacgta gtaaaaatac tacagatatt aatagtatag
     3361 accatatcaa aatcataatc aatttatatt tatctaaaat tgtggtccat acaaaataca
     3421 aaaaaattag gtttttctaa attctatttt aaaagaactt agtaacacct aatctctaaa
     3481 ttacttacaa aattctaaaa ttagattttt agttcttcat atttctttaa aagtttttta
     3541 agaaaatctt tttgattttc aaaaatttca tgcatcataa aactagtaaa tttggatttt
     3601 tttttgtaaa aatcataact ttcttgggtt ttaagttgaa atcttaatgg ttttattgga
     3661 ttttgcctag atttttttaa cgccggactt tctttatctt ttataaaatt ataagaattt
     3721 ttaataccat tttctattaa ataatcttct tttataagcc cctcttgcag tgcagttgct
     3781 aatcttaaat acttatacgc ttgagttttt gctacatcaa agtcttttat aaaatcaaga
     3841 aaagttttat aaccatcaaa tttataatat tgattatctt ttatttcttt taaaatttta
     3901 atattttcca ccctgtggaa tatttctttt ttgaaattgt catttaactt ttcctttagt
     3961 gtcttgtaat aagaaatttt actttcatta atcattaaat tttgttcaac ataagtatct
     4021 atcttttttg ttaaaacttt aacaacacgt tttttttcca taaaaatttt cctttatttt
     4081 tgcaacattt tccttaattg atataattgt tactatttta gattttttaa tatccactta
     4141 agaattaatc cctcactaga atatttttta ttatagaatt catatacatg tttttctaaa
     4201 tctataaatt tatcaacaat tgctttatta tagggtgtat ctatcttttc ttttcttaat
     4261 aatcgtctca tattctttaa ataacaacaa acactgccgg ttttaaacct aaattctatg
     4321 taatatattt ttgaaaaagt atatacattt tcaaccccgt ttatttcata ctttacaaaa
     4381 acattattta ttggttttct ataaccatag taaatgccta aaaatttgtc cccgtctttc
     4441 ataggatata gataaagttt ttgaatctct gatttatcta ataatttttc aaaaaaaatt
     4501 atacaccgct tttgtttgtt actaactact ctatgcaatg cctttaatat ttttgtgtga
     4561 tatatatttt tatcaccttt tttttctttc agcaggaaga tactttcttt tggaaattta
     4621 attttttgtt catttattct tgcagcctta agtccgattt tcatatttaa tccttatata
     4681 gcctttctat ttaattcttt taaactcaat aaactttttt ggtttttcat tagttctaat
     4741 aattcgcaat aatatgtgca aaacacttta ttgtattcta ttttcttttg gttattcaaa
     4801 tattctttta taattggctt tagaagttca atatttactt ctttttttag ttgttcaata
     4861 aggatattga aaatgtttgt ttttaaatct tgataatttt gttgagaatt ttcttttttt
     4921 ctttcaattg acttctctaa tttacgcttt atattgttta aatcgctata tttatgactt
     4981 tcaataataa aatgaggctt aaatttgtaa ttttcgtata ttttttggaa acttgtttct
     5041 aattgtttag aattatattc atttttttca aattcttttt gagtgttgca tagaatttcc
     5101 tttaattttt cttgtttttc tttcatacaa gatttagtaa aatgtatatt tttatttttt
     5161 agtaatttaa tttctattat ttttatgatt ttaagtattt tgattatatt atctttatca
     5221 atatttaaat tgaataaaat tgtaagaaaa attgaaagaa tttttttaca taaaaagtta
     5281 catttattga agtaatttct tacttgatac ttttctattt cattaatctt tctttcttct
     5341 tttatattat ttttattatt taaacactcc tttaaatcta cactcccatt tttagtaaat
     5401 ctatctttaa ggccccttga aactctattt tgaaatctta aatctttttt ttcttgaaag
     5461 tatttataga ttttttggta acattctttt tttgaaaaat taagttgata gtatatttcg
     5521 gtccccaaat ttacccctaa gtgtttgtga taattagttg tgactttaaa ttctttttct
     5581 aatttgtaaa ggtaattttg cagtgttttt agtttaacgg gtgtttgact atttctttta
     5641 aggttttcat taaagtaata gagtatgttt ttttgtgtgt atttcttata tgttttattt
     5701 atgtacgcta gtgtt
//
