LOCUS       sequence-306829-        5608 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 5608)
  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 5608)
  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_Psyn-sgRNAftsI
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    282..303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(318..323,324..341,342..348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    356..372
                     /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     361..383
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     380..396
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     380..396
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        432..466
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        488..563
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(606..623)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(606..622)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(615..637)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1189..1722
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1825..2413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2314..2333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaaaactta aacaataatc cacaaaaaat taatcaaata aataagcaaa ctaattttat
       61 tggttttgaa aattttatag aacttgataa tcaagttaat caaactaaag aaaaaagtaa
      121 attaagtaag ataggtaaga aattacccgg aataagcagt caagtctagt gcggccgcgc
      181 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
      241 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      301 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      361 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattc gagctcgaat
      421 tctaaagatc tttgacagct agctcagtcc taggtataat actagtgagt gttattaagc
      481 attctttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      541 aaaaagtggc accgagtcgg tgcttttttt gaagcttggg cccgaacaaa aactcaaagc
      601 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      661 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      721 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcggccg ccctaggtta
      781 atacccgagc ttcaaggaag atttcctatt aaggttgaac ttaagagctt aagcatagac
      841 gatttgaaaa aaattttaaa acaaacaaaa aattctttaa taaaacaata tgttgcgatg
      901 tttaaggttt atgatttaga tttaaagttt agtgaggaag ctatagatag aattgcagag
      961 cttactttta atatgaatct tgagagtgaa aatcttggtg ccagaagact tcacggtgtt
     1021 atggaaatag tgcttgcaga tctttttttt gaagtgcctg gcagtaagtt gaaaaaattt
     1081 gaaataaact tggactatgt taataaaaaa atacaaatta acgaacaaaa agatttgaat
     1141 tattatataa tttagttaaa agcaatttta aatgagggag gtttccatat gttacgcagc
     1201 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg
     1261 ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg ggctgctctt
     1321 gatcttttcg gtcgtgagtt cggagacgta gccacctact cccaacatca gccggactcc
     1381 gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa
     1441 gaagcggttg ttggcgctct cgcggcttac gttctgccca agtttgagca gccgcgtagt
     1501 gagatctata tctatgatct cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc
     1561 gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg
     1621 caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg catacgggaa
     1681 gaagtgatgc actttgatat cgacccaagt accgccacct aacaattcgt tcaagccgag
     1741 atcggcttac gcgtccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1801 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1861 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1921 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1981 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2041 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2101 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2161 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2221 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2281 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2341 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2401 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2461 ttttgctcac atgttctttc ctgcactaga tccccggcaa gttcatcctt agggttttca
     2521 tatctcttaa aaagtcaatt tctgcttgat taatttcttg tggattattg tttttgcggt
     2581 ttttcatttt ttatctcctt gagtattgaa ttttttctta tatataaaat atagcaaaaa
     2641 ctatttttgc caactttttt acaaaaaatt ttttattaaa acacttaggg ctttgctaaa
     2701 ttctctttta aaagaactta gtaaagccct atggatttaa gaactgatta tacttacgta
     2761 gtaaaaatac tacagatatt aatagtatag accatatcaa aatcataatc aatttatatt
     2821 tatctaaaat tgtggtccat acaaaataca aaaaaattag gtttttctaa attctatttt
     2881 aaaagaactt agtaacacct aatctctaaa ttacttacaa aattctaaaa ttagattttt
     2941 agttcttcat atttctttaa aagtttttta agaaaatctt tttgattttc aaaaatttca
     3001 tgcatcataa aactagtaaa tttggatttt tttttgtaaa aatcataact ttcttgggtt
     3061 ttaagttgaa atcttaatgg ttttattgga ttttgcctag atttttttaa cgccggactt
     3121 tctttatctt ttataaaatt ataagaattt ttaataccat tttctattaa ataatcttct
     3181 tttataagcc cctcttgcag tgcagttgct aatcttaaat acttatacgc ttgagttttt
     3241 gctacatcaa agtcttttat aaaatcaaga aaagttttat aaccatcaaa tttataatat
     3301 tgattatctt ttatttcttt taaaatttta atattttcca ccctgtggaa tatttctttt
     3361 ttgaaattgt catttaactt ttcctttagt gtcttgtaat aagaaatttt actttcatta
     3421 atcattaaat tttgttcaac ataagtatct atcttttttg ttaaaacttt aacaacacgt
     3481 tttttttcca taaaaatttt cctttatttt tgcaacattt tccttaattg atataattgt
     3541 tactatttta gattttttaa tatccactta agaattaatc cctcactaga atatttttta
     3601 ttatagaatt catatacatg tttttctaaa tctataaatt tatcaacaat tgctttatta
     3661 tagggtgtat ctatcttttc ttttcttaat aatcgtctca tattctttaa ataacaacaa
     3721 acactgccgg ttttaaacct aaattctatg taatatattt ttgaaaaagt atatacattt
     3781 tcaaccccgt ttatttcata ctttacaaaa acattattta ttggttttct ataaccatag
     3841 taaatgccta aaaatttgtc cccgtctttc ataggatata gataaagttt ttgaatctct
     3901 gatttatcta ataatttttc aaaaaaaatt atacaccgct tttgtttgtt actaactact
     3961 ctatgcaatg cctttaatat ttttgtgtga tatatatttt tatcaccttt tttttctttc
     4021 agcaggaaga tactttcttt tggaaattta attttttgtt catttattct tgcagcctta
     4081 agtccgattt tcatatttaa tccttatata gcctttctat ttaattcttt taaactcaat
     4141 aaactttttt ggtttttcat tagttctaat aattcgcaat aatatgtgca aaacacttta
     4201 ttgtattcta ttttcttttg gttattcaaa tattctttta taattggctt tagaagttca
     4261 atatttactt ctttttttag ttgttcaata aggatattga aaatgtttgt ttttaaatct
     4321 tgataatttt gttgagaatt ttcttttttt ctttcaattg acttctctaa tttacgcttt
     4381 atattgttta aatcgctata tttatgactt tcaataataa aatgaggctt aaatttgtaa
     4441 ttttcgtata ttttttggaa acttgtttct aattgtttag aattatattc atttttttca
     4501 aattcttttt gagtgttgca tagaatttcc tttaattttt cttgtttttc tttcatacaa
     4561 gatttagtaa aatgtatatt tttatttttt agtaatttaa tttctattat ttttatgatt
     4621 ttaagtattt tgattatatt atctttatca atatttaaat tgaataaaat tgtaagaaaa
     4681 attgaaagaa tttttttaca taaaaagtta catttattga agtaatttct tacttgatac
     4741 ttttctattt cattaatctt tctttcttct tttatattat ttttattatt taaacactcc
     4801 tttaaatcta cactcccatt tttagtaaat ctatctttaa ggccccttga aactctattt
     4861 tgaaatctta aatctttttt ttcttgaaag tatttataga ttttttggta acattctttt
     4921 tttgaaaaat taagttgata gtatatttcg gtccccaaat ttacccctaa gtgtttgtga
     4981 taattagttg tgactttaaa ttctttttct aatttgtaaa ggtaattttg cagtgttttt
     5041 agtttaacgg gtgtttgact atttctttta aggttttcat taaagtaata gagtatgttt
     5101 ttttgtgtgt atttcttata tgttttattt atgtacgcta gtgttgagat aaggactact
     5161 aatttatact gtaatttatt gtagcaagtt gtggttttac tggttttgtt tatattagat
     5221 aagttttcca tttatagctc cttgaatacc ggatttttta tctataaagt atatatatca
     5281 caattttgat ttaaaagtaa attctttttt taaaaataca aaaatttttt ataaattata
     5341 tagaccatga tattgtatta atactaattg attttaattt aaggttgaac taaaataaac
     5401 atagttttgt agaaaataat ttttcattat tttgctactt gatattggat cgtaaaaata
     5461 ttagggcttt acaaagttct tttaaaagag aatttagtaa agccctaagt cttttaataa
     5521 aaaatttttt gtaaaaaagt tggcaaaaat agtttttgct atatatttat atataagaaa
     5581 aaattcaata ctcaaggaga taaaaaat
//
