LOCUS       sequence-311188-        5831 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 5831)
  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 5831)
  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_PflaBS-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    753..774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(789..794,795..812,813..819)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    827..843
                     /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     832..854
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     851..867
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     851..867
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        1207..1282
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1300..1317)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1300..1316)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1309..1331)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1883..2416
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2519..3107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3008..3027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagtgttga gataaggact actaatttat actgtaattt attgtagcaa gttgtggttt
       61 tactggtttt gtttatatta gataagtttt ccatttatag ctccttgaat accggatttt
      121 ttatctataa agtatatata tcacaatttt gatttaaaag taaattcttt ttttaaaaat
      181 acaaaaattt tttataaatt atatagacca tgatattgta ttaatactaa ttgattttaa
      241 tttaaggttg aactaaaata aacatagttt tgtagaaaat aatttttcat tattttgcta
      301 cttgatattg gatcgtaaaa atattagggc tttacaaagt tcttttaaaa gagaatttag
      361 taaagcccta agtcttttaa taaaaaattt tttgtaaaaa agttggcaaa aatagttttt
      421 gctatatatt tatatataag aaaaaattca atactcaagg agataaaaaa tggaaaactt
      481 aaacaataat ccacaaaaaa ttaatcaaat aaataagcaa actaatttta ttggttttga
      541 aaattttata gaacttgata atcaagttaa tcaaactaaa gaaaaaagta aattaagtaa
      601 gataggtaag aaattacccg gaataagcag tcaagtctag tgcggccgcg cccaatacgc
      661 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
      721 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      781 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      841 caatttcaca caggaaacag ctatgacatg attacgaatt cgagctctgt ctgtcgcctc
      901 ttgtggcttc cggaggagtt atttataata aaataaagaa ttacgacttg aaaattttta
      961 atcaaaaaaa taaagttgaa gattataaaa agaagtacga agatagagag agaaaagtgg
     1021 aaggtgaact taataccttg gattttaccg ttaagcgcat gaaagatcaa gaaaatacat
     1081 taaaggcttt tgattttaat caaagaaata aataataata attattttta atgctattgc
     1141 tatttgcgtt tctttttttt taatttttgt gctattcttt ttaacgaaaa tttaaatttc
     1201 tgacttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     1261 aaaagtggca ccgagtcggt gctttttttg aagcttggca ctggccgtcg ttttacaacg
     1321 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1381 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1441 cctgaatggc gaatggcgcg gccgccctag gttaataccc gagcttcaag gaagatttcc
     1501 tattaaggtt gaacttaaga gcttaagcat agacgatttg aaaaaaattt taaaacaaac
     1561 aaaaaattct ttaataaaac aatatgttgc gatgtttaag gtttatgatt tagatttaaa
     1621 gtttagtgag gaagctatag atagaattgc agagcttact tttaatatga atcttgagag
     1681 tgaaaatctt ggtgccagaa gacttcacgg tgttatggaa atagtgcttg cagatctttt
     1741 ttttgaagtg cctggcagta agttgaaaaa atttgaaata aacttggact atgttaataa
     1801 aaaaatacaa attaacgaac aaaaagattt gaattattat ataatttagt taaaagcaat
     1861 tttaaatgag ggaggtttcc atatgttacg cagcagcaac gatgttacgc agcagggcag
     1921 tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat gtaggctcgg
     1981 ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga
     2041 cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact tgctccgtag
     2101 taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc
     2161 ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg atctcgcagt
     2221 ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc tcaagcatga
     2281 ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg acgatcccgc
     2341 agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg atatcgaccc
     2401 aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttacgcgtcc aaaatccctt
     2461 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2521 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2581 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2641 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2701 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2761 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2821 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2881 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2941 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3001 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3061 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3121 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcac
     3181 tagatccccg gcaagttcat ccttagggtt ttcatatctc ttaaaaagtc aatttctgct
     3241 tgattaattt cttgtggatt attgtttttg cggtttttca ttttttatct ccttgagtat
     3301 tgaatttttt cttatatata aaatatagca aaaactattt ttgccaactt ttttacaaaa
     3361 aattttttat taaaacactt agggctttgc taaattctct tttaaaagaa cttagtaaag
     3421 ccctatggat ttaagaactg attatactta cgtagtaaaa atactacaga tattaatagt
     3481 atagaccata tcaaaatcat aatcaattta tatttatcta aaattgtggt ccatacaaaa
     3541 tacaaaaaaa ttaggttttt ctaaattcta ttttaaaaga acttagtaac acctaatctc
     3601 taaattactt acaaaattct aaaattagat ttttagttct tcatatttct ttaaaagttt
     3661 tttaagaaaa tctttttgat tttcaaaaat ttcatgcatc ataaaactag taaatttgga
     3721 tttttttttg taaaaatcat aactttcttg ggttttaagt tgaaatctta atggttttat
     3781 tggattttgc ctagattttt ttaacgccgg actttcttta tcttttataa aattataaga
     3841 atttttaata ccattttcta ttaaataatc ttcttttata agcccctctt gcagtgcagt
     3901 tgctaatctt aaatacttat acgcttgagt ttttgctaca tcaaagtctt ttataaaatc
     3961 aagaaaagtt ttataaccat caaatttata atattgatta tcttttattt cttttaaaat
     4021 tttaatattt tccaccctgt ggaatatttc ttttttgaaa ttgtcattta acttttcctt
     4081 tagtgtcttg taataagaaa ttttactttc attaatcatt aaattttgtt caacataagt
     4141 atctatcttt tttgttaaaa ctttaacaac acgttttttt tccataaaaa ttttccttta
     4201 tttttgcaac attttcctta attgatataa ttgttactat tttagatttt ttaatatcca
     4261 cttaagaatt aatccctcac tagaatattt tttattatag aattcatata catgtttttc
     4321 taaatctata aatttatcaa caattgcttt attatagggt gtatctatct tttcttttct
     4381 taataatcgt ctcatattct ttaaataaca acaaacactg ccggttttaa acctaaattc
     4441 tatgtaatat atttttgaaa aagtatatac attttcaacc ccgtttattt catactttac
     4501 aaaaacatta tttattggtt ttctataacc atagtaaatg cctaaaaatt tgtccccgtc
     4561 tttcatagga tatagataaa gtttttgaat ctctgattta tctaataatt tttcaaaaaa
     4621 aattatacac cgcttttgtt tgttactaac tactctatgc aatgccttta atatttttgt
     4681 gtgatatata tttttatcac cttttttttc tttcagcagg aagatacttt cttttggaaa
     4741 tttaattttt tgttcattta ttcttgcagc cttaagtccg attttcatat ttaatcctta
     4801 tatagccttt ctatttaatt cttttaaact caataaactt ttttggtttt tcattagttc
     4861 taataattcg caataatatg tgcaaaacac tttattgtat tctattttct tttggttatt
     4921 caaatattct tttataattg gctttagaag ttcaatattt acttcttttt ttagttgttc
     4981 aataaggata ttgaaaatgt ttgtttttaa atcttgataa ttttgttgag aattttcttt
     5041 ttttctttca attgacttct ctaatttacg ctttatattg tttaaatcgc tatatttatg
     5101 actttcaata ataaaatgag gcttaaattt gtaattttcg tatatttttt ggaaacttgt
     5161 ttctaattgt ttagaattat attcattttt ttcaaattct ttttgagtgt tgcatagaat
     5221 ttcctttaat ttttcttgtt tttctttcat acaagattta gtaaaatgta tatttttatt
     5281 ttttagtaat ttaatttcta ttatttttat gattttaagt attttgatta tattatcttt
     5341 atcaatattt aaattgaata aaattgtaag aaaaattgaa agaatttttt tacataaaaa
     5401 gttacattta ttgaagtaat ttcttacttg atacttttct atttcattaa tctttctttc
     5461 ttcttttata ttatttttat tatttaaaca ctcctttaaa tctacactcc catttttagt
     5521 aaatctatct ttaaggcccc ttgaaactct attttgaaat cttaaatctt ttttttcttg
     5581 aaagtattta tagatttttt ggtaacattc tttttttgaa aaattaagtt gatagtatat
     5641 ttcggtcccc aaatttaccc ctaagtgttt gtgataatta gttgtgactt taaattcttt
     5701 ttctaatttg taaaggtaat tttgcagtgt ttttagttta acgggtgttt gactatttct
     5761 tttaaggttt tcattaaagt aatagagtat gtttttttgt gtgtatttct tatatgtttt
     5821 atttatgtac g
//
