LOCUS       sequence-311094-        5717 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 5717)
  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 5717)
  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_P0031-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1165..1186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1201..1206,1207..1224,1225..1231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1239..1255
                     /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     1244..1266
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1263..1279
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1263..1279
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        1505..1580
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1598..1615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1598..1614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2181..2714
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2817..3405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3306..3325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttcttactt gatacttttc tatttcatta atctttcttt cttcttttat attattttta
       61 ttatttaaac actcctttaa atctacactc ccatttttag taaatctatc tttaaggccc
      121 cttgaaactc tattttgaaa tcttaaatct tttttttctt gaaagtattt atagattttt
      181 tggtaacatt ctttttttga aaaattaagt tgatagtata tttcggtccc caaatttacc
      241 cctaagtgtt tgtgataatt agttgtgact ttaaattctt tttctaattt gtaaaggtaa
      301 ttttgcagtg tttttagttt aacgggtgtt tgactatttc ttttaaggtt ttcattaaag
      361 taatagagta tgtttttttg tgtgtatttc ttatatgttt tatttatgta cgctagtgtt
      421 gagataagga ctactaattt atactgtaat ttattgtagc aagttgtggt tttactggtt
      481 ttgtttatat tagataagtt ttccatttat agctccttga ataccggatt ttttatctat
      541 aaagtatata tatcacaatt ttgatttaaa agtaaattct ttttttaaaa atacaaaaat
      601 tttttataaa ttatatagac catgatattg tattaatact aattgatttt aatttaaggt
      661 tgaactaaaa taaacatagt tttgtagaaa ataatttttc attattttgc tacttgatat
      721 tggatcgtaa aaatattagg gctttacaaa gttcttttaa aagagaattt agtaaagccc
      781 taagtctttt aataaaaaat tttttgtaaa aaagttggca aaaatagttt ttgctatata
      841 tttatatata agaaaaaatt caatactcaa ggagataaaa aatggaaaac ttaaacaata
      901 atccacaaaa aattaatcaa ataaataagc aaactaattt tattggtttt gaaaatttta
      961 tagaacttga taatcaagtt aatcaaacta aagaaaaaag taaattaagt aagataggta
     1021 agaaattacc cggaataagc agtcaagtct agtgcggccg cgcccaatac gcaaaccgcc
     1081 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1141 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     1201 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     1261 cacaggaaac agctatgaca tgattacgaa ttcgagctcg ttgatattaa acttaaaagc
     1321 aatattattg ttgtaagaaa tacgtaaacc ttaggctcta ttcctaggat tggctttaat
     1381 ttaacttcaa aaagctcaac atttgaattc tcgtcaacct ctttcatctt ttaaagtata
     1441 gaaaagtatt taaataagtg tcaatatttt gtattattta attgaaaatt taaatttctg
     1501 acttgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     1561 aagtggcacc gagtcggtgc tttttttgaa gcttggcact ggccgtcgtt ttacaacgtc
     1621 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1681 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1741 tgaatggcga atggcgcggc cgccctaggt taatacccga gcttcaagga agatttccta
     1801 ttaaggttga acttaagagc ttaagcatag acgatttgaa aaaaatttta aaacaaacaa
     1861 aaaattcttt aataaaacaa tatgttgcga tgtttaaggt ttatgattta gatttaaagt
     1921 ttagtgagga agctatagat agaattgcag agcttacttt taatatgaat cttgagagtg
     1981 aaaatcttgg tgccagaaga cttcacggtg ttatggaaat agtgcttgca gatctttttt
     2041 ttgaagtgcc tggcagtaag ttgaaaaaat ttgaaataaa cttggactat gttaataaaa
     2101 aaatacaaat taacgaacaa aaagatttga attattatat aatttagtta aaagcaattt
     2161 taaatgaggg aggtttccat atgttacgca gcagcaacga tgttacgcag cagggcagtc
     2221 gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc
     2281 ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg
     2341 tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta
     2401 agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt
     2461 acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct
     2521 ccggcgagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg
     2581 ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag
     2641 tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa
     2701 gtaccgccac ctaacaattc gttcaagccg agatcggctt acgcgtccaa aatcccttaa
     2761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2941 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcacta
     3481 gatccccggc aagttcatcc ttagggtttt catatctctt aaaaagtcaa tttctgcttg
     3541 attaatttct tgtggattat tgtttttgcg gtttttcatt ttttatctcc ttgagtattg
     3601 aattttttct tatatataaa atatagcaaa aactattttt gccaactttt ttacaaaaaa
     3661 ttttttatta aaacacttag ggctttgcta aattctcttt taaaagaact tagtaaagcc
     3721 ctatggattt aagaactgat tatacttacg tagtaaaaat actacagata ttaatagtat
     3781 agaccatatc aaaatcataa tcaatttata tttatctaaa attgtggtcc atacaaaata
     3841 caaaaaaatt aggtttttct aaattctatt ttaaaagaac ttagtaacac ctaatctcta
     3901 aattacttac aaaattctaa aattagattt ttagttcttc atatttcttt aaaagttttt
     3961 taagaaaatc tttttgattt tcaaaaattt catgcatcat aaaactagta aatttggatt
     4021 tttttttgta aaaatcataa ctttcttggg ttttaagttg aaatcttaat ggttttattg
     4081 gattttgcct agattttttt aacgccggac tttctttatc ttttataaaa ttataagaat
     4141 ttttaatacc attttctatt aaataatctt cttttataag cccctcttgc agtgcagttg
     4201 ctaatcttaa atacttatac gcttgagttt ttgctacatc aaagtctttt ataaaatcaa
     4261 gaaaagtttt ataaccatca aatttataat attgattatc ttttatttct tttaaaattt
     4321 taatattttc caccctgtgg aatatttctt ttttgaaatt gtcatttaac ttttccttta
     4381 gtgtcttgta ataagaaatt ttactttcat taatcattaa attttgttca acataagtat
     4441 ctatcttttt tgttaaaact ttaacaacac gttttttttc cataaaaatt ttcctttatt
     4501 tttgcaacat tttccttaat tgatataatt gttactattt tagatttttt aatatccact
     4561 taagaattaa tccctcacta gaatattttt tattatagaa ttcatataca tgtttttcta
     4621 aatctataaa tttatcaaca attgctttat tatagggtgt atctatcttt tcttttctta
     4681 ataatcgtct catattcttt aaataacaac aaacactgcc ggttttaaac ctaaattcta
     4741 tgtaatatat ttttgaaaaa gtatatacat tttcaacccc gtttatttca tactttacaa
     4801 aaacattatt tattggtttt ctataaccat agtaaatgcc taaaaatttg tccccgtctt
     4861 tcataggata tagataaagt ttttgaatct ctgatttatc taataatttt tcaaaaaaaa
     4921 ttatacaccg cttttgtttg ttactaacta ctctatgcaa tgcctttaat atttttgtgt
     4981 gatatatatt tttatcacct tttttttctt tcagcaggaa gatactttct tttggaaatt
     5041 taattttttg ttcatttatt cttgcagcct taagtccgat tttcatattt aatccttata
     5101 tagcctttct atttaattct tttaaactca ataaactttt ttggtttttc attagttcta
     5161 ataattcgca ataatatgtg caaaacactt tattgtattc tattttcttt tggttattca
     5221 aatattcttt tataattggc tttagaagtt caatatttac ttcttttttt agttgttcaa
     5281 taaggatatt gaaaatgttt gtttttaaat cttgataatt ttgttgagaa ttttcttttt
     5341 ttctttcaat tgacttctct aatttacgct ttatattgtt taaatcgcta tatttatgac
     5401 tttcaataat aaaatgaggc ttaaatttgt aattttcgta tattttttgg aaacttgttt
     5461 ctaattgttt agaattatat tcattttttt caaattcttt ttgagtgttg catagaattt
     5521 cctttaattt ttcttgtttt tctttcatac aagatttagt aaaatgtata tttttatttt
     5581 ttagtaattt aatttctatt atttttatga ttttaagtat tttgattata ttatctttat
     5641 caatatttam aktgaataaa attgtaagaa aaattgaaag aattttttta cataaaaagt
     5701 tacatttatt gaagtaa
//
