LOCUS       sequence-311488-        5693 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 5693)
  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 5693)
  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_P0826S-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..35)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(18..34)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(27..49)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             601..1134
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1237..1825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1726..1745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5302..5323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5338..5343,5344..5361,5362..5368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5376..5392
                     /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     5381..5403
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5400..5416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5400..5416
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        5618..5693
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tttttttgaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
       61 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      121 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcggc
      181 cgccctaggt taatacccga gcttcaagga agatttccta ttaaggttga acttaagagc
      241 ttaagcatag acgatttgaa aaaaatttta aaacaaacaa aaaattcttt aataaaacaa
      301 tatgttgcga tgtttaaggt ttatgattta gatttaaagt ttagtgagga agctatagat
      361 agaattgcag agcttacttt taatatgaat cttgagagtg aaaatcttgg tgccagaaga
      421 cttcacggtg ttatggaaat agtgcttgca gatctttttt ttgaagtgcc tggcagtaag
      481 ttgaaaaaat ttgaaataaa cttggactat gttaataaaa aaatacaaat taacgaacaa
      541 aaagatttga attattatat aatttagtta aaagcaattt taaatgaggg aggtttccat
      601 atgttacgca gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaggt
      661 ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt caaatccatg
      721 cgggctgctc ttgatctttt cggtcgtgag ttcggagacg tagccaccta ctcccaacat
      781 cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat cgcgcttgct
      841 gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc caagtttgag
      901 cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggcgagca ccggaggcag
      961 ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct tggtgcttat
     1021 gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta tacaaagttg
     1081 ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac ctaacaattc
     1141 gttcaagccg agatcggctt acgcgtccaa aatcccttaa cgtgagtttt cgttccactg
     1201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1381 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1861 cttttgctgg ccttttgctc acatgttctt tcctgcacta gatccccggc aagttcatcc
     1921 ttagggtttt catatctctt aaaaagtcaa tttctgcttg attaatttct tgtggattat
     1981 tgtttttgcg gtttttcatt ttttatctcc ttgagtattg aattttttct tatatataaa
     2041 atatagcaaa aactattttt gccaactttt ttacaaaaaa ttttttatta aaacacttag
     2101 ggctttgcta aattctcttt taaaagaact tagtaaagcc ctatggattt aagaactgat
     2161 tatacttacg tagtaaaaat actacagata ttaatagtat agaccatatc aaaatcataa
     2221 tcaatttata tttatctaaa attgtggtcc atacaaaata caaaaaaatt aggtttttct
     2281 aaattctatt ttaaaagaac ttagtaacac ctaatctcta aattacttac aaaattctaa
     2341 aattagattt ttagttcttc atatttcttt aaaagttttt taagaaaatc tttttgattt
     2401 tcaaaaattt catgcatcat aaaactagta aatttggatt tttttttgta aaaatcataa
     2461 ctttcttggg ttttaagttg aaatcttaat ggttttattg gattttgcct agattttttt
     2521 aacgccggac tttctttatc ttttataaaa ttataagaat ttttaatacc attttctatt
     2581 aaataatctt cttttataag cccctcttgc agtgcagttg ctaatcttaa atacttatac
     2641 gcttgagttt ttgctacatc aaagtctttt ataaaatcaa gaaaagtttt ataaccatca
     2701 aatttataat attgattatc ttttatttct tttaaaattt taatattttc caccctgtgg
     2761 aatatttctt ttttgaaatt gtcatttaac ttttccttta gtgtcttgta ataagaaatt
     2821 ttactttcat taatcattaa attttgttca acataagtat ctatcttttt tgttaaaact
     2881 ttaacaacac gttttttttc cataaaaatt ttcctttatt tttgcaacat tttccttaat
     2941 tgatataatt gttactattt tagatttttt aatatccact taagaattaa tccctcacta
     3001 gaatattttt tattatagaa ttcatataca tgtttttcta aatctataaa tttatcaaca
     3061 attgctttat tatagggtgt atctatcttt tcttttctta ataatcgtct catattcttt
     3121 aaataacaac aaacactgcc ggttttaaac ctaaattcta tgtaatatat ttttgaaaaa
     3181 gtatatacat tttcaacccc gtttatttca tactttacaa aaacattatt tattggtttt
     3241 ctataaccat agtaaatgcc taaaaatttg tccccgtctt tcataggata tagataaagt
     3301 ttttgaatct ctgatttatc taataatttt tcaaaaaaaa ttatacaccg cttttgtttg
     3361 ttactaacta ctctatgcaa tgcctttaat atttttgtgt gatatatatt tttatcacct
     3421 tttttttctt tcagcaggaa gatactttct tttggaaatt taattttttg ttcatttatt
     3481 cttgcagcct taagtccgat tttcatattt aatccttata tagcctttct atttaattct
     3541 tttaaactca ataaactttt ttggtttttc attagttcta ataattcgca ataatatgtg
     3601 caaaacactt tattgtattc tattttcttt tggttattca aatattcttt tataattggc
     3661 tttagaagtt caatatttac ttcttttttt agttgttcaa taaggatatt gaaaatgttt
     3721 gtttttaaat cttgataatt ttgttgagaa ttttcttttt ttctttcaat tgacttctct
     3781 aatttacgct ttatattgtt taaatcgcta tatttatgac tttcaataat aaaatgaggc
     3841 ttaaatttgt aattttcgta tattttttgg aaacttgttt ctaattgttt agaattatat
     3901 tcattttttt caaattcttt ttgagtgttg catagaattt cctttaattt ttcttgtttt
     3961 tctttcatac aagatttagt aaaatgtata tttttatttt ttagtaattt aatttctatt
     4021 atttttatga ttttaagtat tttgattata ttatctttat caatatttaa attgaataaa
     4081 attgtaagaa aaattgaaag aattttttta cataaaaagt tacatttatt gaagtaattt
     4141 cttacttgat acttttctat ttcattaatc tttctttctt cttttatatt atttttatta
     4201 tttaaacact cctttaaatc tacactccca tttttagtaa atctatcttt aaggcccctt
     4261 gaaactctat tttgaaatct taaatctttt ttttcttgaa agtatttata gattttttgg
     4321 taacattctt tttttgaaaa attaagttga tagtatattt cggtccccaa atttacccct
     4381 aagtgtttgt gataattagt tgtgacttta aattcttttt ctaatttgta aaggtaattt
     4441 tgcagtgttt ttagtttaac gggtgtttga ctatttcttt taaggttttc attaaagtaa
     4501 tagagtatgt ttttttgtgt gtatttctta tatgttttat ttatgtacgc tagtgttgag
     4561 ataaggacta ctaatttata ctgtaattta ttgtagcaag ttgtggtttt actggttttg
     4621 tttatattag ataagttttc catttatagc tccttgaata ccggattttt tatctataaa
     4681 gtatatatat cacaattttg atttaaaagt aaattctttt tttaaaaata caaaaatttt
     4741 ttataaatta tatagaccat gatattgtat taatactaat tgattttaat ttaaggttga
     4801 actaaaataa acatagtttt gtagaaaata atttttcatt attttgctac ttgatattgg
     4861 atcgtaaaaa tattagggct ttacaaagtt cttttaaaag agaatttagt aaagccctaa
     4921 gtcttttaat aaaaaatttt ttgtaaaaaa gttggcaaaa atagtttttg ctatatattt
     4981 atatataaga aaaaattcaa tactcaagga gataaaaaat ggaaaactta aacaataatc
     5041 cacaaaaaat taatcaaata aataagcaaa ctaattttat tggttttgaa aattttatag
     5101 aacttgataa tcaagttaat caaactaaag aaaaaagtaa attaagtaag ataggtaaga
     5161 aattacccgg aataagcagt caagtctagt gcggccgcgc ccaatacgca aaccgcctct
     5221 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5281 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5341 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5401 aggaaacagc tatgacatga ttacgaattc gagctcggca atagaagaat ctatagaaag
     5461 cataaaaagc cttatagaaa acaaagaata catacaaaag ttacatttta taaaaaaatt
     5521 aataaataag gtttacaaaa aattaaatta ctttttttaa aaactaaact ttgaaagcct
     5581 tgttataata taaaatgaaa atttaaattt ctgacttgtt ttagagctag aaatagcaag
     5641 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgc
//
