LOCUS       sequence-311197-        5653 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 5653)
  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 5653)
  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_P0026-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5653
                     /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"
     misc_RNA        558..633
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(651..668)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(651..667)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(660..682)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1234..1767
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1870..2458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2359..2378
                     /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 gagctctatt
      421 ctccattctt ttaaaattat tatccttgtt tattatagtt taaaataaat ttaattacaa
      481 tttgaaattt ggtcgtttga ctttccttat attgacaatc taagtataat attaaggaaa
      541 atttaaattt ctgacttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      601 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gaagcttggc actggccgtc
      661 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      721 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      781 cagttgcgca gcctgaatgg cgaatggcgc ggccgcccta ggttaatacc cgagcttcaa
      841 ggaagatttc ctattaaggt tgaacttaag agcttaagca tagacgattt gaaaaaaatt
      901 ttaaaacaaa caaaaaattc tttaataaaa caatatgttg cgatgtttaa ggtttatgat
      961 ttagatttaa agtttagtga ggaagctata gatagaattg cagagcttac ttttaatatg
     1021 aatcttgaga gtgaaaatct tggtgccaga agacttcacg gtgttatgga aatagtgctt
     1081 gcagatcttt tttttgaagt gcctggcagt aagttgaaaa aatttgaaat aaacttggac
     1141 tatgttaata aaaaaataca aattaacgaa caaaaagatt tgaattatta tataatttag
     1201 ttaaaagcaa ttttaaatga gggaggtttc catatgttac gcagcagcaa cgatgttacg
     1261 cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca
     1321 tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt
     1381 gagttcggag acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac
     1441 ttgctccgta gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc
     1501 gctctcgcgg cttacgttct gcccaagttt gagcagccgc gtagtgagat ctatatctat
     1561 gatctcgcag tctccggcga gcaccggagg cagggcattg ccaccgcgct catcaatctc
     1621 ctcaagcatg aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt
     1681 gacgatcccg cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt
     1741 gatatcgacc caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttacgcgtc
     1801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2521 ctttcctgca ctagatcccc ggcaagttca tccttagggt tttcatatct cttaaaaagt
     2581 caatttctgc ttgattaatt tcttgtggat tattgttttt gcggtttttc attttttatc
     2641 tccttgagta ttgaattttt tcttatatat aaaatatagc aaaaactatt tttgccaact
     2701 tttttacaaa aaatttttta ttaaaacact tagggctttg ctaaattctc ttttaaaaga
     2761 acttagtaaa gccctatgga tttaagaact gattatactt acgtagtaaa aatactacag
     2821 atattaatag tatagaccat atcaaaatca taatcaattt atatttatct aaaattgtgg
     2881 tccatacaaa atacaaaaaa attaggtttt tctaaattct attttaaaag aacttagtaa
     2941 cacctaatct ctaaattact tacaaaattc taaaattaga tttttagttc ttcatatttc
     3001 tttaaaagtt ttttaagaaa atctttttga ttttcaaaaa tttcatgcat cataaaacta
     3061 gtaaatttgg attttttttt gtaaaaatca taactttctt gggttttaag ttgaaatctt
     3121 aatggtttta ttggattttg cctagatttt tttaacgccg gactttcttt atcttttata
     3181 aaattataag aatttttaat accattttct attaaataat cttcttttat aagcccctct
     3241 tgcagtgcag ttgctaatct taaatactta tacgcttgag tttttgctac atcaaagtct
     3301 tttataaaat caagaaaagt tttataacca tcaaatttat aatattgatt atcttttatt
     3361 tcttttaaaa ttttaatatt ttccaccctg tggaatattt cttttttgaa attgtcattt
     3421 aacttttcct ttagtgtctt gtaataagaa attttacttt cattaatcat taaattttgt
     3481 tcaacataag tatctatctt ttttgttaaa actttaacaa cacgtttttt ttccataaaa
     3541 attttccttt atttttgcaa cattttcctt aattgatata attgttacta ttttagattt
     3601 tttaatatcc acttaagaat taatccctca ctagaatatt ttttattata gaattcatat
     3661 acatgttttt ctaaatctat aaatttatca acaattgctt tattataggg tgtatctatc
     3721 ttttcttttc ttaataatcg tctcatattc tttaaataac aacaaacact gccggtttta
     3781 aacctaaatt ctatgtaata tatttttgaa aaagtatata cattttcaac cccgtttatt
     3841 tcatacttta caaaaacatt atttattggt tttctataac catagtaaat gcctaaaaat
     3901 ttgtccccgt ctttcatagg atatagataa agtttttgaa tctctgattt atctaataat
     3961 ttttcaaaaa aaattataca ccgcttttgt ttgttactaa ctactctatg caatgccttt
     4021 aatatttttg tgtgatatat atttttatca cctttttttt ctttcagcag gaagatactt
     4081 tcttttggaa atttaatttt ttgttcattt attcttgcag ccttaagtcc gattttcata
     4141 tttaatcctt atatagcctt tctatttaat tcttttaaac tcaataaact tttttggttt
     4201 ttcattagtt ctaataattc gcaataatat gtgcaaaaca ctttattgta ttctattttc
     4261 ttttggttat tcaaatattc ttttataatt ggctttagaa gttcaatatt tacttctttt
     4321 tttagttgtt caataaggat attgaaaatg tttgttttta aatcttgata attttgttga
     4381 gaattttctt tttttctttc aattgacttc tctaatttac gctttatatt gtttaaatcg
     4441 ctatatttat gactttcaat aataaaatga ggcttaaatt tgtaattttc gtatattttt
     4501 tggaaacttg tttctaattg tttagaatta tattcatttt tttcaaattc tttttgagtg
     4561 ttgcatagaa tttcctttaa tttttcttgt ttttctttca tacaagattt agtaaaatgt
     4621 atatttttat tttttagtaa tttaatttct attattttta tgattttaag tattttgatt
     4681 atattatctt tatcaatatt taaattgaat aaaattgtaa gaaaaattga aagaattttt
     4741 ttacataaaa agttacattt attgaagtaa tttcttactt gatacttttc tatttcatta
     4801 atctttcttt cttcttttat attattttta ttatttaaac actcctttaa atctacactc
     4861 ccatttttag taaatctatc tttaaggccc cttgaaactc tattttgaaa tcttaaatct
     4921 tttttttctt gaaagtattt atagattttt tggtaacatt ctttttttga aaaattaagt
     4981 tgatagtata tttcggtccc caaatttacc cctaagtgtt tgtgataatt agttgtgact
     5041 ttaaattctt tttctaattt gtaaaggtaa ttttgcagtg tttttagttt aacgggtgtt
     5101 tgactatttc ttttaaggtt ttcattaaag taatagagta tgtttttttg tgtgtatttc
     5161 ttatatgttt tatttatgta cgctagtgtt gagataagga ctactaattt atactgtaat
     5221 ttattgtagc aagttgtggt tttactggtt ttgtttatat tagataagtt ttccatttat
     5281 agctccttga ataccggatt ttttatctat aaagtatata tatcacaatt ttgatttaaa
     5341 agtaaattct ttttttaaaa atacaaaaat tttttataaa ttatatagac catgatattg
     5401 tattaatact aattgatttt aatttaaggt tgaactaaaa taaacatagt tttgtagaaa
     5461 ataatttttc attattttgc tacttgatat tggatcgtaa aaatattagg gctttacaaa
     5521 gttcttttaa aagagaattt agtaaagccc taagtctttt aataaaaaat tttttgtaaa
     5581 aaagttggca aaaatagttt ttgctatata tttatatata agaaaaaatt caatactcaa
     5641 ggagataaaa aat
//
