LOCUS       sequence-311473-        6326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA expression in B. burgdorferi, parental vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  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 6326)
  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-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1171..1192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1207..1212,1213..1230,1231..1237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1245..1261
                     /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     1250..1272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1269..1285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1269..1285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        2120..2195
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2213..2230)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2213..2229)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2222..2244)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2796..3329
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      3432..4020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3921..3940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagtaatttc ttacttgata cttttctatt tcattaatct ttctttcttc ttttatatta
       61 tttttattat ttaaacactc ctttaaatct acactcccat ttttagtaaa tctatcttta
      121 aggccccttg aaactctatt ttgaaatctt aaatcttttt tttcttgaaa gtatttatag
      181 attttttggt aacattcttt ttttgaaaaa ttaagttgat agtatatttc ggtccccaaa
      241 tttaccccta agtgtttgtg ataattagtt gtgactttaa attctttttc taatttgtaa
      301 aggtaatttt gcagtgtttt tagtttaacg ggtgtttgac tatttctttt aaggttttca
      361 ttaaagtaat agagtatgtt tttttgtgtg tatttcttat atgttttatt tatgtacgct
      421 agtgttgaga taaggactac taatttatac tgtaatttat tgtagcaagt tgtggtttta
      481 ctggttttgt ttatattaga taagttttcc atttatagct ccttgaatac cggatttttt
      541 atctataaag tatatatatc acaattttga tttaaaagta aattcttttt ttaaaaatac
      601 aaaaattttt tataaattat atagaccatg atattgtatt aatactaatt gattttaatt
      661 taaggttgaa ctaaaataaa catagttttg tagaaaataa tttttcatta ttttgctact
      721 tgatattgga tcgtaaaaat attagggctt tacaaagttc ttttaaaaga gaatttagta
      781 aagccctaag tcttttaata aaaaattttt tgtaaaaaag ttggcaaaaa tagtttttgc
      841 tatatattta tatataagaa aaaattcaat actcaaggag ataaaaaatg gaaaacttaa
      901 acaataatcc acaaaaaatt aatcaaataa ataagcaaac taattttatt ggttttgaaa
      961 attttataga acttgataat caagttaatc aaactaaaga aaaaagtaaa ttaagtaaga
     1021 taggtaagaa attacccgga ataagcagtc aagtctagtg cggccgcgcc caatacgcaa
     1081 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     1141 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     1201 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     1261 atttcacaca ggaaacagct atgacatgat tacgaattcg agctctgtct gtcgcctctt
     1321 gtggcttccg gaggagttat ttataataaa ataaagaatt acgacttgaa aatttttaat
     1381 caaaaaaata aagttgaaga ttataaaaag aagtacgaag atagagagag aaaagtggaa
     1441 ggtgaactta ataccttgga ttttaccgtt aagcgcatga aagatcaaga aaatacatta
     1501 aaggcttttg attttaatca aagaaataaa taataataat tatttttaat gctattgcta
     1561 tttgcgtttc ttttttttta atttttgtgc tattcttttt aacggaagag ccaaagtcaa
     1621 cacttataga caaatatgat ctttctaacc ttcacgaaat agcatcaggg ggagcacctt
     1681 tgtctaagga agtaggagag gcagttgcaa agaggtttca tttacccgga attaggcaag
     1741 gatatgggtt aacagaaacc acatctgcaa ttttaattac acctgaagga gatgataaac
     1801 caggagctgt tgggaaggtt gttccttttt ttgaggctaa agttgttgat ttagatactg
     1861 gaaagacttt aggagttaat caaagaggcg agttatgtgt aagaggacct atgataatgt
     1921 caggatacgt aaataatcca gaagctacaa atgcattgat agataaagat ggttggttac
     1981 actcaggcga tatagcttat tgggacgaag atgaacactt ttttatagtt gatagattga
     2041 aatctttgat taagtataaa ggttatcaag tagctccagc agaattggaa agtattttat
     2101 tgcaacaccc agctcttccg ttttagagct agaaatagca agttaaaata aggctagtcc
     2161 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaagcttg gcactggccg
     2221 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2281 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2341 aacagttgcg cagcctgaat ggcgaatggc gcggccgccc taggttaata cccgagcttc
     2401 aaggaagatt tcctattaag gttgaactta agagcttaag catagacgat ttgaaaaaaa
     2461 ttttaaaaca aacaaaaaat tctttaataa aacaatatgt tgcgatgttt aaggtttatg
     2521 atttagattt aaagtttagt gaggaagcta tagatagaat tgcagagctt acttttaata
     2581 tgaatcttga gagtgaaaat cttggtgcca gaagacttca cggtgttatg gaaatagtgc
     2641 ttgcagatct tttttttgaa gtgcctggca gtaagttgaa aaaatttgaa ataaacttgg
     2701 actatgttaa taaaaaaata caaattaacg aacaaaaaga tttgaattat tatataattt
     2761 agttaaaagc aattttaaat gagggaggtt tccatatgtt acgcagcagc aacgatgtta
     2821 cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca
     2881 catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc
     2941 gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga
     3001 acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg
     3061 gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct
     3121 atgatctcgc agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc
     3181 tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg
     3241 gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact
     3301 ttgatatcga cccaagtacc gccacctaac aattcgttca agccgagatc ggcttacgcg
     3361 tccaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4081 ttctttcctg cactagatcc ccggcaagtt catccttagg gttttcatat ctcttaaaaa
     4141 gtcaatttct gcttgattaa tttcttgtgg attattgttt ttgcggtttt tcatttttta
     4201 tctccttgag tattgaattt tttcttatat ataaaatata gcaaaaacta tttttgccaa
     4261 cttttttaca aaaaattttt tattaaaaca cttagggctt tgctaaattc tcttttaaaa
     4321 gaacttagta aagccctatg gatttaagaa ctgattatac ttacgtagta aaaatactac
     4381 agatattaat agtatagacc atatcaaaat cataatcaat ttatatttat ctaaaattgt
     4441 ggtccataca aaatacaaaa aaattaggtt tttctaaatt ctattttaaa agaacttagt
     4501 aacacctaat ctctaaatta cttacaaaat tctaaaatta gatttttagt tcttcatatt
     4561 tctttaaaag ttttttaaga aaatcttttt gattttcaaa aatttcatgc atcataaaac
     4621 tagtaaattt ggattttttt ttgtaaaaat cataactttc ttgggtttta agttgaaatc
     4681 ttaatggttt tattggattt tgcctagatt tttttaacgc cggactttct ttatctttta
     4741 taaaattata agaattttta ataccatttt ctattaaata atcttctttt ataagcccct
     4801 cttgcagtgc agttgctaat cttaaatact tatacgcttg agtttttgct acatcaaagt
     4861 cttttataaa atcaagaaaa gttttataac catcaaattt ataatattga ttatctttta
     4921 tttcttttaa aattttaata ttttccaccc tgtggaatat ttcttttttg aaattgtcat
     4981 ttaacttttc ctttagtgtc ttgtaataag aaattttact ttcattaatc attaaatttt
     5041 gttcaacata agtatctatc ttttttgtta aaactttaac aacacgtttt ttttccataa
     5101 aaattttcct ttatttttgc aacattttcc ttaattgata taattgttac tattttagat
     5161 tttttaatat ccacttaaga attaatccct cactagaata ttttttatta tagaattcat
     5221 atacatgttt ttctaaatct ataaatttat caacaattgc tttattatag ggtgtatcta
     5281 tcttttcttt tcttaataat cgtctcatat tctttaaata acaacaaaca ctgccggttt
     5341 taaacctaaa ttctatgtaa tatatttttg aaaaagtata tacattttca accccgttta
     5401 tttcatactt tacaaaaaca ttatttattg gttttctata accatagtaa atgcctaaaa
     5461 atttgtcccc gtctttcata ggatatagat aaagtttttg aatctctgat ttatctaata
     5521 atttttcaaa aaaaattata caccgctttt gtttgttact aactactcta tgcaatgcct
     5581 ttaatatttt tgtgtgatat atatttttat cacctttttt ttctttcagc aggaagatac
     5641 tttcttttgg aaatttaatt ttttgttcat ttattcttgc agccttaagt ccgattttca
     5701 tatttaatcc ttatatagcc tttctattta attcttttaa actcaataaa cttttttggt
     5761 ttttcattag ttctaataat tcgcaataat atgtgcaaaa cactttattg tattctattt
     5821 tcttttggtt attcaaatat tcttttataa ttggctttag aagttcaata tttacttctt
     5881 tttttagttg ttcaataagg atattgaaaa tgtttgtttt taaatcttga taattttgtt
     5941 gagaattttc tttttttctt tcaattgact tctctaattt acgctttata ttgtttaaat
     6001 cgctatattt atgactttca ataataaaat gaggcttaaa tttgtaattt tcgtatattt
     6061 tttggaaact tgtttctaat tgtttagaat tatattcatt tttttcaaat tctttttgag
     6121 tgttgcatag aatttccttt aatttttctt gtttttcttt catacaagat ttagtaaaat
     6181 gtatattttt attttttagt aatttaattt ctattatttt tatgatttta agtattttga
     6241 ttatattatc tttatcaata tttaaattga ataaaattgt aagaaaaatt gaaagaattt
     6301 ttttacataa aaagttacat ttattg
//
