LOCUS       sequence-311091-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rodA gRNA expression in B. burgdorferi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  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 5608)
  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_Psyn-sgRNArodA
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        233..267
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        289..364
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(407..424)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(407..423)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(416..438)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             990..1523
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1626..2214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2115..2134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgacatg attacgaatt cgagctcgaa ttctaaagat ctttgacagc
      241 tagctcagtc ctaggtataa tactagtgtt aatctaccta atataccagt tttagagcta
      301 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      361 gtgctttttt tgaagcttgg gcccgaacaa aaactcaaag cttggcactg gccgtcgttt
      421 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      481 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      541 tgcgcagcct gaatggcgaa tggcgcggcc gccctaggtt aatacccgag cttcaaggaa
      601 gatttcctat taaggttgaa cttaagagct taagcataga cgatttgaaa aaaattttaa
      661 aacaaacaaa aaattcttta ataaaacaat atgttgcgat gtttaaggtt tatgatttag
      721 atttaaagtt tagtgaggaa gctatagata gaattgcaga gcttactttt aatatgaatc
      781 ttgagagtga aaatcttggt gccagaagac ttcacggtgt tatggaaata gtgcttgcag
      841 atcttttttt tgaagtgcct ggcagtaagt tgaaaaaatt tgaaataaac ttggactatg
      901 ttaataaaaa aatacaaatt aacgaacaaa aagatttgaa ttattatata atttagttaa
      961 aagcaatttt aaatgaggga ggtttccata tgttacgcag cagcaacgat gttacgcagc
     1021 agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta
     1081 ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt
     1141 tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc gggaacttgc
     1201 tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt gttggcgctc
     1261 tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat atctatgatc
     1321 tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc aatctcctca
     1381 agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg
     1441 atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata
     1501 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggctta cgcgtccaaa
     1561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2281 cctgcactag atccccggca agttcatcct tagggttttc atatctctta aaaagtcaat
     2341 ttctgcttga ttaatttctt gtggattatt gtttttgcgg tttttcattt tttatctcct
     2401 tgagtattga attttttctt atatataaaa tatagcaaaa actatttttg ccaacttttt
     2461 tacaaaaaat tttttattaa aacacttagg gctttgctaa attctctttt aaaagaactt
     2521 agtaaagccc tatggattta agaactgatt atacttacgt agtaaaaata ctacagatat
     2581 taatagtata gaccatatca aaatcataat caatttatat ttatctaaaa ttgtggtcca
     2641 tacaaaatac aaaaaaatta ggtttttcta aattctattt taaaagaact tagtaacacc
     2701 taatctctaa attacttaca aaattctaaa attagatttt tagttcttca tatttcttta
     2761 aaagtttttt aagaaaatct ttttgatttt caaaaatttc atgcatcata aaactagtaa
     2821 atttggattt ttttttgtaa aaatcataac tttcttgggt tttaagttga aatcttaatg
     2881 gttttattgg attttgccta gattttttta acgccggact ttctttatct tttataaaat
     2941 tataagaatt tttaatacca ttttctatta aataatcttc ttttataagc ccctcttgca
     3001 gtgcagttgc taatcttaaa tacttatacg cttgagtttt tgctacatca aagtctttta
     3061 taaaatcaag aaaagtttta taaccatcaa atttataata ttgattatct tttatttctt
     3121 ttaaaatttt aatattttcc accctgtgga atatttcttt tttgaaattg tcatttaact
     3181 tttcctttag tgtcttgtaa taagaaattt tactttcatt aatcattaaa ttttgttcaa
     3241 cataagtatc tatctttttt gttaaaactt taacaacacg ttttttttcc ataaaaattt
     3301 tcctttattt ttgcaacatt ttccttaatt gatataattg ttactatttt agatttttta
     3361 atatccactt aagaattaat ccctcactag aatatttttt attatagaat tcatatacat
     3421 gtttttctaa atctataaat ttatcaacaa ttgctttatt atagggtgta tctatctttt
     3481 cttttcttaa taatcgtctc atattcttta aataacaaca aacactgccg gttttaaacc
     3541 taaattctat gtaatatatt tttgaaaaag tatatacatt ttcaaccccg tttatttcat
     3601 actttacaaa aacattattt attggttttc tataaccata gtaaatgcct aaaaatttgt
     3661 ccccgtcttt cataggatat agataaagtt tttgaatctc tgatttatct aataattttt
     3721 caaaaaaaat tatacaccgc ttttgtttgt tactaactac tctatgcaat gcctttaata
     3781 tttttgtgtg atatatattt ttatcacctt ttttttcttt cagcaggaag atactttctt
     3841 ttggaaattt aattttttgt tcatttattc ttgcagcctt aagtccgatt ttcatattta
     3901 atccttatat agcctttcta tttaattctt ttaaactcaa taaacttttt tggtttttca
     3961 ttagttctaa taattcgcaa taatatgtgc aaaacacttt attgtattct attttctttt
     4021 ggttattcaa atattctttt ataattggct ttagaagttc aatatttact tcttttttta
     4081 gttgttcaat aaggatattg aaaatgtttg tttttaaatc ttgataattt tgttgagaat
     4141 tttctttttt tctttcaatt gacttctcta atttacgctt tatattgttt aaatcgctat
     4201 atttatgact ttcaataata aaatgaggct taaatttgta attttcgtat attttttgga
     4261 aacttgtttc taattgttta gaattatatt catttttttc aaattctttt tgagtgttgc
     4321 atagaatttc ctttaatttt tcttgttttt ctttcataca agatttagta aaatgtatat
     4381 ttttattttt tagtaattta atttctatta tttttatgat tttaagtatt ttgattatat
     4441 tatctttatc aatatttaaa ttgaataaaa ttgtaagaaa aattgaaaga atttttttac
     4501 ataaaaagtt acatttattg aagtaatttc ttacttgata cttttctatt tcattaatct
     4561 ttctttcttc ttttatatta tttttattat ttaaacactc ctttaaatct acactcccat
     4621 ttttagtaaa tctatcttta aggccccttg aaactctatt ttgaaatctt aaatcttttt
     4681 tttcttgaaa gtatttatag attttttggt aacattcttt ttttgaaaaa ttaagttgat
     4741 agtatatttc ggtccccaaa tttaccccta agtgtttgtg ataattagtt gtgactttaa
     4801 attctttttc taatttgtaa aggtaatttt gcagtgtttt tagtttaacg ggtgtttgac
     4861 tatttctttt aaggttttca ttaaagtaat agagtatgtt tttttgtgtg tatttcttat
     4921 atgttttatt tatgtacgct agtgttgaga taaggactac taatttatac tgtaatttat
     4981 tgtagcaagt tgtggtttta ctggttttgt ttatattaga taagttttcc atttatagct
     5041 ccttgaatac cggatttttt atctataaag tatatatatc acaattttga tttaaaagta
     5101 aattcttttt ttaaaaatac aaaaattttt tataaattat atagaccatg atattgtatt
     5161 aatactaatt gattttaatt taaggttgaa ctaaaataaa catagttttg tagaaaataa
     5221 tttttcatta ttttgctact tgatattgga tcgtaaaaat attagggctt tacaaagttc
     5281 ttttaaaaga gaatttagta aagccctaag tcttttaata aaaaattttt tgtaaaaaag
     5341 ttggcaaaaa tagtttttgc tatatattta tatataagaa aaaattcaat actcaaggag
     5401 ataaaaaatg gaaaacttaa acaataatcc acaaaaaatt aatcaaataa ataagcaaac
     5461 taattttatt ggttttgaaa attttataga acttgataat caagttaatc aaactaaaga
     5521 aaaaagtaaa ttaagtaaga taggtaagaa attacccgga ataagcagtc aagtctagtg
     5581 cggccgcgcc caatacgcaa accgcctc
//
