LOCUS       sequence-311205-        6209 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 6209)
  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 6209)
  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_P0526-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1260..1281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1296..1301,1302..1319,1320..1326)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1334..1350
                     /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     1339..1361
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1358..1374
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1358..1374
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        2092..2167
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2185..2202)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2185..2201)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2194..2216)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2768..3301
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      3404..3992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3893..3912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgattatatt atctttatca atatttaaat tgaataaaat tgtaagaaaa attgaaagaa
       61 tttttttaca taaaaagtta catttattga agtaatttct tacttgatac ttttctattt
      121 cattaatctt tctttcttct tttatattat ttttattatt taaacactcc tttaaatcta
      181 cactcccatt tttagtaaat ctatctttaa ggccccttga aactctattt tgaaatctta
      241 aatctttttt ttcttgaaag tatttataga ttttttggta acattctttt tttgaaaaat
      301 taagttgata gtatatttcg gtccccaaat ttacccctaa gtgtttgtga taattagttg
      361 tgactttaaa ttctttttct aatttgtaaa ggtaattttg cagtgttttt agtttaacgg
      421 gtgtttgact atttctttta aggttttcat taaagtaata gagtatgttt ttttgtgtgt
      481 atttcttata tgttttattt atgtacgcta gtgttgagat aaggactact aatttatact
      541 gtaatttatt gtagcaagtt gtggttttac tggttttgtt tatattagat aagttttcca
      601 tttatagctc cttgaatacc ggatttttta tctataaagt atatatatca caattttgat
      661 ttaaaagtaa attctttttt taaaaataca aaaatttttt ataaattata tagaccatga
      721 tattgtatta atactaattg attttaattt aaggttgaac taaaataaac atagttttgt
      781 agaaaataat ttttcattat tttgctactt gatattggat cgtaaaaata ttagggcttt
      841 acaaagttct tttaaaagag aatttagtaa agccctaagt cttttaataa aaaatttttt
      901 gtaaaaaagt tggcaaaaat agtttttgct atatatttat atataagaaa aaattcaata
      961 ctcaaggaga taaaaaatgg aaaacttaaa caataatcca caaaaaatta atcaaataaa
     1021 taagcaaact aattttattg gttttgaaaa ttttatagaa cttgataatc aagttaatca
     1081 aactaaagaa aaaagtaaat taagtaagat aggtaagaaa ttacccggaa taagcagtca
     1141 agtctagtgc ggccgcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1201 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     1261 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     1321 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
     1381 acgaattcga gctccttgtt ttcaatgata ggttttttag gtagagactt gtaagggttt
     1441 gttcctggaa tataattttt aggaattttc atatatttta ctcctattta actagctaac
     1501 ctataattct attatttttt ttgtttttat gcaatcaata ttgatatttt gattttttat
     1561 gattagaatc atcatggaag agccaaagtc aacacttata gacaaatatg atctttctaa
     1621 ccttcacgaa atagcatcag ggggagcacc tttgtctaag gaagtaggag aggcagttgc
     1681 aaagaggttt catttacccg gaattaggca aggatatggg ttaacagaaa ccacatctgc
     1741 aattttaatt acacctgaag gagatgataa accaggagct gttgggaagg ttgttccttt
     1801 ttttgaggct aaagttgttg atttagatac tggaaagact ttaggagtta atcaaagagg
     1861 cgagttatgt gtaagaggac ctatgataat gtcaggatac gtaaataatc cagaagctac
     1921 aaatgcattg atagataaag atggttggtt acactcaggc gatatagctt attgggacga
     1981 agatgaacac ttttttatag ttgatagatt gaaatctttg attaagtata aaggttatca
     2041 agtagctcca gcagaattgg aaagtatttt attgcaacac ccagctcttc cgttttagag
     2101 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     2161 tcggtgcttt ttttgaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2221 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2281 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2341 gcgcggccgc cctaggttaa tacccgagct tcaaggaaga tttcctatta aggttgaact
     2401 taagagctta agcatagacg atttgaaaaa aattttaaaa caaacaaaaa attctttaat
     2461 aaaacaatat gttgcgatgt ttaaggttta tgatttagat ttaaagttta gtgaggaagc
     2521 tatagataga attgcagagc ttacttttaa tatgaatctt gagagtgaaa atcttggtgc
     2581 cagaagactt cacggtgtta tggaaatagt gcttgcagat cttttttttg aagtgcctgg
     2641 cagtaagttg aaaaaatttg aaataaactt ggactatgtt aataaaaaaa tacaaattaa
     2701 cgaacaaaaa gatttgaatt attatataat ttagttaaaa gcaattttaa atgagggagg
     2761 tttccatatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     2821 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     2881 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     2941 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     3001 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa
     3061 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gcgagcaccg
     3121 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     3181 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     3241 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     3301 acaattcgtt caagccgaga tcggcttacg cgtccaaaat cccttaacgt gagttttcgt
     3361 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3421 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3481 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3541 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3601 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3661 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3721 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3781 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3841 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3901 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3961 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcactagat ccccggcaag
     4081 ttcatcctta gggttttcat atctcttaaa aagtcaattt ctgcttgatt aatttcttgt
     4141 ggattattgt ttttgcggtt tttcattttt tatctccttg agtattgaat tttttcttat
     4201 atataaaata tagcaaaaac tatttttgcc aactttttta caaaaaattt tttattaaaa
     4261 cacttagggc tttgctaaat tctcttttaa aagaacttag taaagcccta tggatttaag
     4321 aactgattat acttacgtag taaaaatact acagatatta atagtataga ccatatcaaa
     4381 atcataatca atttatattt atctaaaatt gtggtccata caaaatacaa aaaaattagg
     4441 tttttctaaa ttctatttta aaagaactta gtaacaccta atctctaaat tacttacaaa
     4501 attctaaaat tagattttta gttcttcata tttctttaaa agttttttaa gaaaatcttt
     4561 ttgattttca aaaatttcat gcatcataaa actagtaaat ttggattttt ttttgtaaaa
     4621 atcataactt tcttgggttt taagttgaaa tcttaatggt tttattggat tttgcctaga
     4681 tttttttaac gccggacttt ctttatcttt tataaaatta taagaatttt taataccatt
     4741 ttctattaaa taatcttctt ttataagccc ctcttgcagt gcagttgcta atcttaaata
     4801 cttatacgct tgagtttttg ctacatcaaa gtcttttata aaatcaagaa aagttttata
     4861 accatcaaat ttataatatt gattatcttt tatttctttt aaaattttaa tattttccac
     4921 cctgtggaat atttcttttt tgaaattgtc atttaacttt tcctttagtg tcttgtaata
     4981 agaaatttta ctttcattaa tcattaaatt ttgttcaaca taagtatcta tcttttttgt
     5041 taaaacttta acaacacgtt ttttttccat aaaaattttc ctttattttt gcaacatttt
     5101 ccttaattga tataattgtt actattttag attttttaat atccacttaa gaattaatcc
     5161 ctcactagaa tattttttat tatagaattc atatacatgt ttttctaaat ctataaattt
     5221 atcaacaatt gctttattat agggtgtatc tatcttttct tttcttaata atcgtctcat
     5281 attctttaaa taacaacaaa cactgccggt tttaaaccta aattctatgt aatatatttt
     5341 tgaaaaagta tatacatttt caaccccgtt tatttcatac tttacaaaaa cattatttat
     5401 tggttttcta taaccatagt aaatgcctaa aaatttgtcc ccgtctttca taggatatag
     5461 ataaagtttt tgaatctctg atttatctaa taatttttca aaaaaaatta tacaccgctt
     5521 ttgtttgtta ctaactactc tatgcaatgc ctttaatatt tttgtgtgat atatattttt
     5581 atcacctttt ttttctttca gcaggaagat actttctttt ggaaatttaa ttttttgttc
     5641 atttattctt gcagccttaa gtccgatttt catatttaat ccttatatag cctttctatt
     5701 taattctttt aaactcaata aacttttttg gtttttcatt agttctaata attcgcaata
     5761 atatgtgcaa aacactttat tgtattctat tttcttttgg ttattcaaat attcttttat
     5821 aattggcttt agaagttcaa tatttacttc tttttttagt tgttcaataa ggatattgaa
     5881 aatgtttgtt tttaaatctt gataattttg ttgagaattt tctttttttc tttcaattga
     5941 cttctctaat ttacgcttta tattgtttaa atcgctatat ttatgacttt caataataaa
     6001 atgaggctta aatttgtaat tttcgtatat tttttggaaa cttgtttcta attgtttaga
     6061 attatattca tttttttcaa attctttttg agtgttgcat agaatttcct ttaatttttc
     6121 ttgtttttct ttcatacaag atttagtaaa atgtatattt ttatttttta gtaatttaat
     6181 ttctattatt tttatgattt taagtattt
//
