LOCUS       sequence-318251-        6188 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 6188)
  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 6188)
  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-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        812..887
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(905..922)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(905..921)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(914..936)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1488..2021
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2124..2712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2613..2632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattcg agctcggcaa tagaagaatc tatagaaagc ataaaaagcc ttatagaaaa
      181 caaagaatac atacaaaagt tacattttat aaaaaaatta ataaataagg tttacaaaaa
      241 attaaattac tttttttaaa aactaaactt tgaaagcctt gttataatat aaaatggaag
      301 agccaaagtc aacacttata gacaaatatg atctttctaa ccttcacgaa atagcatcag
      361 ggggagcacc tttgtctaag gaagtaggag aggcagttgc aaagaggttt catttacccg
      421 gaattaggca aggatatggg ttaacagaaa ccacatctgc aattttaatt acacctgaag
      481 gagatgataa accaggagct gttgggaagg ttgttccttt ttttgaggct aaagttgttg
      541 atttagatac tggaaagact ttaggagtta atcaaagagg cgagttatgt gtaagaggac
      601 ctatgataat gtcaggatac gtaaataatc cagaagctac aaatgcattg atagataaag
      661 atggttggtt acactcaggc gatatagctt attgggacga agatgaacac ttttttatag
      721 ttgatagatt gaaatctttg attaagtata aaggttatca agtagctcca gcagaattgg
      781 aaagtatttt attgcaacac ccagctcttc cgttttagag ctagaaatag caagttaaaa
      841 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgaagct
      901 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      961 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1021 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcggccgc cctaggttaa
     1081 tacccgagct tcaaggaaga tttcctatta aggttgaact taagagctta agcatagacg
     1141 atttgaaaaa aattttaaaa caaacaaaaa attctttaat aaaacaatat gttgcgatgt
     1201 ttaaggttta tgatttagat ttaaagttta gtgaggaagc tatagataga attgcagagc
     1261 ttacttttaa tatgaatctt gagagtgaaa atcttggtgc cagaagactt cacggtgtta
     1321 tggaaatagt gcttgcagat cttttttttg aagtgcctgg cagtaagttg aaaaaatttg
     1381 aaataaactt ggactatgtt aataaaaaaa tacaaattaa cgaacaaaaa gatttgaatt
     1441 attatataat ttagttaaaa gcaattttaa atgagggagg tttccatatg ttacgcagca
     1501 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg
     1561 gcatcattcg cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg
     1621 atcttttcgg tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg
     1681 attacctcgg gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag
     1741 aagcggttgt tggcgctctc gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg
     1801 agatctatat ctatgatctc gcagtctccg gcgagcaccg gaggcagggc attgccaccg
     1861 cgctcatcaa tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc
     1921 aagcagatta cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag
     1981 aagtgatgca ctttgatatc gacccaagta ccgccaccta acaattcgtt caagccgaga
     2041 tcggcttacg cgtccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2701 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2761 tttgctcaca tgttctttcc tgcactagat ccccggcaag ttcatcctta gggttttcat
     2821 atctcttaaa aagtcaattt ctgcttgatt aatttcttgt ggattattgt ttttgcggtt
     2881 tttcattttt tatctccttg agtattgaat tttttcttat atataaaata tagcaaaaac
     2941 tatttttgcc aactttttta caaaaaattt tttattaaaa cacttagggc tttgctaaat
     3001 tctcttttaa aagaacttag taaagcccta tggatttaag aactgattat acttacgtag
     3061 taaaaatact acagatatta atagtataga ccatatcaaa atcataatca atttatattt
     3121 atctaaaatt gtggtccata caaaatacaa aaaaattagg tttttctaaa ttctatttta
     3181 aaagaactta gtaacaccta atctctaaat tacttacaaa attctaaaat tagattttta
     3241 gttcttcata tttctttaaa agttttttaa gaaaatcttt ttgattttca aaaatttcat
     3301 gcatcataaa actagtaaat ttggattttt ttttgtaaaa atcataactt tcttgggttt
     3361 taagttgaaa tcttaatggt tttattggat tttgcctaga tttttttaac gccggacttt
     3421 ctttatcttt tataaaatta taagaatttt taataccatt ttctattaaa taatcttctt
     3481 ttataagccc ctcttgcagt gcagttgcta atcttaaata cttatacgct tgagtttttg
     3541 ctacatcaaa gtcttttata aaatcaagaa aagttttata accatcaaat ttataatatt
     3601 gattatcttt tatttctttt aaaattttaa tattttccac cctgtggaat atttcttttt
     3661 tgaaattgtc atttaacttt tcctttagtg tcttgtaata agaaatttta ctttcattaa
     3721 tcattaaatt ttgttcaaca taagtatcta tcttttttgt taaaacttta acaacacgtt
     3781 ttttttccat aaaaattttc ctttattttt gcaacatttt ccttaattga tataattgtt
     3841 actattttag attttttaat atccacttaa gaattaatcc ctcactagaa tattttttat
     3901 tatagaattc atatacatgt ttttctaaat ctataaattt atcaacaatt gctttattat
     3961 agggtgtatc tatcttttct tttcttaata atcgtctcat attctttaaa taacaacaaa
     4021 cactgccggt tttaaaccta aattctatgt aatatatttt tgaaaaagta tatacatttt
     4081 caaccccgtt tatttcatac tttacaaaaa cattatttat tggttttcta taaccatagt
     4141 aaatgcctaa aaatttgtcc ccgtctttca taggatatag ataaagtttt tgaatctctg
     4201 atttatctaa taatttttca aaaaaaatta tacaccgctt ttgtttgtta ctaactactc
     4261 tatgcaatgc ctttaatatt tttgtgtgat atatattttt atcacctttt ttttctttca
     4321 gcaggaagat actttctttt ggaaatttaa ttttttgttc atttattctt gcagccttaa
     4381 gtccgatttt catatttaat ccttatatag cctttctatt taattctttt aaactcaata
     4441 aacttttttg gtttttcatt agttctaata attcgcaata atatgtgcaa aacactttat
     4501 tgtattctat tttcttttgg ttattcaaat attcttttat aattggcttt agaagttcaa
     4561 tatttacttc tttttttagt tgttcaataa ggatattgaa aatgtttgtt tttaaatctt
     4621 gataattttg ttgagaattt tctttttttc tttcaattga cttctctaat ttacgcttta
     4681 tattgtttaa atcgctatat ttatgacttt caataataaa atgaggctta aatttgtaat
     4741 tttcgtatat tttttggaaa cttgtttcta attgtttaga attatattca tttttttcaa
     4801 attctttttg agtgttgcat agaatttcct ttaatttttc ttgtttttct ttcatacaag
     4861 atttagtaaa atgtatattt ttatttttta gtaatttaat ttctattatt tttatgattt
     4921 taagtatttt gattatatta tctttatcaa tatttaaatt gaataaaatt gtaagaaaaa
     4981 ttgaaagaat ttttttacat aaaaagttac atttattgaa gtaatttctt acttgatact
     5041 tttctatttc attaatcttt ctttcttctt ttatattatt tttattattt aaacactcct
     5101 ttaaatctac actcccattt ttagtaaatc tatctttaag gccccttgaa actctatttt
     5161 gaaatcttaa atcttttttt tcttgaaagt atttatagat tttttggtaa cattcttttt
     5221 ttgaaaaatt aagttgatag tatatttcgg tccccaaatt tacccctaag tgtttgtgat
     5281 aattagttgt gactttaaat tctttttcta atttgtaaag gtaattttgc agtgttttta
     5341 gtttaacggg tgtttgacta tttcttttaa ggttttcatt aaagtaatag agtatgtttt
     5401 tttgtgtgta tttcttatat gttttattta tgtacgctag tgttgagata aggactacta
     5461 atttatactg taatttattg tagcaagttg tggttttact ggttttgttt atattagata
     5521 agttttccat ttatagctcc ttgaataccg gattttttat ctataaagta tatatatcac
     5581 aattttgatt taaaagtaaa ttcttttttt aaaaatacaa aaatttttta taaattatat
     5641 agaccatgat attgtattaa tactaattga ttttaattta aggttgaact aaaataaaca
     5701 tagttttgta gaaaataatt tttcattatt ttgctacttg atattggatc gtaaaaatat
     5761 tagggcttta caaagttctt ttaaaagaga atttagtaaa gccctaagtc ttttaataaa
     5821 aaattttttg taaaaaagtt ggcaaaaata gtttttgcta tatatttata tataagaaaa
     5881 aattcaatac tcaaggagat aaaaaatgga aaacttaaac aataatccac aaaaaattaa
     5941 tcaaataaat aagcaaacta attttattgg ttttgaaaat tttatagaac ttgataatca
     6001 agttaatcaa actaaagaaa aaagtaaatt aagtaagata ggtaagaaat tacccggaat
     6061 aagcagtcaa gtctagtgcg gccgcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6121 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6181 aacgcaat
//
