LOCUS       sequence-311491-        6212 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 6212)
  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 6212)
  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_P0031-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        628..703
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(721..738)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(721..737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(730..752)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1304..1837
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1940..2528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2429..2448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6005..6026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6041..6046,6047..6064,6065..6071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6079..6095
                     /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     6084..6106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6103..6119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6103..6119
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gctttaattt aacttcaaaa agctcaacat ttgaattctc gtcaacctct ttcatctttt
       61 aaagtataga aaagtattta aataagtgtc aatattttgt attatttaat tggaagagcc
      121 aaagtcaaca cttatagaca aatatgatct ttctaacctt cacgaaatag catcaggggg
      181 agcacctttg tctaaggaag taggagaggc agttgcaaag aggtttcatt tacccggaat
      241 taggcaagga tatgggttaa cagaaaccac atctgcaatt ttaattacac ctgaaggaga
      301 tgataaacca ggagctgttg ggaaggttgt tccttttttt gaggctaaag ttgttgattt
      361 agatactgga aagactttag gagttaatca aagaggcgag ttatgtgtaa gaggacctat
      421 gataatgtca ggatacgtaa ataatccaga agctacaaat gcattgatag ataaagatgg
      481 ttggttacac tcaggcgata tagcttattg ggacgaagat gaacactttt ttatagttga
      541 tagattgaaa tctttgatta agtataaagg ttatcaagta gctccagcag aattggaaag
      601 tattttattg caacacccag ctcttccgtt ttagagctag aaatagcaag ttaaaataag
      661 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gaagcttggc
      721 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      781 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      841 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ggccgcccta ggttaatacc
      901 cgagcttcaa ggaagatttc ctattaaggt tgaacttaag agcttaagca tagacgattt
      961 gaaaaaaatt ttaaaacaaa caaaaaattc tttaataaaa caatatgttg cgatgtttaa
     1021 ggtttatgat ttagatttaa agtttagtga ggaagctata gatagaattg cagagcttac
     1081 ttttaatatg aatcttgaga gtgaaaatct tggtgccaga agacttcacg gtgttatgga
     1141 aatagtgctt gcagatcttt tttttgaagt gcctggcagt aagttgaaaa aatttgaaat
     1201 aaacttggac tatgttaata aaaaaataca aattaacgaa caaaaagatt tgaattatta
     1261 tataatttag ttaaaagcaa ttttaaatga gggaggtttc catatgttac gcagcagcaa
     1321 cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat
     1381 cattcgcaca tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct
     1441 tttcggtcgt gagttcggag acgtagccac ctactcccaa catcagccgg actccgatta
     1501 cctcgggaac ttgctccgta gtaagacatt catcgcgctt gctgccttcg accaagaagc
     1561 ggttgttggc gctctcgcgg cttacgttct gcccaagttt gagcagccgc gtagtgagat
     1621 ctatatctat gatctcgcag tctccggcga gcaccggagg cagggcattg ccaccgcgct
     1681 catcaatctc ctcaagcatg aggccaacgc gcttggtgct tatgtgatct acgtgcaagc
     1741 agattacggt gacgatcccg cagtggctct ctatacaaag ttgggcatac gggaagaagt
     1801 gatgcacttt gatatcgacc caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg
     1861 cttacgcgtc caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1921 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1981 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2041 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2101 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2161 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2221 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2281 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2341 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2401 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2461 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2521 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2581 ctcacatgtt ctttcctgca ctagatcccc ggcaagttca tccttagggt tttcatatct
     2641 cttaaaaagt caatttctgc ttgattaatt tcttgtggat tattgttttt gcggtttttc
     2701 attttttatc tccttgagta ttgaattttt tcttatatat aaaatatagc aaaaactatt
     2761 tttgccaact tttttacaaa aaatttttta ttaaaacact tagggctttg ctaaattctc
     2821 ttttaaaaga acttagtaaa gccctatgga tttaagaact gattatactt acgtagtaaa
     2881 aatactacag atattaatag tatagaccat atcaaaatca taatcaattt atatttatct
     2941 aaaattgtgg tccatacaaa atacaaaaaa attaggtttt tctaaattct attttaaaag
     3001 aacttagtaa cacctaatct ctaaattact tacaaaattc taaaattaga tttttagttc
     3061 ttcatatttc tttaaaagtt ttttaagaaa atctttttga ttttcaaaaa tttcatgcat
     3121 cataaaacta gtaaatttgg attttttttt gtaaaaatca taactttctt gggttttaag
     3181 ttgaaatctt aatggtttta ttggattttg cctagatttt tttaacgccg gactttcttt
     3241 atcttttata aaattataag aatttttaat accattttct attaaataat cttcttttat
     3301 aagcccctct tgcagtgcag ttgctaatct taaatactta tacgcttgag tttttgctac
     3361 atcaaagtct tttataaaat caagaaaagt tttataacca tcaaatttat aatattgatt
     3421 atcttttatt tcttttaaaa ttttaatatt ttccaccctg tggaatattt cttttttgaa
     3481 attgtcattt aacttttcct ttagtgtctt gtaataagaa attttacttt cattaatcat
     3541 taaattttgt tcaacataag tatctatctt ttttgttaaa actttaacaa cacgtttttt
     3601 ttccataaaa attttccttt atttttgcaa cattttcctt aattgatata attgttacta
     3661 ttttagattt tttaatatcc acttaagaat taatccctca ctagaatatt ttttattata
     3721 gaattcatat acatgttttt ctaaatctat aaatttatca acaattgctt tattataggg
     3781 tgtatctatc ttttcttttc ttaataatcg tctcatattc tttaaataac aacaaacact
     3841 gccggtttta aacctaaatt ctatgtaata tatttttgaa aaagtatata cattttcaac
     3901 cccgtttatt tcatacttta caaaaacatt atttattggt tttctataac catagtaaat
     3961 gcctaaaaat ttgtccccgt ctttcatagg atatagataa agtttttgaa tctctgattt
     4021 atctaataat ttttcaaaaa aaattataca ccgcttttgt ttgttactaa ctactctatg
     4081 caatgccttt aatatttttg tgtgatatat atttttatca cctttttttt ctttcagcag
     4141 gaagatactt tcttttggaa atttaatttt ttgttcattt attcttgcag ccttaagtcc
     4201 gattttcata tttaatcctt atatagcctt tctatttaat tcttttaaac tcaataaact
     4261 tttttggttt ttcattagtt ctaataattc gcaataatat gtgcaaaaca ctttattgta
     4321 ttctattttc ttttggttat tcaaatattc ttttataatt ggctttagaa gttcaatatt
     4381 tacttctttt tttagttgtt caataaggat attgaaaatg tttgttttta aatcttgata
     4441 attttgttga gaattttctt tttttctttc aattgacttc tctaatttac gctttatatt
     4501 gtttaaatcg ctatatttat gactttcaat aataaaatga ggcttaaatt tgtaattttc
     4561 gtatattttt tggaaacttg tttctaattg tttagaatta tattcatttt tttcaaattc
     4621 tttttgagtg ttgcatagaa tttcctttaa tttttcttgt ttttctttca tacaagattt
     4681 agtaaaatgt atatttttat tttttagtaa tttaatttct attattttta tgattttaag
     4741 tattttgatt atattatctt tatcaatatt taaattgaat aaaattgtaa gaaaaattga
     4801 aagaattttt ttacataaaa agttacattt attgaagtaa tttcttactt gatacttttc
     4861 tatttcatta atctttcttt cttcttttat attattttta ttatttaaac actcctttaa
     4921 atctacactc ccatttttag taaatctatc tttaaggccc cttgaaactc tattttgaaa
     4981 tcttaaatct tttttttctt gaaagtattt atagattttt tggtaacatt ctttttttga
     5041 aaaattaagt tgatagtata tttcggtccc caaatttacc cctaagtgtt tgtgataatt
     5101 agttgtgact ttaaattctt tttctaattt gtaaaggtaa ttttgcagtg tttttagttt
     5161 aacgggtgtt tgactatttc ttttaaggtt ttcattaaag taatagagta tgtttttttg
     5221 tgtgtatttc ttatatgttt tatttatgta cgctagtgtt gagataagga ctactaattt
     5281 atactgtaat ttattgtagc aagttgtggt tttactggtt ttgtttatat tagataagtt
     5341 ttccatttat agctccttga ataccggatt ttttatctat aaagtatata tatcacaatt
     5401 ttgatttaaa agtaaattct ttttttaaaa atacaaaaat tttttataaa ttatatagac
     5461 catgatattg tattaatact aattgatttt aatttaaggt tgaactaaaa taaacatagt
     5521 tttgtagaaa ataatttttc attattttgc tacttgatat tggatcgtaa aaatattagg
     5581 gctttacaaa gttcttttaa aagagaattt agtaaagccc taagtctttt aataaaaaat
     5641 tttttgtaaa aaagttggca aaaatagttt ttgctatata tttatatata agaaaaaatt
     5701 caatactcaa ggagataaaa aatggaaaac ttaaacaata atccacaaaa aattaatcaa
     5761 ataaataagc aaactaattt tattggtttt gaaaatttta tagaacttga taatcaagtt
     5821 aatcaaacta aagaaaaaag taaattaagt aagataggta agaaattacc cggaataagc
     5881 agtcaagtct agtgcggccg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5941 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6001 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6061 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca
     6121 tgattacgaa ttcgagctcg ttgatattaa acttaaaagc aatattattg ttgtaagaaa
     6181 tacgtaaacc ttaggctcta ttcctaggat tg
//
