LOCUS       sequence-311185-        6135 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 6135)
  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 6135)
  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_PresTS-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    348..369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(384..389,390..407,408..414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    422..438
                     /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     427..449
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     446..462
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        1106..1181
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1199..1216)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1199..1215)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1208..1230)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1782..2315
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2418..3006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2907..2926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaaaaatag tttttgctat atatttatat ataagaaaaa attcaatact caaggagata
       61 aaaaatggaa aacttaaaca ataatccaca aaaaattaat caaataaata agcaaactaa
      121 ttttattggt tttgaaaatt ttatagaact tgataatcaa gttaatcaaa ctaaagaaaa
      181 aagtaaatta agtaagatag gtaagaaatt acccggaata agcagtcaag tctagtgcgg
      241 ccgcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      301 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      361 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      421 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattcgagc
      481 tccgaagttt attattttat gatttaaatt aataaaaaga tttatcggtg tgaatagaat
      541 atttctttgg tttattatgc atcctagtac atattatata atttaatttg gaagagccaa
      601 agtcaacact tatagacaaa tatgatcttt ctaaccttca cgaaatagca tcagggggag
      661 cacctttgtc taaggaagta ggagaggcag ttgcaaagag gtttcattta cccggaatta
      721 ggcaaggata tgggttaaca gaaaccacat ctgcaatttt aattacacct gaaggagatg
      781 ataaaccagg agctgttggg aaggttgttc ctttttttga ggctaaagtt gttgatttag
      841 atactggaaa gactttagga gttaatcaaa gaggcgagtt atgtgtaaga ggacctatga
      901 taatgtcagg atacgtaaat aatccagaag ctacaaatgc attgatagat aaagatggtt
      961 ggttacactc aggcgatata gcttattggg acgaagatga acactttttt atagttgata
     1021 gattgaaatc tttgattaag tataaaggtt atcaagtagc tccagcagaa ttggaaagta
     1081 ttttattgca acacccagct cttccgtttt agagctagaa atagcaagtt aaaataaggc
     1141 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttga agcttggcac
     1201 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1261 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1321 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcgg ccgccctagg ttaatacccg
     1381 agcttcaagg aagatttcct attaaggttg aacttaagag cttaagcata gacgatttga
     1441 aaaaaatttt aaaacaaaca aaaaattctt taataaaaca atatgttgcg atgtttaagg
     1501 tttatgattt agatttaaag tttagtgagg aagctataga tagaattgca gagcttactt
     1561 ttaatatgaa tcttgagagt gaaaatcttg gtgccagaag acttcacggt gttatggaaa
     1621 tagtgcttgc agatcttttt tttgaagtgc ctggcagtaa gttgaaaaaa tttgaaataa
     1681 acttggacta tgttaataaa aaaatacaaa ttaacgaaca aaaagatttg aattattata
     1741 taatttagtt aaaagcaatt ttaaatgagg gaggtttcca tatgttacgc agcagcaacg
     1801 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
     1861 ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
     1921 tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc
     1981 tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg
     2041 ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct
     2101 atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca
     2161 tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag
     2221 attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga
     2281 tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct
     2341 tacgcgtcca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3061 cacatgttct ttcctgcact agatccccgg caagttcatc cttagggttt tcatatctct
     3121 taaaaagtca atttctgctt gattaatttc ttgtggatta ttgtttttgc ggtttttcat
     3181 tttttatctc cttgagtatt gaattttttc ttatatataa aatatagcaa aaactatttt
     3241 tgccaacttt tttacaaaaa attttttatt aaaacactta gggctttgct aaattctctt
     3301 ttaaaagaac ttagtaaagc cctatggatt taagaactga ttatacttac gtagtaaaaa
     3361 tactacagat attaatagta tagaccatat caaaatcata atcaatttat atttatctaa
     3421 aattgtggtc catacaaaat acaaaaaaat taggtttttc taaattctat tttaaaagaa
     3481 cttagtaaca cctaatctct aaattactta caaaattcta aaattagatt tttagttctt
     3541 catatttctt taaaagtttt ttaagaaaat ctttttgatt ttcaaaaatt tcatgcatca
     3601 taaaactagt aaatttggat ttttttttgt aaaaatcata actttcttgg gttttaagtt
     3661 gaaatcttaa tggttttatt ggattttgcc tagatttttt taacgccgga ctttctttat
     3721 cttttataaa attataagaa tttttaatac cattttctat taaataatct tcttttataa
     3781 gcccctcttg cagtgcagtt gctaatctta aatacttata cgcttgagtt tttgctacat
     3841 caaagtcttt tataaaatca agaaaagttt tataaccatc aaatttataa tattgattat
     3901 cttttatttc ttttaaaatt ttaatatttt ccaccctgtg gaatatttct tttttgaaat
     3961 tgtcatttaa cttttccttt agtgtcttgt aataagaaat tttactttca ttaatcatta
     4021 aattttgttc aacataagta tctatctttt ttgttaaaac tttaacaaca cgtttttttt
     4081 ccataaaaat tttcctttat ttttgcaaca ttttccttaa ttgatataat tgttactatt
     4141 ttagattttt taatatccac ttaagaatta atccctcact agaatatttt ttattataga
     4201 attcatatac atgtttttct aaatctataa atttatcaac aattgcttta ttatagggtg
     4261 tatctatctt ttcttttctt aataatcgtc tcatattctt taaataacaa caaacactgc
     4321 cggttttaaa cctaaattct atgtaatata tttttgaaaa agtatataca ttttcaaccc
     4381 cgtttatttc atactttaca aaaacattat ttattggttt tctataacca tagtaaatgc
     4441 ctaaaaattt gtccccgtct ttcataggat atagataaag tttttgaatc tctgatttat
     4501 ctaataattt ttcaaaaaaa attatacacc gcttttgttt gttactaact actctatgca
     4561 atgcctttaa tatttttgtg tgatatatat ttttatcacc ttttttttct ttcagcagga
     4621 agatactttc ttttggaaat ttaatttttt gttcatttat tcttgcagcc ttaagtccga
     4681 ttttcatatt taatccttat atagcctttc tatttaattc ttttaaactc aataaacttt
     4741 tttggttttt cattagttct aataattcgc aataatatgt gcaaaacact ttattgtatt
     4801 ctattttctt ttggttattc aaatattctt ttataattgg ctttagaagt tcaatattta
     4861 cttctttttt tagttgttca ataaggatat tgaaaatgtt tgtttttaaa tcttgataat
     4921 tttgttgaga attttctttt tttctttcaa ttgacttctc taatttacgc tttatattgt
     4981 ttaaatcgct atatttatga ctttcaataa taaaatgagg cttaaatttg taattttcgt
     5041 atattttttg gaaacttgtt tctaattgtt tagaattata ttcatttttt tcaaattctt
     5101 tttgagtgtt gcatagaatt tcctttaatt tttcttgttt ttctttcata caagatttag
     5161 taaaatgtat atttttattt tttagtaatt taatttctat tatttttatg attttaagta
     5221 ttttgannat attatcttta tcaatattta aattgaataa aattgtaaga aaaattgaaa
     5281 gaattttttt acataaaaag ttacatttat tgaagtaatt tcttacttga tacttttcta
     5341 tttcattaat ctttctttct tcttttatat tatttttatt atttaaacac tcctttaaat
     5401 ctacactccc atttttagta aatctatctt taaggcccct tgaaactcta ttttgaaatc
     5461 ttaaatcttt tttttcttga aagtatttat agattttttg gtaacattct ttttttgaaa
     5521 aattaagttg atagtatatt tcggtcccca aatttacccc taagtgtttg tgataattag
     5581 ttgtgacttt aaattctttt tctaatttgt aaaggtaatt ttgcagtgtt tttagtttaa
     5641 cgggtgtttg actatttctt ttaaggtttt cattaaagta atagagtatg tttttttgtg
     5701 tgtatttctt atatgtttta tttatgtacg ctagtgttga gataaggact actaatttat
     5761 actgtaattt attgtagcaa gttgtggttt tactggtttt gtttatatta gataagtttt
     5821 ccatttatag ctccttgaat accggatttt ttatctataa agtatatata tcacaatttt
     5881 gatttaaaag taaattcttt ttttaaaaat acaaaaattt tttataaatt atatagacca
     5941 tgatattgta ttaatactaa ttgattttaa tttaaggttg aactaaaata aacatagttt
     6001 tgtagaaaat aatttttcat tattttgcta cttgatattg gatcgtaaaa atattagggc
     6061 tttacaaagt tcttttaaaa gagaatttag taaagcccta agtcttttaa taaaaaattt
     6121 tttgtaaaaa agttg
//
