LOCUS       sequence-318185-        6210 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 6210)
  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 6210)
  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_PresTL-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     25..41
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     25..41
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        760..835
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(853..870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(853..869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(862..884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1436..1969
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2072..2660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2561..2580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6137..6158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6173..6178,6179..6196,6197..6203)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattcgagct
       61 ccgaagttta ttattttatg atttaaatta ataaaaagat ttatcggtgt gaatagaata
      121 tttctttggt ttattatgca tcctagtaca tattatataa tttaatttaa tatataagca
      181 tattttttta aaatcagaaa ttttttactt agctaaagat tttaaacttg gtataattga
      241 attggaagag ccaaagtcaa cacttataga caaatatgat ctttctaacc ttcacgaaat
      301 agcatcaggg ggagcacctt tgtctaagga agtaggagag gcagttgcaa agaggtttca
      361 tttacccgga attaggcaag gatatgggtt aacagaaacc acatctgcaa ttttaattac
      421 acctgaagga gatgataaac caggagctgt tgggaaggtt gttccttttt ttgaggctaa
      481 agttgttgat ttagatactg gaaagacttt aggagttaat caaagaggcg agttatgtgt
      541 aagaggacct atgataatgt caggatacgt aaataatcca gaagctacaa atgcattgat
      601 agataaagat ggttggttac actcaggcga tatagcttat tgggacgaag atgaacactt
      661 ttttatagtt gatagattga aatctttgat taagtataaa ggttatcaag tagctccagc
      721 agaattggaa agtattttat tgcaacaccc agctcttccg ttttagagct agaaatagca
      781 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      841 ttgaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      901 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      961 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcggccgccc
     1021 taggttaata cccgagcttc aaggaagatt tcctattaag gttgaactta agagcttaag
     1081 catagacgat ttgaaaaaaa ttttaaaaca aacaaaaaat tctttaataa aacaatatgt
     1141 tgcgatgttt aaggtttatg atttagattt aaagtttagt gaggaagcta tagatagaat
     1201 tgcagagctt acttttaata tgaatcttga gagtgaaaat cttggtgcca gaagacttca
     1261 cggtgttatg gaaatagtgc ttgcagatct tttttttgaa gtgcctggca gtaagttgaa
     1321 aaaatttgaa ataaacttgg actatgttaa taaaaaaata caaattaacg aacaaaaaga
     1381 tttgaattat tatataattt agttaaaagc aattttaaat gagggaggtt tccatatgtt
     1441 acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc
     1501 aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc
     1561 tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc
     1621 ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt
     1681 cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc
     1741 gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga ggcagggcat
     1801 tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat
     1861 ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat
     1921 acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca
     1981 agccgagatc ggcttacgcg tccaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2701 gctggccttt tgctcacatg ttctttcctg cactagatcc ccggcaagtt catccttagg
     2761 gttttcatat ctcttaaaaa gtcaatttct gcttgattaa tttcttgtgg attattgttt
     2821 ttgcggtttt tcatttttta tctccttgag tattgaattt tttcttatat ataaaatata
     2881 gcaaaaacta tttttgccaa cttttttaca aaaaattttt tattaaaaca cttagggctt
     2941 tgctaaattc tcttttaaaa gaacttagta aagccctatg gatttaagaa ctgattatac
     3001 ttacgtagta aaaatactac agatattaat agtatagacc atatcaaaat cataatcaat
     3061 ttatatttat ctaaaattgt ggtccataca aaatacaaaa aaattaggtt tttctaaatt
     3121 ctattttaaa agaacttagt aacacctaat ctctaaatta cttacaaaat tctaaaatta
     3181 gatttttagt tcttcatatt tctttaaaag ttttttaaga aaatcttttt gattttcaaa
     3241 aatttcatgc atcataaaac tagtaaattt ggattttttt ttgtaaaaat cataactttc
     3301 ttgggtttta agttgaaatc ttaatggttt tattggattt tgcctagatt tttttaacgc
     3361 cggactttct ttatctttta taaaattata agaattttta ataccatttt ctattaaata
     3421 atcttctttt ataagcccct cttgcagtgc agttgctaat cttaaatact tatacgcttg
     3481 agtttttgct acatcaaagt cttttataaa atcaagaaaa gttttataac catcaaattt
     3541 ataatattga ttatctttta tttcttttaa aattttaata ttttccaccc tgtggaatat
     3601 ttcttttttg aaattgtcat ttaacttttc ctttagtgtc ttgtaataag aaattttact
     3661 ttcattaatc attaaatttt gttcaacata agtatctatc ttttttgtta aaactttaac
     3721 aacacgtttt ttttccataa aaattttcct ttatttttgc aacattttcc ttaattgata
     3781 taattgttac tattttagat tttttaatat ccacttaaga attaatccct cactagaata
     3841 ttttttatta tagaattcat atacatgttt ttctaaatct ataaatttat caacaattgc
     3901 tttattatag ggtgtatcta tcttttcttt tcttaataat cgtctcatat tctttaaata
     3961 acaacaaaca ctgccggttt taaacctaaa ttctatgtaa tatatttttg aaaaagtata
     4021 tacattttca accccgttta tttcatactt tacaaaaaca ttatttattg gttttctata
     4081 accatagtaa atgcctaaaa atttgtcccc gtctttcata ggatatagat aaagtttttg
     4141 aatctctgat ttatctaata atttttcaaa aaaaattata caccgctttt gtttgttact
     4201 aactactcta tgcaatgcct ttaatatttt tgtgtgatat atatttttat cacctttttt
     4261 ttctttcagc aggaagatac tttcttttgg aaatttaatt ttttgttcat ttattcttgc
     4321 agccttaagt ccgattttca tatttaatcc ttatatagcc tttctattta attcttttaa
     4381 actcaataaa cttttttggt ttttcattag ttctaataat tcgcaataat atgtgcaaaa
     4441 cactttattg tattctattt tcttttggtt attcaaatat tcttttataa ttggctttag
     4501 aagttcaata tttacttctt tttttagttg ttcaataagg atattgaaaa tgtttgtttt
     4561 taaatcttga taattttgtt gagaattttc tttttttctt tcaattgact tctctaattt
     4621 acgctttata ttgtttaaat cgctatattt atgactttca ataataaaat gaggcttaaa
     4681 tttgtaattt tcgtatattt tttggaaact tgtttctaat tgtttagaat tatattcatt
     4741 tttttcaaat tctttttgag tgttgcatag aatttccttt aatttttctt gtttttcttt
     4801 catacaagat ttagtaaaat gtatattttt attttttagt aatttaattt ctattatttt
     4861 tatgatttta agtattttga ttatattatc tttatcaata tttaaattga ataaaattgt
     4921 aagaaaaatt gaaagaattt ttttacataa aaagttacat ttattgaagt aatttcttac
     4981 ttgatacttt tctatttcat taatctttct ttcttctttt atattatttt tattatttaa
     5041 acactccttt aaatctacac tcccattttt agtaaatcta tctttaaggc cccttgaaac
     5101 tctattttga aatcttaaat cttttttttc ttgaaagtat ttatagattt tttggtaaca
     5161 ttcttttttt gaaaaattaa gttgatagta tatttcggtc cccaaattta cccctaagtg
     5221 tttgtgataa ttagttgtga ctttaaattc tttttctaat ttgtaaaggt aattttgcag
     5281 tgtttttagt ttaacgggtg tttgactatt tcttttaagg ttttcattaa agtaatagag
     5341 tatgtttttt tgtgtgtatt tcttatatgt tttatttatg tacgctagtg ttgagataag
     5401 gactactaat ttatactgta atttattgta gcaagttgtg gttttactgg ttttgtttat
     5461 attagataag ttttccattt atagctcctt gaataccgga ttttttatct ataaagtata
     5521 tatatcacaa ttttgattta aaagtaaatt ctttttttaa aaatacaaaa attttttata
     5581 aattatatag accatgatat tgtattaata ctaattgatt ttaatttaag gttgaactaa
     5641 aataaacata gttttgtaga aaataatttt tcattatttt gctacttgat attggatcgt
     5701 aaaaatatta gggctttaca aagttctttt aaaagagaat ttagtaaagc cctaagtctt
     5761 ttaataaaaa attttttgta aaaaagttgg caaaaatagt ttttgctata tatttatata
     5821 taagaaaaaa ttcaatactc aaggagataa aaaatggaaa acttaaacaa taatccacaa
     5881 aaaattaatc aaataaataa gcaaactaat tttattggtt ttgaaaattt tatagaactt
     5941 gataatcaag ttaatcaaac taaagaaaaa agtaaattaa gtaagatagg taagaaatta
     6001 cccggaataa gcagtcaagt ctagtgcggc cgcgcccaat acgcaaaccg cctctccccg
     6061 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6121 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     6181 ttatgcttcc ggctcgtatg ttgtgtggaa
//
