LOCUS       sequence-311480-        6240 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 6240)
  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 6240)
  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_P0826L-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..76
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(94..111)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(94..110)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(103..125)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             677..1210
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1313..1901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1802..1821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5378..5399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5414..5419,5420..5437,5438..5444)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5452..5468
                     /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     5457..5479
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5476..5492
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5476..5492
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
       61 ggcaccgagt cggtgctttt tttgaagctt ggcactggcc gtcgttttac aacgtcgtga
      121 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      181 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      241 tggcgaatgg cgcggccgcc ctaggttaat acccgagctt caaggaagat ttcctattaa
      301 ggttgaactt aagagcttaa gcatagacga tttgaaaaaa attttaaaac aaacaaaaaa
      361 ttctttaata aaacaatatg ttgcgatgtt taaggtttat gatttagatt taaagtttag
      421 tgaggaagct atagatagaa ttgcagagct tacttttaat atgaatcttg agagtgaaaa
      481 tcttggtgcc agaagacttc acggtgttat ggaaatagtg cttgcagatc ttttttttga
      541 agtgcctggc agtaagttga aaaaatttga aataaacttg gactatgtta ataaaaaaat
      601 acaaattaac gaacaaaaag atttgaatta ttatataatt tagttaaaag caattttaaa
      661 tgagggaggt ttccatatgt tacgcagcag caacgatgtt acgcagcagg gcagtcgccc
      721 taaaacaaag ttaggtggct caagtatggg catcattcgc acatgtaggc tcggccctga
      781 ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt cgtgagttcg gagacgtagc
      841 cacctactcc caacatcagc cggactccga ttacctcggg aacttgctcc gtagtaagac
      901 attcatcgcg cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt
      961 tctgcccaag tttgagcagc cgcgtagtga gatctatatc tatgatctcg cagtctccgg
     1021 cgagcaccgg aggcagggca ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa
     1081 cgcgcttggt gcttatgtga tctacgtgca agcagattac ggtgacgatc ccgcagtggc
     1141 tctctataca aagttgggca tacgggaaga agtgatgcac tttgatatcg acccaagtac
     1201 cgccacctaa caattcgttc aagccgagat cggcttacgc gtccaaaatc ccttaacgtg
     1261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1441 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcactagatc
     1981 cccggcaagt tcatccttag ggttttcata tctcttaaaa agtcaatttc tgcttgatta
     2041 atttcttgtg gattattgtt tttgcggttt ttcatttttt atctccttga gtattgaatt
     2101 ttttcttata tataaaatat agcaaaaact atttttgcca acttttttac aaaaaatttt
     2161 ttattaaaac acttagggct ttgctaaatt ctcttttaaa agaacttagt aaagccctat
     2221 ggatttaaga actgattata cttacgtagt aaaaatacta cagatattaa tagtatagac
     2281 catatcaaaa tcataatcaa tttatattta tctaaaattg tggtccatac aaaatacaaa
     2341 aaaattaggt ttttctaaat tctattttaa aagaacttag taacacctaa tctctaaatt
     2401 acttacaaaa ttctaaaatt agatttttag ttcttcatat ttctttaaaa gttttttaag
     2461 aaaatctttt tgattttcaa aaatttcatg catcataaaa ctagtaaatt tggatttttt
     2521 tttgtaaaaa tcataacttt cttgggtttt aagttgaaat cttaatggtt ttattggatt
     2581 ttgcctagat ttttttaacg ccggactttc tttatctttt ataaaattat aagaattttt
     2641 aataccattt tctattaaat aatcttcttt tataagcccc tcttgcagtg cagttgctaa
     2701 tcttaaatac ttatacgctt gagtttttgc tacatcaaag tcttttataa aatcaagaaa
     2761 agttttataa ccatcaaatt tataatattg attatctttt atttctttta aaattttaat
     2821 attttccacc ctgtggaata tttctttttt gaaattgtca tttaactttt cctttagtgt
     2881 cttgtaataa gaaattttac tttcattaat cattaaattt tgttcaacat aagtatctat
     2941 cttttttgtt aaaactttaa caacacgttt tttttccata aaaattttcc tttatttttg
     3001 caacattttc cttaattgat ataattgtta ctattttaga ttttttaata tccacttaag
     3061 aattaatccc tcactagaat attttttatt atagaattca tatacatgtt tttctaaatc
     3121 tataaattta tcaacaattg ctttattata gggtgtatct atcttttctt ttcttaataa
     3181 tcgtctcata ttctttaaat aacaacaaac actgccggtt ttaaacctaa attctatgta
     3241 atatattttt gaaaaagtat atacattttc aaccccgttt atttcatact ttacaaaaac
     3301 attatttatt ggttttctat aaccatagta aatgcctaaa aatttgtccc cgtctttcat
     3361 aggatataga taaagttttt gaatctctga tttatctaat aatttttcaa aaaaaattat
     3421 acaccgcttt tgtttgttac taactactct atgcaatgcc tttaatattt ttgtgtgata
     3481 tatattttta tcaccttttt tttctttcag caggaagata ctttcttttg gaaatttaat
     3541 tttttgttca tttattcttg cagccttaag tccgattttc atatttaatc cttatatagc
     3601 ctttctattt aattctttta aactcaataa acttttttgg tttttcatta gttctaataa
     3661 ttcgcaataa tatgtgcaaa acactttatt gtattctatt ttcttttggt tattcaaata
     3721 ttcttttata attggcttta gaagttcaat atttacttct ttttttagtt gttcaataag
     3781 gatattgaaa atgtttgttt ttaaatcttg ataattttgt tgagaatttt ctttttttct
     3841 ttcaattgac ttctctaatt tacgctttat attgtttaaa tcgctatatt tatgactttc
     3901 aataataaaa tgaggcttaa atttgtaatt ttcgtatatt ttttggaaac ttgtttctaa
     3961 ttgtttagaa ttatattcat ttttttcaaa ttctttttga gtgttgcata gaatttcctt
     4021 taatttttct tgtttttctt tcatacaaga tttagtaaaa tgtatatttt tattttttag
     4081 taatttaatt tctattattt ttatgatttt aagtattttg attatattat ctttatcaat
     4141 atttaaattg aataaaattg taagaaaaat tgaaagaatt tttttacata aaaagttaca
     4201 tttattgaag taatttctta cttgatactt ttctatttca ttaatctttc tttcttcttt
     4261 tatattattt ttattattta aacactcctt taaatctaca ctcccatttt tagtaaatct
     4321 atctttaagg ccccttgaaa ctctattttg aaatcttaaa tctttttttt cttgaaagta
     4381 tttatagatt ttttggtaac attctttttt tgaaaaatta agttgatagt atatttcggt
     4441 ccccaaattt acccctaagt gtttgtgata attagttgtg actttaaatt ctttttctaa
     4501 tttgtaaagg taattttgca gtgtttttag tttaacgggt gtttgactat ttcttttaag
     4561 gttttcatta aagtaataga gtatgttttt ttgtgtgtat ttcttatatg ttttatttat
     4621 gtacgctagt gttgagataa ggactactaa tttatactgt aatttattgt agcaagttgt
     4681 ggttttactg gttttgttta tattagataa gttttccatt tatagctcct tgaataccgg
     4741 attttttatc tataaagtat atatatcaca attttgattt aaaagtaaat tcttttttta
     4801 aaaatacaaa aattttttat aaattatata gaccatgata ttgtattaat actaattgat
     4861 tttaatttaa ggttgaacta aaataaacat agttttgtag aaaataattt ttcattattt
     4921 tgctacttga tattggatcg taaaaatatt agggctttac aaagttcttt taaaagagaa
     4981 tttagtaaag ccctaagtct tttaataaaa aattttttgt aaaaaagttg gcaaaaatag
     5041 tttttgctat atatttatat ataagaaaaa attcaatact caaggagata aaaaatggaa
     5101 aacttaaaca ataatccaca aaaaattaat caaataaata agcaaactaa ttttattggt
     5161 tttgaaaatt ttatagaact tgataatcaa gttaatcaaa ctaaagaaaa aagtaaatta
     5221 agtaagatag gtaagaaatt acccggaata agcagtcaag tctagtgcgg ccgcgcccaa
     5281 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5341 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     5401 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     5461 gataacaatt tcacacagga aacagctatg acatgattac gaattcgagc tcggcaatag
     5521 aagaatctat agaaagcata aaaagcctta tagaaaacaa agaatacata caaaagttac
     5581 attttataaa aaaattaata aataaggttt acaaaaaatt aaattacttt ttttaaaaac
     5641 taaactttga aagccttgtt ataatataaa atataataat caaatattat tcaaagttaa
     5701 cagcaatgaa gtttataata aattggaaga gccaaagtca acacttatag acaaatatga
     5761 tctttctaac cttcacgaaa tagcatcagg gggagcacct ttgtctaagg aagtaggaga
     5821 ggcagttgca aagaggtttc atttacccgg aattaggcaa ggatatgggt taacagaaac
     5881 cacatctgca attttaatta cacctgaagg agatgataaa ccaggagctg ttgggaaggt
     5941 tgttcctttt tttgaggcta aagttgttga tttagatact ggaaagactt taggagttaa
     6001 tcaaagaggc gagttatgtg taagaggacc tatgataatg tcaggatacg taaataatcc
     6061 agaagctaca aatgcattga tagataaaga tggttggtta cactcaggcg atatagctta
     6121 ttgggacgaa gatgaacact tttttatagt tgatagattg aaatctttga ttaagtataa
     6181 aggttatcaa gtagctccag cagaattgga aagtatttta ttgcaacacc cagctcttcc
//
