LOCUS       sequence-306830-        6593 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 6593)
  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 6593)
  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: pBSV2B_Psyn-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2811..2832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2847..2852,2853..2870,2871..2877)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2885..2901
                     /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     2890..2912
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2909..2925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2909..2925
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2961..2995
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        3512..3587
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3630..3647)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3630..3646)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3639..3661)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5339..5927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5828..5847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttttattgg attttgccta gattttttta acgccggact ttctttatct tttataaaat
       61 tataagaatt tttaatacca ttttctatta aataatcttc ttttataagc ccctcttgca
      121 gtgcagttgc taatcttaaa tacttatacg cttgagtttt tgctacatca aagtctttta
      181 taaaatcaag aaaagtttta taaccatcaa atttataata ttgattatct tttatttctt
      241 ttaaaatttt aatattttcc accctgtgga atatttcttt tttgaaattg tcatttaact
      301 tttcctttag tgtcttgtaa taagaaattt tactttcatt aatcattaaa ttttgttcaa
      361 cataagtatc tatctttttt gttaaaactt taacaacacg ttttttttcc ataaaaattt
      421 tcctttattt ttgcaacatt ttccttaatt gatataattg ttactatttt agatttttta
      481 atatccactt aagaattaat ccctcactag aatatttttt attatagaat tcatatacat
      541 gtttttctaa atctataaat ttatcaacaa ttgctttatt atagggtgta tctatctttt
      601 cttttcttaa taatcgtctc atattcttta aataacaaca aacactgccg gttttaaacc
      661 taaattctat gtaatatatt tttgaaaaag tatatacatt ttcaaccccg tttatttcat
      721 actttacaaa aacattattt attggttttc tataaccata gtaaatgcct aaaaatttgt
      781 ccccgtcttt cataggatat agataaagtt tttgaatctc tgatttatct aataattttt
      841 caaaaaaaat tatacaccgc ttttgtttgt tactaactac tctatgcaat gcctttaata
      901 tttttgtgtg atatatattt ttatcacctt ttttttcttt cagcaggaag atactttctt
      961 ttggaaattt aattttttgt tcatttattc ttgcagcctt aagtccgatt ttcatattta
     1021 atccttatat agcctttcta tttaattctt ttaaactcaa taaacttttt tggtttttca
     1081 ttagttctaa taattcgcaa taatatgtgc aaaacacttt attgtattct attttctttt
     1141 ggttattcaa atattctttt ataattggct ttagaagttc aatatttact tcttttttta
     1201 gttgttcaat aaggatattg aaaatgtttg tttttaaatc ttgataattt tgttgagaat
     1261 tttctttttt tctttcaatt gacttctcta atttacgctt tatattgttt aaatcgctat
     1321 atttatgact ttcaataata aaatgaggct taaatttgta attttcgtat attttttgga
     1381 aacttgtttc taattgttta gaattatatt catttttttc aaattctttt tgagtgttgc
     1441 atagaatttc ctttaatttt tcttgttttt ctttcataca agatttagta aaatgtatat
     1501 ttttattttt tagtaattta atttctatta tttttatgat tttaagtatt ttgattatat
     1561 tatctttatc aatatttaaa ttgaataaaa ttgtaagaaa aattgaaaga atttttttac
     1621 ataaaaagtt acatttattg aagtaatttc ttacttgata cttttctatt tcattaatct
     1681 ttctttcttc ttttatatta tttttattat ttaaacactc ctttaaatct acactcccat
     1741 ttttagtaaa tctatcttta aggccccttg aaactctatt ttgaaatctt aaatcttttt
     1801 tttcttgaaa gtatttatag attttttggt aacattcttt ttttgaaaaa ttaagttgat
     1861 agtatatttc ggtccccaaa tttaccccta agtgtttgtg ataattagtt gtgactttaa
     1921 attctttttc taatttgtaa aggtaatttt gcagtgtttt tagtttaacg ggtgtttgac
     1981 tatttctttt aaggttttca ttaaagtaat agagtatgtt tttttgtgtg tatttcttat
     2041 atgttttatt tatgtacgct agtgttgaga taaggactac taatttatac tgtaatttat
     2101 tgtagcaagt tgtggtttta ctggttttgt ttatattaga taagttttcc atttatagct
     2161 ccttgaatac cggatttttt atctataaag tatatatatc acaattttga tttaaaagta
     2221 aattcttttt ttaaaaatac aaaaattttt tataaattat atagaccatg atattgtatt
     2281 aatactaatt gattttaatt taaggttgaa ctaaaataaa catagttttg tagaaaataa
     2341 tttttcatta ttttgctact tgatattgga tcgtaaaaat attagggctt tacaaagttc
     2401 ttttaaaaga gaatttagta aagccctaag tcttttaata aaaaattttt tgtaaaaaag
     2461 ttggcaaaaa tagtttttgc tatatattta tatataagaa aaaattcaat actcaaggag
     2521 ataaaaaatg gaaaacttaa acaataatcc acaaaaaatt aatcaaataa ataagcaaac
     2581 taattttatt ggttttgaaa attttataga acttgataat caagttaatc aaactaaaga
     2641 aaaaagtaaa ttaagtaaga taggtaagaa attacccgga ataagcagtc aagtctagtg
     2701 cggccgcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2761 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     2821 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2881 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattcg
     2941 agctcgaatt ctaaagatct ttgacagcta gctcagtcct aggtataata ctagtggaag
     3001 agccaaagtc aacacttata gacaaatatg atctttctaa ccttcacgaa atagcatcag
     3061 ggggagcacc tttgtctaag gaagtaggag aggcagttgc aaagaggttt catttacccg
     3121 gaattaggca aggatatggg ttaacagaaa ccacatctgc aattttaatt acacctgaag
     3181 gagatgataa accaggagct gttgggaagg ttgttccttt ttttgaggct aaagttgttg
     3241 atttagatac tggaaagact ttaggagtta atcaaagagg cgagttatgt gtaagaggac
     3301 ctatgataat gtcaggatac gtaaataatc cagaagctac aaatgcattg atagataaag
     3361 atggttggtt acactcaggc gatatagctt attgggacga agatgaacac ttttttatag
     3421 ttgatagatt gaaatctttg attaagtata aaggttatca agtagctcca gcagaattgg
     3481 aaagtatttt attgcaacac ccagctcttc cgttttagag ctagaaatag caagttaaaa
     3541 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgaagct
     3601 tgggcccgaa caaaaactca aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     3661 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3721 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3781 gaatggcgcg gccgccctag gttaatcttc aataacatgt aaaccacgac gttttaaatc
     3841 ttctaatgaa tctaacattc ctcttattaa ttcaacagga tgtccttccc aatcttcaac
     3901 aacaccaaca attcttaaag gttcacaagt tctataagat tgtgttggat taccaggaaa
     3961 ttttttattt gtaagatttg gatcatcttc aaatggtcct gttggttcaa ctatatatat
     4021 ataaccacga ccttcaagac cagatagtga catagcaagt tcagctcccc aaactgctgg
     4081 ttccattaaa gctgaaaaat aaatatgttt aagaattcga ccatcttcaa aatgagaaat
     4141 aaatcctgta gttagtaaat caccaattgc taaattagct ttagttccat gataaaatgg
     4201 tccttgtact tgtttataat tatcatgaga aataggaatc caatctttta ccatgacgtt
     4261 acctcctcat gatgtttaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     4321 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     4381 gcatagactg taccccaaaa aaacagtcat aacaagccat gcggccgtac ccgagcttca
     4441 aggaagattt cctattaagg ttgaacttaa gagcttaagc atagacgatt tgaaaaaaat
     4501 tttaaaacaa acaaaaaatt ctttaataaa acaatatgtt gcgatgttta aggtttatga
     4561 tttagattta aagtttagtg aggaagctat agatagaatt gcagagctta cttttaatat
     4621 gaatcttgag agtgaaaatc ttggtgccag aagacttcac ggtgttatgg aaatagtgct
     4681 tgcagatctt ttttttgaag tgcctggcag taagttgaaa aaatttgaaa taaacttgga
     4741 ctatgttaat aaaaaaatac aaattaacga acaaaaagat ttgaattatt atataattta
     4801 gttaaaagca attttaaatg agggaggttt ccatcatatg gctaaacctt taagtcaaga
     4861 agaaagtact ttaattgaaa gagctactgc tactattaat agtattccta ttagtgaaga
     4921 ttatagtgtt gctagtgctg ctttaagtag tgatggtaga atttttactg gtgttaatgt
     4981 ttatcatttt actggtggtc cttgtgctga attagttgtt ttaggtactg ccgccgctgc
     5041 tgctgctggt aatctcactt gtattgttgc tattggtaat gaaaatagag gtattttaag
     5101 tccttgtggt agatgtagac aagttttatt agatttacat cctggtatta aagctattgt
     5161 taaagatagt gatggtcaac ctactgctgt tggtattaga gaattattac ctagtggtta
     5221 tgtttgggaa ggttaacaat tcgttcaagc cgagatcggc ttacgcgtcc aaaatccctt
     5281 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5341 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5401 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5461 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5521 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5581 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5641 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5701 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5761 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5821 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5881 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5941 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcac
     6001 tagatccccg gcaagttcat ccttagggtt ttcatatctc ttaaaaagtc aatttctgct
     6061 tgattaattt cttgtggatt attgtttttg cggtttttca ttttttatct ccttgagtat
     6121 tgaatttttt cttatatata aaatatagca aaaactattt ttgccaactt ttttacaaaa
     6181 aattttttat taaaacactt agggctttgc taaattctct tttaaaagaa cttagtaaag
     6241 ccctatggat ttaagaactg attatactta cgtagtaaaa atactacaga tattaatagt
     6301 atagaccata tcaaaatcat aatcaattta tatttatcta aaattgtggt ccatacaaaa
     6361 tacaaaaaaa ttaggttttt ctaaattcta ttttaaaaga acttagtaac acctaatctc
     6421 taaattactt acaaaattct aaaattagat ttttagttct tcatatttct ttaaaagttt
     6481 tttaagaaaa tctttttgat tttcaaaaat ttcatgcatc ataaaactag taaatttgga
     6541 tttttttttg taaaaatcat aactttcttg ggttttaagt tgaaatctta atg
//
