LOCUS       sequence-311496-        6148 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 6148)
  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 6148)
  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_P0026-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6148
                     /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        698..773
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(791..808)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(791..807)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(800..822)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1374..1907
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2010..2598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2499..2518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6075..6096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6111..6116,6117..6134,6135..6141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattcgagct
       61 ctattctcca ttcttttaaa attattatcc ttgtttatta tagtttaaaa taaatttaat
      121 tacaatttga aatttggtcg tttgactttc cttatattga caatctaagt ataatattaa
      181 gggaagagcc aaagtcaaca cttatagaca aatatgatct ttctaacctt cacgaaatag
      241 catcaggggg agcacctttg tctaaggaag taggagaggc agttgcaaag aggtttcatt
      301 tacccggaat taggcaagga tatgggttaa cagaaaccac atctgcaatt ttaattacac
      361 ctgaaggaga tgataaacca ggagctgttg ggaaggttgt tccttttttt gaggctaaag
      421 ttgttgattt agatactgga aagactttag gagttaatca aagaggcgag ttatgtgtaa
      481 gaggacctat gataatgtca ggatacgtaa ataatccaga agctacaaat gcattgatag
      541 ataaagatgg ttggttacac tcaggcgata tagcttattg ggacgaagat gaacactttt
      601 ttatagttga tagattgaaa tctttgatta agtataaagg ttatcaagta gctccagcag
      661 aattggaaag tattttattg caacacccag ctcttccgtt ttagagctag aaatagcaag
      721 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
      781 gaagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      841 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      901 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ggccgcccta
      961 ggttaatacc cgagcttcaa ggaagatttc ctattaaggt tgaacttaag agcttaagca
     1021 tagacgattt gaaaaaaatt ttaaaacaaa caaaaaattc tttaataaaa caatatgttg
     1081 cgatgtttaa ggtttatgat ttagatttaa agtttagtga ggaagctata gatagaattg
     1141 cagagcttac ttttaatatg aatcttgaga gtgaaaatct tggtgccaga agacttcacg
     1201 gtgttatgga aatagtgctt gcagatcttt tttttgaagt gcctggcagt aagttgaaaa
     1261 aatttgaaat aaacttggac tatgttaata aaaaaataca aattaacgaa caaaaagatt
     1321 tgaattatta tataatttag ttaaaagcaa ttttaaatga gggaggtttc catatgttac
     1381 gcagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa
     1441 gtatgggcat cattcgcaca tgtaggctcg gccctgacca agtcaaatcc atgcgggctg
     1501 ctcttgatct tttcggtcgt gagttcggag acgtagccac ctactcccaa catcagccgg
     1561 actccgatta cctcgggaac ttgctccgta gtaagacatt catcgcgctt gctgccttcg
     1621 accaagaagc ggttgttggc gctctcgcgg cttacgttct gcccaagttt gagcagccgc
     1681 gtagtgagat ctatatctat gatctcgcag tctccggcga gcaccggagg cagggcattg
     1741 ccaccgcgct catcaatctc ctcaagcatg aggccaacgc gcttggtgct tatgtgatct
     1801 acgtgcaagc agattacggt gacgatcccg cagtggctct ctatacaaag ttgggcatac
     1861 gggaagaagt gatgcacttt gatatcgacc caagtaccgc cacctaacaa ttcgttcaag
     1921 ccgagatcgg cttacgcgtc caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2641 tggccttttg ctcacatgtt ctttcctgca ctagatcccc ggcaagttca tccttagggt
     2701 tttcatatct cttaaaaagt caatttctgc ttgattaatt tcttgtggat tattgttttt
     2761 gcggtttttc attttttatc tccttgagta ttgaattttt tcttatatat aaaatatagc
     2821 aaaaactatt tttgccaact tttttacaaa aaatttttta ttaaaacact tagggctttg
     2881 ctaaattctc ttttaaaaga acttagtaaa gccctatgga tttaagaact gattatactt
     2941 acgtagtaaa aatactacag atattaatag tatagaccat atcaaaatca taatcaattt
     3001 atatttatct aaaattgtgg tccatacaaa atacaaaaaa attaggtttt tctaaattct
     3061 attttaaaag aacttagtaa cacctaatct ctaaattact tacaaaattc taaaattaga
     3121 tttttagttc ttcatatttc tttaaaagtt ttttaagaaa atctttttga ttttcaaaaa
     3181 tttcatgcat cataaaacta gtaaatttgg attttttttt gtaaaaatca taactttctt
     3241 gggttttaag ttgaaatctt aatggtttta ttggattttg cctagatttt tttaacgccg
     3301 gactttcttt atcttttata aaattataag aatttttaat accattttct attaaataat
     3361 cttcttttat aagcccctct tgcagtgcag ttgctaatct taaatactta tacgcttgag
     3421 tttttgctac atcaaagtct tttataaaat caagaaaagt tttataacca tcaaatttat
     3481 aatattgatt atcttttatt tcttttaaaa ttttaatatt ttccaccctg tggaatattt
     3541 cttttttgaa attgtcattt aacttttcct ttagtgtctt gtaataagaa attttacttt
     3601 cattaatcat taaattttgt tcaacataag tatctatctt ttttgttaaa actttaacaa
     3661 cacgtttttt ttccataaaa attttccttt atttttgcaa cattttcctt aattgatata
     3721 attgttacta ttttagattt tttaatatcc acttaagaat taatccctca ctagaatatt
     3781 ttttattata gaattcatat acatgttttt ctaaatctat aaatttatca acaattgctt
     3841 tattataggg tgtatctatc ttttcttttc ttaataatcg tctcatattc tttaaataac
     3901 aacaaacact gccggtttta aacctaaatt ctatgtaata tatttttgaa aaagtatata
     3961 cattttcaac cccgtttatt tcatacttta caaaaacatt atttattggt tttctataac
     4021 catagtaaat gcctaaaaat ttgtccccgt ctttcatagg atatagataa agtttttgaa
     4081 tctctgattt atctaataat ttttcaaaaa aaattataca ccgcttttgt ttgttactaa
     4141 ctactctatg caatgccttt aatatttttg tgtgatatat atttttatca cctttttttt
     4201 ctttcagcag gaagatactt tcttttggaa atttaatttt ttgttcattt attcttgcag
     4261 ccttaagtcc gattttcata tttaatcctt atatagcctt tctatttaat tcttttaaac
     4321 tcaataaact tttttggttt ttcattagtt ctaataattc gcaataatat gtgcaaaaca
     4381 ctttattgta ttctattttc ttttggttat tcaaatattc ttttataatt ggctttagaa
     4441 gttcaatatt tacttctttt tttagttgtt caataaggat attgaaaatg tttgttttta
     4501 aatcttgata attttgttga gaattttctt tttttctttc aattgacttc tctaatttac
     4561 gctttatatt gtttaaatcg ctatatttat gactttcaat aataaaatga ggcttaaatt
     4621 tgtaattttc gtatattttt tggaaacttg tttctaattg tttagaatta tattcatttt
     4681 tttcaaattc tttttgagtg ttgcatagaa tttcctttaa tttttcttgt ttttctttca
     4741 tacaagattt agtaaaatgt atatttttat tttttagtaa tttaatttct attattttta
     4801 tgattttaag tattttgatt atattatctt tatcaatatt taaattgaat aaaattgtaa
     4861 gaaaaattga aagaattttt ttacataaaa agttacattt attgaagtaa tttcttactt
     4921 gatacttttc tatttcatta atctttcttt cttcttttat attattttta ttatttaaac
     4981 actcctttaa atctacactc ccatttttag taaatctatc tttaaggccc cttgaaactc
     5041 tattttgaaa tcttaaatct tttttttctt gaaagtattt atagattttt tggtaacatt
     5101 ctttttttga aaaattaagt tgatagtata tttcggtccc caaatttacc cctaagtgtt
     5161 tgtgataatt agttgtgact ttaaattctt tttctaattt gtaaaggtaa ttttgcagtg
     5221 tttttagttt aacgggtgtt tgactatttc ttttaaggtt ttcattaaag taatagagta
     5281 tgtttttttg tgtgtatttc ttatatgttt tatttatgta cgctagtgtt gagataagga
     5341 ctactaattt atactgtaat ttattgtagc aagttgtggt tttactggtt ttgtttatat
     5401 tagataagtt ttccatttat agctccttga ataccggatt ttttatctat aaagtatata
     5461 tatcacaatt ttgatttaaa agtaaattct ttttttaaaa atacaaaaat tttttataaa
     5521 ttatatagac catgatattg tattaatact aattgatttt aatttaaggt tgaactaaaa
     5581 taaacatagt tttgtagaaa ataatttttc attattttgc tacttgatat tggatcgtaa
     5641 aaatattagg gctttacaaa gttcttttaa aagagaattt agtaaagccc taagtctttt
     5701 aataaaaaat tttttgtaaa aaagttggca aaaatagttt ttgctatata tttatatata
     5761 agaaaaaatt caatactcaa ggagataaaa aatggaaaac ttaaacaata atccacaaaa
     5821 aattaatcaa ataaataagc aaactaattt tattggtttt gaaaatttta tagaacttga
     5881 taatcaagtt aatcaaacta aagaaaaaag taaattaagt aagataggta agaaattacc
     5941 cggaataagc agtcaagtct agtgcggccg cgcccaatac gcaaaccgcc tctccccgcg
     6001 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6061 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6121 atgcttccgg ctcgtatgtt gtgtggaa
//
