LOCUS       sequence-292420-        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"
     rep_origin      71..659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     560..579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4136..4157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4172..4177,4178..4195,4196..4202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4210..4226
                     /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     4215..4237
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4234..4250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4234..4250
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        4999..5074
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(5092..5109)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5092..5108)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5101..5123)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5675..6208
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
       61 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      181 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
      241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      661 acgccagcaa cgcggccttt ttacggttcc tgggcttttg ctggcctttt gctcacatgt
      721 tctttcctgc actagatccc cggcaagttc atccttaggg tttccatatc tcttaaaaag
      781 tcaatttctg cttgattaat ttcttgtgga ttattgtttt tgcggttttt cattttttat
      841 ctccttgagt attgaatttt ttcttatata taaatatata gcaaaaacta tttttgccaa
      901 cttttttaca aaaaattttt tattaaaaca cttagggctt tgctaaattc tcttttaaaa
      961 gaacttagta aagccctatg gatttaagaa ctgattatac ttacgtagta aaaatactac
     1021 agatattaat agtatagacc atatcaaaat cataatcaat ttatatttat ctaaaattgt
     1081 agtccataca aaatacaaaa aaattaggtt tttctaaatt ctattttaaa agaacttagt
     1141 aacacctaat ctctaaatta cttacaaaat tctaaaatta gatttttagt tcttcatatt
     1201 tctttaaaag ttttttaaga aaatcttttt gattttcaaa aatttcatgc atcataaaac
     1261 tagtaaattt ggattttttt ttgtaaaaat cataactttc ttgggtttta agttgaaatc
     1321 ttaatggttt tattggattt tgcctagatt tttttaacgc cggactttct ttatctttta
     1381 taaaattata agaattttta ataccatttt ctattaaata atcttctttt ataagcccct
     1441 cttgcagtgc agttgctaat cttaaatact tatacgcttg agtttttgct acatcaaagt
     1501 cttttataaa atcaagaaaa gttttataac catcaaattt ataatattga ttatctttta
     1561 tttcttttaa aattttaata ttttccaccc tgtggaatat ttcttttttg aaattgtcat
     1621 ttaacttttc ctttagtgtc ttgtaataag aaattttact ttcattaatc attaaatttt
     1681 gttcaacata agtatctatc ttttttgtta aaactttaac aacacgtttt ttttccataa
     1741 aaattttcct ttatttttgc aacattttcc ttaattgata taattgttac tattttagat
     1801 tttttaatat ccacttaaga attaatccct cactagaata ttttttatta tagaattcat
     1861 atacatgttt ttctaaatct ataaatttat caacaattgc tttattatag ggtgtatcta
     1921 tcttttcttt tcttaataat cgtctcatat tctttaaata acaacaaaca ctgccggttt
     1981 taaacctaaa ttctatgtaa tatatttttg aaaaagtata tacattttca accccgttta
     2041 tttcatactt tacaaaaaca ttatttattg gttttctata accatagtaa atgcctaaaa
     2101 atttgtcccc gtctttcata ggatatagat aaagtttttg aatctctgat ttatctaata
     2161 atttttcaaa aaaaattata caccgctttt gtttgttact aactactcta tgcaatgcct
     2221 ttaatatttt tgtgtgatat atatttttat cacctttttt ttctttcagc aggaagatac
     2281 tttcttttgg aaatttaatt ttttgttcat ttattcttgc agccttaagt ccgattttca
     2341 tatttaatcc ttatatagcc tttctattta attcttttaa actcaataaa cttttttggt
     2401 ttttcattag ttctaataat tcgcaataat atgtgcaaaa cactttattg tattctattt
     2461 tcttttggtt attcaaatat tcttttataa ttggctttag aagttcaata tttacttctt
     2521 tttttagttg ttcaataagg atattgaaaa tgtttgtttt taaatcttga taattttgtt
     2581 gagaattttc tttttttctt tcaattgact tctctaattt acgctttata ttgtttaaat
     2641 cgctatattt atgactttca ataataaaat gaggcttaaa tttgtaattt tcgtatattt
     2701 tttggaaact tgtttctaat tgtttagaat tatattcatt tttttcaaat tctttttgag
     2761 tgttgcatag aatttccttt aatttttctt gtttttcttt catacaagat ttagtaaaat
     2821 gtatattttt attttttagt aatttaattt ctattatttt tatgatttta agtattttga
     2881 ttatattatc tttatcaata tttaaattga ataaaattgt aagaaaaatt gaaagaattt
     2941 ttttacataa aaagttacat ttattgaagt aatttcttac ttgatacttt tctatttcat
     3001 taatctttct ttcttctttt atattatttt tattatttaa acactccttt aaatctacac
     3061 tcccattttt agtaaatcta tctttaaggc cccttgaaac tctattttga aatcttaaat
     3121 cttttttttc ttgaaagtat ttatagattt tttggtaaca ttcttttttt gaaaaattaa
     3181 gttgatagta tatttcggtc cccaaattta cccctaagtg tttgtgataa ttagttgtga
     3241 ctttaaattc tttttctaat ttgtaaaggt aattttgcag tgtttttagt ttaacgggtg
     3301 tttgactatt tcttttaagg ttttcattaa agtaatagag tatgtttttt tgtgtgtatt
     3361 tcttatatgt tttatttatg tacgctagtg ttgagataag aactactaat ttatactgta
     3421 atttattgta gcaagttgtg gttttactgg ttttgtttat attagataag ttttccattt
     3481 atagctcctt gaataccgga ttttttatct ataaagtata tatatcacaa ttttgattta
     3541 aaagtaaatt ctttttttaa aaatacaaaa attttttata aattatatag accatgatat
     3601 tgtattaata ctaattgatt ttaatttaag gttgaactaa aataaacata gttttgtaga
     3661 aaataatttt tcattatttt gctacttgat attggatcgt aaaaatatta gggctttaca
     3721 aagttctttt aaaagagaat tagtaaagcc ctaagtcttt taataaaaaa ttttttgtaa
     3781 aaaagttggc aaaaatagtt tttgctatat atttatatat aagaaaaaat tcaatactca
     3841 aggagataaa aaatggaaaa cttaaacaat aatccacaaa aaattaatca aataaataag
     3901 caaactaatt ttattggttt tgaaaatttt atagaacttg ataatcaagt taatcaaact
     3961 aaagaaaaaa gtaaattaag taagataggt aagaaattac ccggaataag cagtcaagtc
     4021 tagtgcggcc gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4081 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4141 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4201 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
     4261 attcgagctc ggcaatagaa gaatctatag aaagcataaa aagccttata gaaaacaaag
     4321 aatacataca aaagttacat tttataaaaa aattaataaa taaggtttac aaaaaattaa
     4381 attacttttt ttaaaaacta aactttgaaa gccttgttat aatataaaat ataataatca
     4441 aatattattc aaagttaaca gcaatgaagt ttataataaa ttggaagagc caaagtcaac
     4501 acttatagac aaatatgatc tttctaacct tcacgaaata gcatcagggg gagcaccttt
     4561 gtctaaggaa gtaggagagg cagttgcaaa gaggtttcat ttacccggaa ttaggcaagg
     4621 atatgggtta acagaaacca catctgcaat tttaattaca cctgaaggag atgataaacc
     4681 aggagctgtt gggaaggttg ttcctttttt tgaggctaaa gttgttgatt tagatactgg
     4741 aaagacttta ggagttaatc aaagaggcga gttatgtgta agaggaccta tgataatgtc
     4801 aggatacgta aataatccag aagctacaaa tgcattgata gataaagatg gttggttaca
     4861 ctcaggcgat atagcttatt gggacgaaga tgaacacttt tttatagttg atagattgaa
     4921 atctttgatt aagtataaag gttatcaagt agctccagca gaattggaaa gtattttatt
     4981 gcaacaccca gctcttccgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     5041 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgaagcttgg cactggccgt
     5101 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5161 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5221 acagttgcgc agcctgaatg gcgaatggcg cggccgccct aggttaatac ccgagcttca
     5281 aggaagattt cctattaagg ttgaacttaa gagcttaagc atagacgatt tgaaaaaaat
     5341 tttaaaacaa acaaaaaatt ctttaataaa acaatatgtt gcgatgttta aggtttatga
     5401 tttagattta aagtttagtg aggaagctat agatagaatt gcagagctta cttttaatat
     5461 gaatcttgag agtgaaaatc ttggtgccag aagacttcac ggtgttatgg aaatagtgct
     5521 tgcagatctt ttttttgaag tgcctggcag taagttgaaa aaatttgaaa taaacttgga
     5581 ctatgttaat aaaaaaatac aaattaacga acaaaaagat ttgaattatt atataattta
     5641 gttaaaagca attttaaatg agggaggttt ccatatgtta cgcagcagca acgatgttac
     5701 gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac
     5761 atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg
     5821 tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa
     5881 cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg
     5941 cgctctcgcg gcttacgttc tgcccaggtt tgagcagccg cgtagtgaga tctatatcta
     6001 tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc tcatcaatct
     6061 cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg
     6121 tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt
     6181 tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttacgcgt
//
