LOCUS       sequence-311485-        5640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flaB gRNA expression in B. burgdorferi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5640)
  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 5640)
  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_PresTS-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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     13..35
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     32..48
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     32..48
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        197..272
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(290..307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             873..1406
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1509..2097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1998..2017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5574..5595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5610..5615,5616..5633,5634..5640)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat
       61 tcgagctccg aagtttatta ttttatgatt taaattaata aaaagattta tcggtgtgaa
      121 tagaatattt ctttggttta ttatgcatcc tagtacatat tatataattt aatttgaaaa
      181 tttaaatttc tgacttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      241 atcaacttga aaaagtggca ccgagtcggt gctttttttg aagcttggca ctggccgtcg
      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      421 agttgcgcag cctgaatggc gaatggcgcg gccgccctag gttaataccc gagcttcaag
      481 gaagatttcc tattaaggtt gaacttaaga gcttaagcat agacgatttg aaaaaaattt
      541 taaaacaaac aaaaaattct ttaataaaac aatatgttgc gatgtttaag gtttatgatt
      601 tagatttaaa gtttagtgag gaagctatag atagaattgc agagcttact tttaatatga
      661 atcttgagag tgaaaatctt ggtgccagaa gacttcacgg tgttatggaa atagtgcttg
      721 cagatctttt ttttgaagtg cctggcagta agttgaaaaa atttgaaata aacttggact
      781 atgttaataa aaaaatacaa attaacgaac aaaaagattt gaattattat ataatttagt
      841 taaaagcaat tttaaatgag ggaggtttcc atatgttacg cagcagcaac gatgttacgc
      901 agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat
      961 gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg
     1021 agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact
     1081 tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg
     1141 ctctcgcggc ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg
     1201 atctcgcagt ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc
     1261 tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg
     1321 acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg
     1381 atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttacgcgtcc
     1441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1621 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1861 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1921 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1981 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2041 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2101 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2161 tttcctgcac tagatccccg gcaagttcat ccttagggtt ttcatatctc ttaaaaagtc
     2221 aatttctgct tgattaattt cttgtggatt attgtttttg cggtttttca ttttttatct
     2281 ccttgagtat tgaatttttt cttatatata aaatatagca aaaactattt ttgccaactt
     2341 ttttacaaaa aattttttat taaaacactt agggctttgc taaattctct tttaaaagaa
     2401 cttagtaaag ccctatggat ttaagaactg attatactta cgtagtaaaa atactacaga
     2461 tattaatagt atagaccata tcaaaatcat aatcaattta tatttatcta aaattgtggt
     2521 ccatacaaaa tacaaaaaaa ttaggttttt ctaaattcta ttttaaaaga acttagtaac
     2581 acctaatctc taaattactt acaaaattct aaaattagat ttttagttct tcatatttct
     2641 ttaaaagttt tttaagaaaa tctttttgat tttcaaaaat ttcatgcatc ataaaactag
     2701 taaatttgga tttttttttg taaaaatcat aactttcttg ggttttaagt tgaaatctta
     2761 atggttttat tggattttgc ctagattttt ttaacgccgg actttcttta tcttttataa
     2821 aattataaga atttttaata ccattttcta ttaaataatc ttcttttata agcccctctt
     2881 gcagtgcagt tgctaatctt aaatacttat acgcttgagt ttttgctaca tcaaagtctt
     2941 ttataaaatc aagaaaagtt ttataaccat caaatttata atattgatta tcttttattt
     3001 cttttaaaat tttaatattt tccaccctgt ggaatatttc ttttttgaaa ttgtcattta
     3061 acttttcctt tagtgtcttg taataagaaa ttttactttc attaatcatt aaattttgtt
     3121 caacataagt atctatcttt tttgttaaaa ctttaacaac acgttttttt tccataaaaa
     3181 ttttccttta tttttgcaac attttcctta attgatataa ttgttactat tttagatttt
     3241 ttaatatcca cttaagaatt aatccctcac tagaatattt tttattatag aattcatata
     3301 catgtttttc taaatctata aatttatcaa caattgcttt attatagggt gtatctatct
     3361 tttcttttct taataatcgt ctcatattct ttaaataaca acaaacactg ccggttttaa
     3421 acctaaattc tatgtaatat atttttgaaa aagtatatac attttcaacc ccgtttattt
     3481 catactttac aaaaacatta tttattggtt ttctataacc atagtaaatg cctaaaaatt
     3541 tgtccccgtc tttcatagga tatagataaa gtttttgaat ctctgattta tctaataatt
     3601 tttcaaaaaa aattatacac cgcttttgtt tgttactaac tactctatgc aatgccttta
     3661 atatttttgt gtgatatata tttttatcac cttttttttc tttcagcagg aagatacttt
     3721 cttttggaaa tttaattttt tgttcattta ttcttgcagc cttaagtccg attttcatat
     3781 ttaatcctta tatagccttt ctatttaatt cttttaaact caataaactt ttttggtttt
     3841 tcattagttc taataattcg caataatatg tgcaaaacac tttattgtat tctattttct
     3901 tttggttatt caaatattct tttataattg gctttagaag ttcaatattt acttcttttt
     3961 ttagttgttc aataaggata ttgaaaatgt ttgtttttaa atcttgataa ttttgttgag
     4021 aattttcttt ttttctttca attgacttct ctaatttacg ctttatattg tttaaatcgc
     4081 tatatttatg actttcaata ataaaatgag gcttaaattt gtaattttcg tatatttttt
     4141 ggaaacttgt ttctaattgt ttagaattat attcattttt ttcaaattct ttttgagtgt
     4201 tgcatagaat ttcctttaat ttttcttgtt tttctttcat acaagattta gtaaaatgta
     4261 tatttttatt ttttagtaat ttaatttcta ttatttttat gattttaagt attttgatta
     4321 tattatcttt atcaatattt aaattgaata aaattgtaag aaaaattgaa agaatttttt
     4381 tacataaaaa gttacattta ttgaagtaat ttcttacttg atacttttct atttcattaa
     4441 tctttctttc ttcttttata ttatttttat tatttaaaca ctcctttaaa tctacactcc
     4501 catttttagt aaatctatct ttaaggcccc ttgaaactct attttgaaat cttaaatctt
     4561 ttttttcttg aaagtattta tagatttttt ggtaacattc tttttttgaa aaattaagtt
     4621 gatagtatat ttcggtcccc aaatttaccc ctaagtgttt gtgataatta gttgtgactt
     4681 taaattcttt ttctaatttg taaaggtaat tttgcagtgt ttttagttta acgggtgttt
     4741 gactatttct tttaaggttt tcattaaagt aatagagtat gtttttttgt gtgtatttct
     4801 tatatgtttt atttatgtac gctagtgttg agataaggac tactaattta tactgtaatt
     4861 tattgtagca agttgtggtt ttactggttt tgtttatatt agataagttt tccatttata
     4921 gctccttgaa taccggattt tttatctata aagtatatat atcacaattt tgatttaaaa
     4981 gtaaattctt tttttaaaaa tacaaaaatt ttttataaat tatatagacc atgatattgt
     5041 attaatacta attgatttta atttaaggtt gaactaaaat aaacatagtt ttgtagaaaa
     5101 taatttttca ttattttgct acttgatatt ggatcgtaaa aatattaggg ctttacaaag
     5161 ttcttttaaa agagaattta gtaaagccct aagtctttta ataaaaaatt ttttgtaaaa
     5221 aagttggcaa aaatagtttt tgctatatat ttatatataa gaaaaaattc aatactcaag
     5281 gagataaaaa atggaaaact taaacaataa tccacaaaaa attaatcaaa taaataagca
     5341 aactaatttt attggttttg aaaattttat agaacttgat aatcaagtta atcaaactaa
     5401 agaaaaaagt aaattaagta agataggtaa gaaattaccc ggaataagca gtcaagtcta
     5461 gtgcggccgc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5521 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     5581 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
//
