LOCUS       sequence-308844-        6542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mreB gRNA expression in B. burgdorferi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6542)
  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 6542)
  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: pBSV2_Psyn-sgRNAmreB
FEATURES             Location/Qualifiers
     source          1..6542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    809..830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(845..850,851..868,869..875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    883..899
                     /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     888..910
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     907..923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     907..923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        960..994
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1016..1091
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1134..1151)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1134..1150)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1143..1165)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1707..2516
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1774..1793)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     regulatory      2767..2776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2773..3147
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3285..3873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3774..3793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaagtaatag agtatgtttt tttgtgtgta tttcttatat gttttattta tgtacgctag
       61 tgttgagata aggactacta atttatactg taatttattg tagcaagttg tggttttact
      121 ggttttgttt atattagata agttttccat ttatagctcc ttgaataccg gattttttat
      181 ctataaagta tatatatcac aattttgatt taaaagtaaa ttcttttttt aaaaatacaa
      241 aaatttttta taaattatat agaccatgat attgtattaa tactaattga ttttaattta
      301 aggttgaact aaaataaaca tagttttgta gaaaataatt tttcattatt ttgctacttg
      361 atattggatc gtaaaaatat tagggcttta caaagttctt ttaaaagaga atttagtaaa
      421 gccctaagtc ttttaataaa aaattttttg taaaaaagtt ggcaaaaata gtttttgcta
      481 tatatttata tataagaaaa aattcaatac tcaaggagat aaaaaatgga aaacttaaac
      541 aataatccac aaaaaattaa tcaaataaat aagcaaacta attttattgg ttttgaaaat
      601 tttatagaac ttgataatca agttaatcaa actaaagaaa aaagtaaatt aagtaagata
      661 ggtaagaaat tacccggaat aagcagtcaa gtctagtgcg gccgcgccca atacgcaaac
      721 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
      781 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      841 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      901 ttcacacagg aaacagctat gaccatgatt acgaattcga gctcgaattc taaagatctt
      961 tgacagctag ctcagtccta ggtataatac tagtgatatc tattgcaaca acagagtttt
     1021 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     1081 cgagtcggtg ctttttttga agcttgggcc cgaacaaaaa ctcaaagctt ggcactggcc
     1141 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1201 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1261 caacagttgc gcagcctgaa tggcgaatgg cgcggccgct acccgagctt caaggaagat
     1321 ttcctattaa ggttgaactt aagagcttaa gcatagacga tttgaaaaaa attttaaaac
     1381 aaacaaaaaa ttctttaata aaacaatatg ttgcgatgtt taaggtttat gatttagatt
     1441 taaagtttag tgaggaagct atagatagaa ttgcagagct tacttttaat atgaatcttg
     1501 agagtgaaaa tcttggtgcc agaagacttc acggtgttat ggaaatagtg cttgcagatc
     1561 ttttttttga agtgcctggc agtaagttga aaaaatttga aataaacttg gactatgtta
     1621 ataaaaaaat acaaattaac gaacaaaaag atttgaatta ttatataatt tagttaaaag
     1681 caattttaaa tgagggaggt ttccatatga gccatattca acgggaaacg tcgaggccgc
     1741 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1801 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1861 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1921 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1981 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2041 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2101 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2161 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2221 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2281 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2341 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2401 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2461 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2521 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgctt
     2581 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatca
     2641 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2701 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atagcacgtg
     2761 aggagggcca ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     2821 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     2881 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     2941 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     3001 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     3061 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     3121 cacttcgtgg ccgaggagca ggactgacac gtgctaaaac ttcattttta atttaaaagg
     3181 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3241 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3301 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3361 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3421 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3481 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3541 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3601 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3661 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3721 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3781 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3841 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3901 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcactaga tccccggcaa
     3961 gttcatcctt agggttttca tatctcttaa aaagtcaatt tctgcttgat taatttcttg
     4021 tggattattg tttttgcggt ttttcatttt ttatctcctt gagtattgaa ttttttctta
     4081 tatataaata tatagcaaaa actatttttg ccaacttttt tacaaaaaat tttttattaa
     4141 aacacttagg gctttgctaa attctctttt aaaagaactt agtaaagccc tatggattta
     4201 agaactgatt atacttacgt agtaaaaata ctacagatat taatagtata gaccatatca
     4261 aaatcataat caatttatat ttatctaaaa ttgtagtcca tacaaaatac aaaaaaatta
     4321 ggtttttcta aattctattt taaaagaact tagtaacacc taatctctaa attacttaca
     4381 aaattctaaa attagatttt tagttcttca tatttcttta aaagtttttt aagaaaatct
     4441 ttttgatttt caaaaatttc atgcatcata aaactagtaa atttggattt ttttttgtaa
     4501 aaatcataac tttcttgggt tttaagttga aatcttaatg gttttattgg attttgccta
     4561 gattttttta acgccggact ttctttatct tttataaaat tataagaatt tttaatacca
     4621 ttttctatta aataatcttc ttttataagc ccctcttgca gtgcagttgc taatcttaaa
     4681 tacttatacg cttgagtttt tgctacatca aagtctttta taaaatcaag aaaagtttta
     4741 taaccatcaa atttataata ttgattatct tttatttctt ttaaaatttt aatattttcc
     4801 accctgtgga atatttcttt tttgaaattg tcatttaact tttcctttag tgtcttgtaa
     4861 taagaaattt tactttcatt aatcattaaa ttttgttcaa cataagtatc tatctttttt
     4921 gttaaaactt taacaacacg ttttttttcc ataaaaattt tcctttattt ttgcaacatt
     4981 ttccttaatt gatataattg ttactatttt agatttttta atatccactt aagaattaat
     5041 ccctcactag aatatttttt attatagaat tcatatacat gtttttctaa atctataaat
     5101 ttatcaacaa ttgctttatt atagggtgta tctatctttt cttttcttaa taatcgtctc
     5161 atattcttta aataacaaca aacactgccg gttttaaacc taaattctat gtaatatatt
     5221 tttgaaaaag tatatacatt ttcaaccccg tttatttcat actttacaaa aacattattt
     5281 attggttttc tataaccata gtaaatgcct aaaaatttgt ccccgtcttt cataggatat
     5341 agataaagtt tttgaatctc tgatttatct aataattttt caaaaaaaat tatacaccgc
     5401 ttttgtttgt tactaactac tctatgcaat gcctttaata tttttgtgtg atatatattt
     5461 ttatcacctt ttttttcttt cagcaggaag atactttctt ttggaaattt aattttttgt
     5521 tcatttattc ttgcagcctt aagtccgatt ttcatattta atccttatat agcctttcta
     5581 tttaattctt ttaaactcaa taaacttttt tggtttttca ttagttctaa taattcgcaa
     5641 taatatgtgc aaaacacttt attgtattct attttctttt ggttattcaa atattctttt
     5701 ataattggct ttagaagttc aatatttact tcttttttta gttgttcaat aaggatattg
     5761 aaaatgtttg tttttaaatc ttgataattt tgttgagaat tttctttttt tctttcaatt
     5821 gacttctcta atttacgctt tatattgttt aaatcgctat atttatgact ttcaataata
     5881 aaatgaggct taaatttgta attttcgtat attttttgga aacttgtttc taattgttta
     5941 gaattatatt catttttttc aaattctttt tgagtgttgc atagaatttc ctttaatttt
     6001 tcttgttttt ctttcataca agatttagta aaatgtatat ttttattttt tagtaattta
     6061 atttctatta tttttatgat tttaagtatt ttgattatat tatctttatc aatatttaaa
     6121 ttgaataaaa ttgtaagaaa aattgaaaga atttttttac ataaaaagtt acatttattg
     6181 aagtaatttc ttacttgata cttttctatt tcattaatct ttctttcttc ttttatatta
     6241 tttttattat ttaaacactc ctttaaatct acactcccat ttttagtaaa tctatcttta
     6301 aggccccttg aaactctatt ttgaaatctt aaatcttttt tttcttgaaa gtatttatag
     6361 attttttggt aacattcttt ttttgaaaaa ttaagttgat agtatatttc ggtccccaaa
     6421 tttaccccta agtgtttgtg ataattagtt gtgactttaa attctttttc taatttgtaa
     6481 aggtaatttt gcagtgtttt tagtttaacg ggtgtttgac tatttctttt aaggttttca
     6541 tt
//
