LOCUS       sequence-309491-        5608 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 5608)
  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 5608)
  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_Psyn-sgRNAmreB
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1703..1724
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1739..1744,1745..1762,1763..1769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1777..1793
                     /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     1782..1804
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1801..1817
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1801..1817
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1853..1887
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1909..1984
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2027..2044)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2027..2043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2036..2058)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2610..3143
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      3246..3834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3735..3754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaaaacact ttattgtatt ctattttctt ttggttattc aaatattctt ttataattgg
       61 ctttagaagt tcaatattta cttctttttt tagttgttca ataaggatat tgaaaatgtt
      121 tgtttttaaa tcttgataat tttgttgaga attttctttt tttctttcaa ttgacttctc
      181 taatttacgc tttatattgt ttaaatcgct atatttatga ctttcaataa taaaatgagg
      241 cttaaatttg taattttcgt atattttttg gaaacttgtt tctaattgtt tagaattata
      301 ttcatttttt tcaaattctt tttgagtgtt gcatagaatt tcctttaatt tttcttgttt
      361 ttctttcata caagatttag taaaatgtat atttttattt tttagtaatt taatttctat
      421 tatttttatg attttaagta ttttgattat attatcttta tcaatattta aattgaataa
      481 aattgtaaga aaaattgaaa gaattttttt acataaaaag ttacatttat tgaagtaatt
      541 tcttacttga tacttttcta tttcattaat ctttctttct tcttttatat tatttttatt
      601 atttaaacac tcctttaaat ctacactccc atttttagta aatctatctt taaggcccct
      661 tgaaactcta ttttgaaatc ttaaatcttt tttttcttga aagtatttat agattttttg
      721 gtaacattct ttttttgaaa aattaagttg atagtatatt tcggtcccca aatttacccc
      781 taagtgtttg tgataattag ttgtgacttt aaattctttt tctaatttgt aaaggtaatt
      841 ttgcagtgtt tttagtttaa cgggtgtttg actatttctt ttaaggtttt cattaaagta
      901 atagagtatg tttttttgtg tgtatttctt atatgtttta tttatgtacg ctagtgttga
      961 gataaggact actaatttat actgtaattt attgtagcaa gttgtggttt tactggtttt
     1021 gtttatatta gataagtttt ccatttatag ctccttgaat accggatttt ttatctataa
     1081 agtatatata tcacaatttt gatttaaaag taaattcttt ttttaaaaat acaaaaattt
     1141 tttataaatt atatagacca tgatattgta ttaatactaa ttgattttaa tttaaggttg
     1201 aactaaaata aacatagttt tgtagaaaat aatttttcat tattttgcta cttgatattg
     1261 gatcgtaaaa atattagggc tttacaaagt tcttttaaaa gagaatttag taaagcccta
     1321 agtcttttaa taaaaaattt tttgtaaaaa agttggcaaa aatagttttt gctatatatt
     1381 tatatataag aaaaaattca atactcaagg agataaaaaa tggaaaactt aaacaataat
     1441 ccacaaaaaa ttaatcaaat aaataagcaa actaatttta ttggttttga aaattttata
     1501 gaacttgata atcaagttaa tcaaactaaa gaaaaaagta aattaagtaa gataggtaag
     1561 aaattacccg gaataagcag tcaagtctag tgcggccgcg cccaatacgc aaaccgcctc
     1621 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     1681 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     1741 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     1801 caggaaacag ctatgacatg attacgaatt cgagctcgaa ttctaaagat ctttgacagc
     1861 tagctcagtc ctaggtataa tactagtgat atctattgca acaacagagt tttagagcta
     1921 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     1981 gtgctttttt tgaagcttgg gcccgaacaa aaactcaaag cttggcactg gccgtcgttt
     2041 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2101 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2161 tgcgcagcct gaatggcgaa tggcgcggcc gccctaggtt aatacccgag cttcaaggaa
     2221 gatttcctat taaggttgaa cttaagagct taagcataga cgatttgaaa aaaattttaa
     2281 aacaaacaaa aaattcttta ataaaacaat atgttgcgat gtttaaggtt tatgatttag
     2341 atttaaagtt tagtgaggaa gctatagata gaattgcaga gcttactttt aatatgaatc
     2401 ttgagagtga aaatcttggt gccagaagac ttcacggtgt tatggaaata gtgcttgcag
     2461 atcttttttt tgaagtgcct ggcagtaagt tgaaaaaatt tgaaataaac ttggactatg
     2521 ttaataaaaa aatacaaatt aacgaacaaa aagatttgaa ttattatata atttagttaa
     2581 aagcaatttt aaatgaggga ggtttccata tgttacgcag cagcaacgat gttacgcagc
     2641 agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta
     2701 ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt
     2761 tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc gggaacttgc
     2821 tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt gttggcgctc
     2881 tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat atctatgatc
     2941 tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc aatctcctca
     3001 agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg
     3061 atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata
     3121 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggctta cgcgtccaaa
     3181 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3361 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3421 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3481 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3541 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3601 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3661 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3721 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3781 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3841 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3901 cctgcactag atccccggca agttcatcct tagggttttc atatctctta aaaagtcaat
     3961 ttctgcttga ttaatttctt gtggattatt gtttttgcgg tttttcattt tttatctcct
     4021 tgagtattga attttttctt atatataaaa tatagcaaaa actatttttg ccaacttttt
     4081 tacaaaaaat tttttattaa aacacttagg gctttgctaa attctctttt aaaagaactt
     4141 agtaaagccc tatggattta agaactgatt atacttacgt agtaaaaata ctacagatat
     4201 taatagtata gaccatatca aaatcataat caatttatat ttatctaaaa ttgtggtcca
     4261 tacaaaatac aaaaaaatta ggtttttcta aattctattt taaaagaact tagtaacacc
     4321 taatctctaa attacttaca aaattctaaa attagatttt tagttcttca tatttcttta
     4381 aaagtttttt aagaaaatct ttttgatttt caaaaatttc atgcatcata aaactagtaa
     4441 atttggattt ttttttgtaa aaatcataac tttcttgggt tttaagttga aatcttaatg
     4501 gttttattgg attttgccta gattttttta acgccggact ttctttatct tttataaaat
     4561 tataagaatt tttaatacca ttttctatta aataatcttc ttttataagc ccctcttgca
     4621 gtgcagttgc taatcttaaa tacttatacg cttgagtttt tgctacatca aagtctttta
     4681 taaaatcaag aaaagtttta taaccatcaa atttataata ttgattatct tttatttctt
     4741 ttaaaatttt aatattttcc accctgtgga atatttcttt tttgaaattg tcatttaact
     4801 tttcctttag tgtcttgtaa taagaaattt tactttcatt aatcattaaa ttttgttcaa
     4861 cataagtatc tatctttttt gttaaaactt taacaacacg ttttttttcc ataaaaattt
     4921 tcctttattt ttgcaacatt ttccttaatt gatataattg ttactatttt agatttttta
     4981 atatccactt aagaattaat ccctcactag aatatttttt attatagaat tcatatacat
     5041 gtttttctaa atctataaat ttatcaacaa ttgctttatt atagggtgta tctatctttt
     5101 cttttcttaa taatcgtctc atattcttta aataacaaca aacactgccg gttttaaacc
     5161 taaattctat gtaatatatt tttgaaaaag tatatacatt ttcaaccccg tttatttcat
     5221 actttacaaa aacattattt attggttttc tataaccata gtaaatgcct aaaaatttgt
     5281 ccccgtcttt cataggatat agataaagtt tttgaatctc tgatttatct aataattttt
     5341 caaaaaaaat tatacaccgc ttttgtttgt tactaactac tctatgcaat gcctttaata
     5401 tttttgtgtg atatatattt ttatcacctt ttttttcttt cagcaggaag atactttctt
     5461 ttggaaattt aattttttgt tcatttattc ttgcagcctt aagtccgatt ttcatattta
     5521 atccttatat agcctttcta tttaattctt ttaaactcaa taaacttttt tggtttttca
     5581 ttagttctaa taattcgcaa taatatgt
//
