LOCUS       sequence-309487-        5608 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 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-sgRNAflaB
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..33,34..40)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    48..64
                     /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     53..75
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     72..88
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     72..88
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        124..158
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        180..255
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(298..315)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(298..314)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(307..329)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             881..1414
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1517..2105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2006..2025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5582..5603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
       61 acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcga attctaaaga
      121 tctttgacag ctagctcagt cctaggtata atactagtga aaatttaaat ttctgacttg
      181 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      241 gcaccgagtc ggtgcttttt ttgaagcttg ggcccgaaca aaaactcaaa gcttggcact
      301 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      361 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      421 ttcccaacag ttgcgcagcc tgaatggcga atggcgcggc cgccctaggt taatacccga
      481 gcttcaagga agatttccta ttaaggttga acttaagagc ttaagcatag acgatttgaa
      541 aaaaatttta aaacaaacaa aaaattcttt aataaaacaa tatgttgcga tgtttaaggt
      601 ttatgattta gatttaaagt ttagtgagga agctatagat agaattgcag agcttacttt
      661 taatatgaat cttgagagtg aaaatcttgg tgccagaaga cttcacggtg ttatggaaat
      721 agtgcttgca gatctttttt ttgaagtgcc tggcagtaag ttgaaaaaat ttgaaataaa
      781 cttggactat gttaataaaa aaatacaaat taacgaacaa aaagatttga attattatat
      841 aatttagtta aaagcaattt taaatgaggg aggtttccat atgttacgca gcagcaacga
      901 tgttacgcag cagggcagtc gccctaaaac aaagttaggt ggctcaagta tgggcatcat
      961 tcgcacatgt aggctcggcc ctgaccaagt caaatccatg cgggctgctc ttgatctttt
     1021 cggtcgtgag ttcggagacg tagccaccta ctcccaacat cagccggact ccgattacct
     1081 cgggaacttg ctccgtagta agacattcat cgcgcttgct gccttcgacc aagaagcggt
     1141 tgttggcgct ctcgcggctt acgttctgcc caagtttgag cagccgcgta gtgagatcta
     1201 tatctatgat ctcgcagtct ccggcgagca ccggaggcag ggcattgcca ccgcgctcat
     1261 caatctcctc aagcatgagg ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga
     1321 ttacggtgac gatcccgcag tggctctcta tacaaagttg ggcatacggg aagaagtgat
     1381 gcactttgat atcgacccaa gtaccgccac ctaacaattc gttcaagccg agatcggctt
     1441 acgcgtccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1501 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1561 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1621 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1681 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1741 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1801 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1861 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1921 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1981 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2041 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2101 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2161 acatgttctt tcctgcacta gatccccggc aagttcatcc ttagggtttt catatctctt
     2221 aaaaagtcaa tttctgcttg attaatttct tgtggattat tgtttttgcg gtttttcatt
     2281 ttttatctcc ttgagtattg aattttttct tatatataaa atatagcaaa aactattttt
     2341 gccaactttt ttacaaaaaa ttttttatta aaacacttag ggctttgcta aattctcttt
     2401 taaaagaact tagtaaagcc ctatggattt aagaactgat tatacttacg tagtaaaaat
     2461 actacagata ttaatagtat agaccatatc aaaatcataa tcaatttata tttatctaaa
     2521 attgtggtcc atacaaaata caaaaaaatt aggtttttct aaattctatt ttaaaagaac
     2581 ttagtaacac ctaatctcta aattacttac aaaattctaa aattagattt ttagttcttc
     2641 atatttcttt aaaagttttt taagaaaatc tttttgattt tcaaaaattt catgcatcat
     2701 aaaactagta aatttggatt tttttttgta aaaatcataa ctttcttggg ttttaagttg
     2761 aaatcttaat ggttttattg gattttgcct agattttttt aacgccggac tttctttatc
     2821 ttttataaaa ttataagaat ttttaatacc attttctatt aaataatctt cttttataag
     2881 cccctcttgc agtgcagttg ctaatcttaa atacttatac gcttgagttt ttgctacatc
     2941 aaagtctttt ataaaatcaa gaaaagtttt ataaccatca aatttataat attgattatc
     3001 ttttatttct tttaaaattt taatattttc caccctgtgg aatatttctt ttttgaaatt
     3061 gtcatttaac ttttccttta gtgtcttgta ataagaaatt ttactttcat taatcattaa
     3121 attttgttca acataagtat ctatcttttt tgttaaaact ttaacaacac gttttttttc
     3181 cataaaaatt ttcctttatt tttgcaacat tttccttaat tgatataatt gttactattt
     3241 tagatttttt aatatccact taagaattaa tccctcacta gaatattttt tattatagaa
     3301 ttcatataca tgtttttcta aatctataaa tttatcaaca attgctttat tatagggtgt
     3361 atctatcttt tcttttctta ataatcgtct catattcttt aaataacaac aaacactgcc
     3421 ggttttaaac ctaaattcta tgtaatatat ttttgaaaaa gtatatacat tttcaacccc
     3481 gtttatttca tactttacaa aaacattatt tattggtttt ctataaccat agtaaatgcc
     3541 taaaaatttg tccccgtctt tcataggata tagataaagt ttttgaatct ctgatttatc
     3601 taataatttt tcaaaaaaaa ttatacaccg cttttgtttg ttactaacta ctctatgcaa
     3661 tgcctttaat atttttgtgt gatatatatt tttatcacct tttttttctt tcagcaggaa
     3721 gatactttct tttggaaatt taattttttg ttcatttatt cttgcagcct taagtccgat
     3781 tttcatattt aatccttata tagcctttct atttaattct tttaaactca ataaactttt
     3841 ttggtttttc attagttcta ataattcgca ataatatgtg caaaacactt tattgtattc
     3901 tattttcttt tggttattca aatattcttt tataattggc tttagaagtt caatatttac
     3961 ttcttttttt agttgttcaa taaggatatt gaaaatgttt gtttttaaat cttgataatt
     4021 ttgttgagaa ttttcttttt ttctttcaat tgacttctct aatttacgct ttatattgtt
     4081 taaatcgcta tatttatgac tttcaataat aaaatgaggc ttaaatttgt aattttcgta
     4141 tattttttgg aaacttgttt ctaattgttt agaattatat tcattttttt caaattcttt
     4201 ttgagtgttg catagaattt cctttaattt ttcttgtttt tctttcatac aagatttagt
     4261 aaaatgtata tttttatttt ttagtaattt aatttctatt atttttatga ttttaagtat
     4321 tttgattata ttatctttat caatatttaa attgaataaa attgtaagaa aaattgaaag
     4381 aattttttta cataaaaagt tacatttatt gaagtaattt cttacttgat acttttctat
     4441 ttcattaatc tttctttctt cttttatatt atttttatta tttaaacact cctttaaatc
     4501 tacactccca tttttagtaa atctatcttt aaggcccctt gaaactctat tttgaaatct
     4561 taaatctttt ttttcttgaa agtatttata gattttttgg taacattctt tttttgaaaa
     4621 attaagttga tagtatattt cggtccccaa atttacccct aagtgtttgt gataattagt
     4681 tgtgacttta aattcttttt ctaatttgta aaggtaattt tgcagtgttt ttagtttaac
     4741 gggtgtttga ctatttcttt taaggttttc attaaagtaa tagagtatgt ttttttgtgt
     4801 gtatttctta tatgttttat ttatgtacgc tagtgttgag ataaggacta ctaatttata
     4861 ctgtaattta ttgtagcaag ttgtggtttt actggttttg tttatattag ataagttttc
     4921 catttatagc tccttgaata ccggattttt tatctataaa gtatatatat cacaattttg
     4981 atttaaaagt aaattctttt tttaaaaata caaaaatttt ttataaatta tatagaccat
     5041 gatattgtat taatactaat tgattttaat ttaaggttga actaaaataa acatagtttt
     5101 gtagaaaata atttttcatt attttgctac ttgatattgg atcgtaaaaa tattagggct
     5161 ttacaaagtt cttttaaaag agaatttagt aaagccctaa gtcttttaat aaaaaatttt
     5221 ttgtaaaaaa gttggcaaaa atagtttttg ctatatattt atatataaga aaaaattcaa
     5281 tactcaagga gataaaaaat ggaaaactta aacaataatc cacaaaaaat taatcaaata
     5341 aataagcaaa ctaattttat tggttttgaa aattttatag aacttgataa tcaagttaat
     5401 caaactaaag aaaaaagtaa attaagtaag ataggtaaga aattacccgg aataagcagt
     5461 caagtctagt gcggccgcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5521 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5581 ttaatgtgag ttagctcact cattaggc
//
