LOCUS       sequence-292407-        6550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ftsI gRNA expression in B. burgdorferi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6550)
  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 6550)
  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-sgRNAftsI
FEATURES             Location/Qualifiers
     source          1..6550
                     /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    4142..4163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4178..4183,4184..4201,4202..4208)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4216..4232
                     /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     4221..4243
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4240..4256
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4240..4256
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4293..4327
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        4349..4424
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(4467..4484)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4467..4483)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4476..4498)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5040..5849
                     /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(5107..5126)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     regulatory      6103..6112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6109..6483
                     /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"
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 ttagtaaagc cctaagtctt ttaataaaaa attttttgta
     3781 aaaaagttgg caaaaatagt ttttgctata tatttatata taagaaaaaa ttcaatactc
     3841 aaggagataa aaaatggaaa acttaaacaa taatccacaa aaaattaatc aaataaataa
     3901 gcaaactaat tttattggtt ttgaaaattt tatagaactt gataatcaag ttaatcaaac
     3961 taaagaaaaa agtaaattaa gtaagatagg taagaaatta cccggaataa gcagtcaagt
     4021 ctagtctagt gcggccgcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4081 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4141 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     4201 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     4261 attacgaatt cgagctcgaa ttctaaagat ctttgacagc tagctcagtc ctaggtataa
     4321 tactagtgag tgttattaag cattctttgt tttagagcta gaaatagcaa gttaaaataa
     4381 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgaagcttgg
     4441 gcccgaacaa aaactcaaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     4501 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4561 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4621 tggcgcggcc gctacccgag cttcaaggaa gatttcctat taaggttgaa cttaagagct
     4681 taagcataga cgatttgaaa aaaattttaa aacaaacaaa aaattcttta ataaaacaat
     4741 atgttgcgat gtttaaggtt tatgatttag atttaaagtt tagtgaggaa gctatagata
     4801 gaattgcaga gcttactttt aatatgaatc ttgagagtga aaatcttggt gccagaagac
     4861 ttcacggtgt tatggaaata gtgcttgcag atcttttttt tgaagtgcct ggcagtaagt
     4921 tgaaaaaatt tgaaataaac ttggactatg ttaataaaaa aatacaaatt aacgaacaaa
     4981 aagatttgaa ttattatata atttagttaa aagcaatttt aaatgaggga ggtttccata
     5041 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     5101 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     5161 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     5221 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     5281 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     5341 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     5401 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     5461 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     5521 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     5581 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     5641 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     5701 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     5761 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     5821 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     5881 agagcattac gctgacttga cgggacggcg cgcttacaat ttcctgatgc ggtattttct
     5941 ccttacgcat ctgtgcggta tttcacaccg catacaggtg gcacttttcg gggaaatgtg
     6001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6061 caataaccct gataaatgct tcaataatag cacgtgagga gggccaccat ggccaagttg
     6121 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     6181 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     6241 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     6301 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     6361 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     6421 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     6481 tgacacgtgc taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6541 aatctcatga
//
