LOCUS       sequence-311460-        6104 bp    DNA     circular SYN 15-APR-2025
DEFINITION  gRNA expression in B. burgdorferi, parental vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6104)
  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 6104)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-sgRNA500
FEATURES             Location/Qualifiers
     source          1..6104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..24
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     21..37
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     21..37
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        74..108
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        625..700
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(743..760)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(743..759)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(752..774)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1326..1859
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      1962..2550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2451..2470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6027..6048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6063..6068,6069..6086,6087..6093)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(6101..6104,1..13)
                     /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)."
ORIGIN
        1 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat tcgagctcga
       61 attctaaaga tctttgacag ctagctcagt cctaggtata atactagtgg aagagccaaa
      121 gtcaacactt atagacaaat atgatctttc taaccttcac gaaatagcat cagggggagc
      181 acctttgtct aaggaagtag gagaggcagt tgcaaagagg tttcatttac ccggaattag
      241 gcaaggatat gggttaacag aaaccacatc tgcaatttta attacacctg aaggagatga
      301 taaaccagga gctgttggga aggttgttcc tttttttgag gctaaagttg ttgatttaga
      361 tactggaaag actttaggag ttaatcaaag aggcgagtta tgtgtaagag gacctatgat
      421 aatgtcagga tacgtaaata atccagaagc tacaaatgca ttgatagata aagatggttg
      481 gttacactca ggcgatatag cttattggga cgaagatgaa cactttttta tagttgatag
      541 attgaaatct ttgattaagt ataaaggtta tcaagtagct ccagcagaat tggaaagtat
      601 tttattgcaa cacccagctc ttccgtttta gagctagaaa tagcaagtta aaataaggct
      661 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgaa gcttgggccc
      721 gaacaaaaac tcaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      781 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      841 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      901 gcggccgccc taggttaata cccgagcttc aaggaagatt tcctattaag gttgaactta
      961 agagcttaag catagacgat ttgaaaaaaa ttttaaaaca aacaaaaaat tctttaataa
     1021 aacaatatgt tgcgatgttt aaggtttatg atttagattt aaagtttagt gaggaagcta
     1081 tagatagaat tgcagagctt acttttaata tgaatcttga gagtgaaaat cttggtgcca
     1141 gaagacttca cggtgttatg gaaatagtgc ttgcagatct tttttttgaa gtgcctggca
     1201 gtaagttgaa aaaatttgaa ataaacttgg actatgttaa taaaaaaata caaattaacg
     1261 aacaaaaaga tttgaattat tatataattt agttaaaagc aattttaaat gagggaggtt
     1321 tccatatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     1381 taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat
     1441 ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc
     1501 aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc
     1561 ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt
     1621 ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga
     1681 ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg
     1741 cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa
     1801 agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac
     1861 aattcgttca agccgagatc ggcttacgcg tccaaaatcc cttaacgtga gttttcgttc
     1921 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1981 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2041 gatcaagagc taccaactct ttttccgaag gtaactggat tcagcagagc gcagatacca
     2101 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2161 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2221 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2281 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2341 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2401 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2461 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2521 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2581 ctggcctttt gctggccttt tgctcacatg ttctttcctg cactagatcc ccggcaagtt
     2641 catccttagg gttttcatat ctcttaaaaa gtcaatttct gcttgattaa tttcttgtgg
     2701 attattgttt ttgcggtttt tcatttttta tctccttgag tattgaattt tttcttatat
     2761 ataaaatata gcaaaaacta tttttgccaa cttttttaca aaaaattttt tattaaaaca
     2821 cttagggctt tgctaaattc tcttttaaaa gaacttagta aagccctatg gatttaagaa
     2881 ctgattatac ttacgtagta aaaatactac agatattaat agtatagacc atatcaaaat
     2941 cataatcaat ttatatttat ctaaaattgt ggtccataca aaatacaaaa aaattaggtt
     3001 tttctaaatt ctattttaaa agaacttagt aacacctaat ctctaaatta cttacaaaat
     3061 tctaaaatta gatttttagt tcttcatatt tctttaaaag ttttttaaga aaatcttttt
     3121 gattttcaaa aatttcatgc atcataaaac tagtaaattt ggattttttt ttgtaaaaat
     3181 cataactttc ttgggtttta agttgaaatc ttaatggttt tattggattt tgcctagatt
     3241 tttttaacgc cggactttct ttatctttta taaaattata agaattttta ataccatttt
     3301 ctattaaata atcttctttt ataagcccct cttgcagtgc agttgctaat cttaaatact
     3361 tatacgcttg agtttttgct acatcaaagt cttttataaa atcaagaaaa gttttataac
     3421 catcaaattt ataatattga ttatctttta tttcttttaa aattttaata ttttccaccc
     3481 tgtggaatat ttcttttttg aaattgtcat ttaacttttc ctttagtgtc ttgtaataag
     3541 aaattttact ttcattaatc attaaatttt gttcaacata agtatctatc ttttttgtta
     3601 aaactttaac aacacgtttt ttttccataa aaattttcct ttatttttgc aacattttcc
     3661 ttaattgata taattgttac tattttagat tttttaatat ccacttaaga attaatccct
     3721 cactagaata ttttttatta tagaattcat atacatgttt ttctaaatct ataaatttat
     3781 caacaattgc tttattatag ggtgtatcta tcttttcttt tcttaataat cgtctcatat
     3841 tctttaaata acaacaaaca ctgccggttt taaacctaaa ttctatgtaa tatatttttg
     3901 aaaaagtata tacattttca accccgttta tttcatactt tacaaaaaca ttatttattg
     3961 gttttctata accatagtaa atgcctaaaa atttgtcccc gtctttcata ggatatagat
     4021 aaagtttttg aatctctgat ttatctaata atttttcaaa aaaaattata caccgctttt
     4081 gtttgttact aactactcta tgcaatgcct ttaatatttt tgtgtgatat atatttttat
     4141 cacctttttt ttctttcagc aggaagatac tttcttttgg aaatttaatt ttttgttcat
     4201 ttattcttgc agccttaagt ccgattttca tatttaatcc ttatatagcc tttctattta
     4261 attcttttaa actcaataaa cttttttggt ttttcattag ttctaataat tcgcaataat
     4321 atgtgcaaaa cactttattg tattctattt tcttttggtt attcaaatat tcttttataa
     4381 ttggctttag aagttcaata tttacttctt tttttagttg ttcaataagg atattgaaaa
     4441 tgtttgtttt taaatcttga taattttgtt gagaattttc tttttttctt tcaattgact
     4501 tctctaattt acgctttata ttgtttaaat cgctatattt atgactttca ataataaaat
     4561 gaggcttaaa tttgtaattt tcgtatattt tttggaaact tgtttctaat tgtttagaat
     4621 tatattcatt tttttcaaat tctttttgag tgttgcatag aatttccttt aatttttctt
     4681 gtttttcttt catacaagat ttagtaaaat gtatattttt attttttagt aatttaattt
     4741 ctattatttt tatgatttta agtattttga ttatattatc tttatcaata tttaaattga
     4801 ataaaattgt aagaaaaatt gaaagaattt ttttacataa aaagttacat ttattgaagt
     4861 aatttcttac ttgatacttt tctatttcat taatctttct ttcttctttt atattatttt
     4921 tattatttaa acactccttt aaatctacac tcccattttt agtaaatcta tctttaaggc
     4981 cccttgaaac tctattttga aatcttaaat cttttttttc ttgaaagtat ttatagattt
     5041 tttggtaaca ttcttttttt gaaaaattaa gttgatagta tatttcggtc cccaaattta
     5101 cccctaagtg tttgtgataa ttagttgtga ctttaaattc tttttctaat ttgtaaaggt
     5161 aattttgcag tgtttttagt ttaacgggtg tttgactatt tcttttaagg ttttcattaa
     5221 agtaatagag tatgtttttt tgtgtgtatt tcttatatgt tttatttatg tacgctagtg
     5281 ttgagataag gactactaat ttatactgta atttattgta gcaagttgtg gttttactgg
     5341 ttttgtttat attagataag ttttccattt atagctcctt gaataccgga ttttttatct
     5401 ataaagtata tatatcacaa ttttgattta aaagtaaatt ctttttttaa aaatacaaaa
     5461 attttttata aattatatag accatgatat tgtattaata ctaattgatt ttaatttaag
     5521 gttgaactaa aataaacata gttttgtaga aaataatttt tcattatttt gctacttgat
     5581 attggatcgt aaaaatatta gggctttaca aagttctttt aaaagagaat ttagtaaagc
     5641 cctaagtctt ttaataaaaa attttttgta aaaaagttgg caaaaatagt ttttgctata
     5701 tatttatata taagaaaaaa ttcaatactc aagtagataa aaaatggaaa acttaaacaa
     5761 taatccacaa aaaattaatc aaataaataa gcaaactaat tttattggtt ttgaaaattt
     5821 tatagaactt gataatcaag ttaatcaaac taaagaaaaa agtaaattaa gtaagatagg
     5881 taagaaatta cccggaataa gcagtcaagt ctagtgcggc cgcgcccaat acgcaaaccg
     5941 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6001 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6061 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgt
//
