LOCUS       sequence-234734-        8730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty E. coli - B. burgdorferi shuttle vector; hygromycin B 
            resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8730)
  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 8730)
  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: pBSV2H
FEATURES             Location/Qualifiers
     source          1..8730
                     /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    4136..4157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4172..4177,4178..4195,4196..4202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4210..4226
                     /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     4215..4237
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4234..4250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4234..4250
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4353..4370)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4353..4369)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4362..4384)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5576..6595
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             7679..8698
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
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 atactgttct 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 tggccttttg ctggcctttt gctcacatgt
      721 tctttcctgc actagatccc cggcaagttc atccttaggg ttttcatatc tcttaaaaag
      781 tcaatttctg cttgattaat ttcttgtgga ttattgtttt tgcggttttt cattttttat
      841 ctccttgagt attgaatttt ttcttatata taaaatatag caaaaactat ttttgccaac
      901 ttttttacaa aaaatttttt attaaaacac ttagggcttt gctaaattct cttttaaaag
      961 aacttagtaa agccctatgg atttaagaac tgattatact tacgtagtaa aaatactaca
     1021 gatattaata gtatagacca tatcaaaatc ataatcaatt tatatttatc taaaattgtg
     1081 gtccatacaa aatacaaaaa aattaggttt ttctaaattc tattttaaaa gaacttagta
     1141 acacctaatc tctaaattac ttacaaaatt ctaaaattag atttttagtt cttcatattt
     1201 ctttaaaagt tttttaagaa aatctttttg attttcaaaa atttcatgca tcataaaact
     1261 agtaaatttg gatttttttt tgtaaaaatc ataactttct tgggttttaa gttgaaatct
     1321 taatggtttt attggatttt gcctagattt ttttaacgcc ggactttctt tatcttttat
     1381 aaaattataa gaatttttaa taccattttc tattaaataa tcttctttta taagcccctc
     1441 ttgcagtgca gttgctaatc ttaaatactt atacgcttga gtttttgcta catcaaagtc
     1501 ttttataaaa tcaagaaaag ttttataacc atcaaattta taatattgat tatcttttat
     1561 ttcttttaaa attttaatat tttccaccct gtggaatatt tcttttttga aattgtcatt
     1621 taacttttcc tttagtgtct tgtaataaga aattttactt tcattaatca ttaaattttg
     1681 ttcaacataa gtatctatct tttttgttaa aactttaaca acacgttttt tttccataaa
     1741 aattttcctt tatttttgca acattttcct taattgatat aattgttact attttagatt
     1801 ttttaatatc cacttaagaa ttaatccctc actagaatat tttttattat agaattcata
     1861 tacatgtttt tctaaatcta taaatttatc aacaattgct ttattatagg gtgtatctat
     1921 cttttctttt cttaataatc gtctcatatt ctttaaataa caacaaacac tgccggtttt
     1981 aaacctaaat tctatgtaat atatttttga aaaagtatat acattttcaa ccccgtttat
     2041 ttcatacttt acaaaaacat tatttattgg ttttctataa ccatagtaaa tgcctaaaaa
     2101 tttgtccccg tctttcatag gatatagata aagtttttga atctctgatt tatctaataa
     2161 tttttcaaaa aaaattatac accgcttttg tttgttacta actactctat gcaatgcctt
     2221 taatattttt gtgtgatata tatttttatc accttttttt tctttcagca ggaagatact
     2281 ttcttttgga aatttaattt tttgttcatt tattcttgca gccttaagtc cgattttcat
     2341 atttaatcct tatatagcct ttctatttaa ttcttttaaa ctcaataaac ttttttggtt
     2401 tttcattagt tctaataatt cgcaataata tgtgcaaaac actttattgt attctatttt
     2461 cttttggtta ttcaaatatt cttttataat tggctttaga agttcaatat ttacttcttt
     2521 ttttagttgt tcaataagga tattgaaaat gtttgttttt aaatcttgat aattttgttg
     2581 agaattttct ttttttcttt caattgactt ctctaattta cgctttatat tgtttaaatc
     2641 gctatattta tgactttcaa taataaaatg aggcttaaat ttgtaatttt cgtatatttt
     2701 ttggaaactt gtttctaatt gtttagaatt atattcattt ttttcaaatt ctttttgagt
     2761 gttgcataga atttccttta atttttcttg tttttctttc atacaagatt tagtaaaatg
     2821 tatattttta ttttttagta atttaatttc tattattttt atgattttaa gtattttgat
     2881 tatattatct ttatcaatat ttaaattgaa taaaattgta agaaaaattg aaagaatttt
     2941 tttacataaa aagttacatt tattgaagta atttcttact tgatactttt ctatttcatt
     3001 aatctttctt tcttctttta tattattttt attatttaaa cactccttta aatctacact
     3061 cccattttta gtaaatctat ctttaaggcc ccttgaaact ctattttgaa atcttaaatc
     3121 ttttttttct tgaaagtatt tatagatttt ttggtaacat tctttttttg aaaaattaag
     3181 ttgatagtat atttcggtcc ccaaatttac ccctaagtgt ttgtgataat tagttgtgac
     3241 tttaaattct ttttctaatt tgtaaaggta attttgcagt gtttttagtt taacgggtgt
     3301 ttgactattt cttttaaggt tttcattaaa gtaatagagt atgttttttt gtgtgtattt
     3361 cttatatgtt ttatttatgt acgctagtgt tgagataagg actactaatt tatactgtaa
     3421 tttattgtag caagttgtgg ttttactggt tttgtttata ttagataagt tttccattta
     3481 tagctccttg aataccggat tttttatcta taaagtatat atatcacaat tttgatttaa
     3541 aagtaaattc tttttttaaa aatacaaaaa ttttttataa attatataga ccatgatatt
     3601 gtattaatac taattgattt taatttaagg ttgaactaaa ataaacatag ttttgtagaa
     3661 aataattttt cattattttg ctacttgata ttggatcgta aaaatattag ggctttacaa
     3721 agttctttta aaagagaatt tagtaaagcc ctaagtcttt taataaaaaa ttttttgtaa
     3781 aaaagttggc aaaaatagtt tttgctatat atttatatat aagaaaaaat tcaatactca
     3841 aggagataaa aaatggaaaa cttaaacaat aatccacaaa aaattaatca aataaataag
     3901 caaactaatt ttattggttt tgaaaatttt atagaacttg ataatcaagt taatcaaact
     3961 aaagaaaaaa gtaaattaag taagataggt aagaaattac ccggaataag cagtcaagtc
     4021 tagtgcggcc gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4081 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4141 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4201 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
     4261 attcgagctc ggtacaccgg tctcgaggac gtcgctagcg gatcccgggg taccgatcct
     4321 ctagagtcga cctgcaggca tgcaagcttg gcactggccg tcgttttaca acgtcgtgac
     4381 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4441 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4501 ggcgaatggc gcggccgccc taggttaatc ttcaataaca tgtaaaccac gacgttttaa
     4561 atcttctaat gaatctaaca ttcctcttat taattcaaca ggatgtcctt cccaatcttc
     4621 aacaacacca acaattctta aaggttcaca agttctataa gattgtgttg gattaccagg
     4681 aaatttttta tttgtaagat ttggatcatc ttcaaatggt cctgttggtt caactatata
     4741 tatataacca cgaccttcaa gaccagatag tgacatagca agttcagctc cccaaactgc
     4801 tggttccatt aaagctgaaa aataaatatg tttaagaatt cgaccatctt caaaatgaga
     4861 aataaatcct gtagttagta aatcaccaat tgctaaatta gctttagttc catgataaaa
     4921 tggtccttgt acttgtttat aattatcatg agaaatagga atccaatctt ttaccatgac
     4981 gttacctcct catgatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt
     5041 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
     5101 gaggcataga ctgtacccca aaaaaacagt cataacaagc catgcggccg tacccgagct
     5161 tcaaggaaga tttcctatta aggttgaact taagagctta agcatagacg atttgaaaaa
     5221 aattttaaaa caaacaaaaa attctttaat aaaacaatat gttgcgatgt ttaaggttta
     5281 tgatttagat ttaaagttta gtgaggaagc tatagataga attgcagagc ttacttttaa
     5341 tatgaatctt gagagtgaaa atcttggtgc cagaagactt cacggtgtta tggaaatagt
     5401 gcttgcagat cttttttttg aagtgcctgg cagtaagttg aaaaaatttg aaataaactt
     5461 ggactatgtt aataaaaaaa tacaaattaa cgaacaaaaa gatttgaatt attatataat
     5521 ttagttaaaa gcaattttaa atgagggagg tttccatcat atggatagaa gtggtaaacc
     5581 tgaattaact gctactagtg ttgaaaaatt tttaattgaa aaatttgata gtgttagtga
     5641 tttaatgcaa ttaagtgaag gtgaagaaag tagagctttt agttttgatg ttggtggtag
     5701 aggttatgtt ttaagagtta atagttgtgc tgatggtttt tataaagata gatatgttta
     5761 tagacatttt gctagtgctg ctttacctat tcctgaagtt ttagatattg gtgaatttag
     5821 tgaaagttta acttattgta ttagtagaag agcacaaggt gttactttac aagatttacc
     5881 tgaaactgaa ttacctgctg ttttacaacc tgttgctgaa gctatggatg ctattgcggc
     5941 ggctgatctc agtcaaactt cgggttttgg tccttttggt cctcaaggta ttggtcaata
     6001 tactacttgg agagatttta tttgtgctat tgctgatcct catgtttatc attggcaaac
     6061 tgttatggat gatactgtta gtgctagtgt tgctcaagca ttagatgaat taatgttatg
     6121 ggctgaagat tgtcctgaag ttagacattt agttcatgct gattttggta gtaataatgt
     6181 tttaactgat aatggtagaa ttactgctgt tattgattgg agtgaagcta tgtttggtga
     6241 tagtcaatat gaagttgcta atattttttt ttggagacct tggttagctt gtatggaaca
     6301 acaaactaga tattttgaaa gaagacatcc tgaattagct ggtagtccta gattaagagc
     6361 ttatatgtta agaattggtt tagatcaatt atatcaaagt ttagttgatg gtaattttga
     6421 tgatgctgct tgggctcaag gtagatgtga tgctattgtt agaagtggtg ctggtactgt
     6481 tggtagaact caaattgcta gaagaagtgc tgctgtttgg actgatggtt gtgttgaagt
     6541 tttagctgat agtggtaata gaagacctag tactagacct agagctaaag aataacaatt
     6601 cgttcaagcc gagatcggct tacgcggtta atcttcaata acatgtaaac cacgacgttt
     6661 taaatcttct aatgaatcta acattcctct tattaattca acaggatgtc cttcccaatc
     6721 ttcaacaaca ccaacaattc ttaaaggttc acaagttcta taagattgtg ttggattacc
     6781 aggaaatttt ttatttgtaa gatttggatc atcttcaaat ggtcctgttg gttcaactat
     6841 atatatataa ccacgacctt caagaccaga tagtgacata gcaagttcag ctccccaaac
     6901 tgctggttcc attaaagctg aaaaataaat atgtttaaga attcgaccat cttcaaaatg
     6961 agaaataaat cctgtagtta gtaaatcacc aattgctaaa ttagctttag ttccatgata
     7021 aaatggtcct tgtacttgtt tataattatc atgagaaata ggaatccaat cttttaccat
     7081 gacgttacct cctcatgatg tttaactttg ttttagggcg actgccctgc tgcgtaacat
     7141 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     7201 cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgcgg ccgtacccga
     7261 gcttcaagga agatttccta ttaaggttga acttaagagc ttaagcatag acgatttgaa
     7321 aaaaatttta aaacaaacaa aaaattcttt aataaaacaa tatgttgcga tgtttaaggt
     7381 ttatgattta gatttaaagt ttagtgagga agctatagat agaattgcag agcttacttt
     7441 taatatgaat cttgagagtg aaaatcttgg tgccagaaga cttcacggtg ttatggaaat
     7501 agtgcttgca gatctttttt ttgaagtgcc tggcagtaag ttgaaaaaat ttgaaataaa
     7561 cttggactat gttaataaaa aaatacaaat taacgaacaa aaagatttga attattatat
     7621 aatttagtta aaagcaattt taaatgaggg aggtttccat catatggata gaagtggtaa
     7681 acctgaatta actgctacta gtgttgaaaa atttttaatt gaaaaatttg atagtgttag
     7741 tgatttaatg caattaagtg aaggtgaaga aagtagagct tttagttttg atgttggtgg
     7801 tagaggttat gttttaagag ttaatagttg tgctgatggt ttttataaag atagatatgt
     7861 ttatagacat tttgctagtg ctgctttacc tattcctgaa gttttagata ttggtgaatt
     7921 tagtgaaagt ttaacttatt gtattagtag aagagcacaa ggtgttactt tacaagattt
     7981 acctgaaact gaattacctg ctgttttaca acctgttgct gaagctatgg atgctattgc
     8041 ggcggctgat ctcagtcaaa cttcgggttt tggtcctttt ggtcctcaag gtattggtca
     8101 atatactact tggagagatt ttatttgtgc tattgctgat cctcatgttt atcattggca
     8161 aactgttatg gatgatactg ttagtgctag tgttgctcaa gcattagatg aattaatgtt
     8221 atgggctgaa gattgtcctg aagttagaca tttagttcat gctgattttg gtagtaataa
     8281 tgttttaact gataatggta gaattactgc tgttattgat tggagtgaag ctatgtttgg
     8341 tgatagtcaa tatgaagttg ctaatatttt tttttggaga ccttggttag cttgtatgga
     8401 acaacaaact agatattttg aaagaagaca tcctgaatta gctggtagtc ctagattaag
     8461 agcttatatg ttaagaattg gtttagatca attatatcaa agtttagttg atggtaattt
     8521 tgatgatgct gcttgggctc aaggtagatg tgatgctatt gttagaagtg gtgctggtac
     8581 tgttggtaga actcaaattg ctagaagaag tgctgctgtt tggactgatg gttgtgttga
     8641 agttttagct gatagtggta atagaagacc tagtactaga cctagagcta aagaataaca
     8701 attcgttcaa gccgagatcg gcttacgcgt
//
