LOCUS       sequence-227367-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kanamycin-resistant E.coli/B. burgdorferi shuttle vector; Encodes 
            bb0031 promoter driven mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  AUTHORS   Takacs CN, Kloos ZA, Scott M, Rosa PA, Jacobs-Wagner C
  TITLE     Fluorescent Proteins, Promoters, and Selectable Markers for 
            Applications in the Lyme Disease Spirochete Borrelia burgdorferi
  JOURNAL   Appl Environ Microbiol. 2018 Oct 12. pii: AEM.01824-18. doi: 
            10.1128/AEM.01824-18.
  PUBMED    30315081
REFERENCE   2  (bases 1 to 6692)
  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. 2018 Oct 12. pii: AEM.01824-18. doi: 
            10.1128/AEM.01824-18."
COMMENT     Sequence Label: pBSV2_P0031-mCherryBb
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1031..1048)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1031..1047)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1040..1062)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1614..2423
                     /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="MGHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1681..1700)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2592..3180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3081..3100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6657..6678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgaca tgattacgaa ttcgagctcg ttgatattaa acttaaaagc
      121 aatattattg ttgtaagaaa tacgtaaacc ttaggctcta ttcctaggat tggctttaat
      181 ttaacttcaa aaagctcaac atttgaattc tcgtcaacct ctttcatctt ttaaagtata
      241 gaaaagtatt taaataagtg tcaatatttt gtattattta attcttgagg ttaggttgga
      301 tccaggaggt tcatggttag taaaggtgaa gaagataata tggctattat taaagaattt
      361 atgagattta aagttcacat ggaaggtagt gttaatggtc atgaatttga aattgaaggt
      421 gaaggtgaag gtagacctta tgaaggtact caaactgcta aattaaaagt tactaaaggt
      481 ggtcctttac cttttgcttg ggatatttta agtcctcaat ttatgtatgg tagtaaagca
      541 tacgttaaac atcctgctga tattcctgat tatttaaaat taagttttcc tgaaggtttt
      601 aaatgggaaa gagttatgaa ttttgaagat ggtggtgttg ttactgttac tcaagatagt
      661 agtttacaag atggtgaatt tatttataaa gttaaattaa gaggtactaa ttttcctagt
      721 gatggtcctg ttatgcaaaa aaaaactatg ggttgggaag ctagtagtga aagaatgtat
      781 cctgaagatg gtgctttaaa aggtgaaatt aaacaaagat taaaattaaa agatggtggt
      841 cattatgatg ctgaagttaa aactacttat aaagctaaaa aacctgttca attacctggt
      901 gcttataatg ttaatattaa attagatatt acttcgcata atgaagatta tactattgtt
      961 gaacaatatg aaagagctga aggtagacat agtacaggtg gtatggatga attatataaa
     1021 taagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1081 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1141 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ggccgcccta
     1201 ggttaatacc cgagcttcaa ggaagatttc ctattaaggt tgaacttaag agcttaagca
     1261 tagacgattt gaaaaaaatt ttaaaacaaa caaaaaattc tttaataaaa caatatgttg
     1321 cgatgtttaa ggtttatgat ttagatttaa agtttagtga ggaagctata gatagaattg
     1381 cagagcttac ttttaatatg aatcttgaga gtgaaaatct tggtgccaga agacttcacg
     1441 gtgttatgga aatagtgctt gcagatcttt tttttgaagt gcctggcagt aagttgaaaa
     1501 aatttgaaat aaacttggac tatgttaata aaaaaataca aattaacgaa caaaaagatt
     1561 tgaattatta tataatttag ttaaaagcaa ttttaaatga gggaggtttc catatgggcc
     1621 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     1681 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     1741 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1801 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1861 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     1921 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1981 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2041 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2101 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     2161 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2221 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2281 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2341 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2401 tgatgctcga tgagtttttc taatcagact cgagtggccg aggagcagga ctgacacgtg
     2461 ctaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2521 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2581 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2641 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct tttttcgaag
     2701 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2761 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2821 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2881 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2941 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3001 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3061 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3121 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3181 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3241 ttctttcctg cactagatcc ccggcaagtt catccttagg gttttcatat ctcttaaaaa
     3301 gtcaatttct gcttgattaa tttcttgtgg attattgttt ttgcggtttt tcatttttta
     3361 tctccttgag tattgaattt tttcttatat ataaatatat agcaaaaact atttttgcca
     3421 acttttttac aaaaaatttt ttattaaaac acttagggct ttgctaaatt ctcttttaaa
     3481 agaacttagt aaagccctat ggatttaaga actgattata cttacgtagt aaaatactac
     3541 agatattaat agtatagacc atatcaaaat cataatcaat ttatatttat ctaaaattgt
     3601 agtccataca aaatacaaaa aaattaggtt tttctaaatt ctattttaaa agaacttagt
     3661 aacacctaat ctctaaatta cttacaaaat tctaaaatta gatttttagt tcttcatatt
     3721 tctttaaaag ttttttaaga aaatcttttt gattttcaaa aatttcatgc atcataaaac
     3781 tagtaaattt ggattttttt ttgtaaaaat cataactttc ttgggtttta agttgaaatc
     3841 ttaatggttt tattggattt tgcctagatt tttttaacgc cggactttct ttatctttta
     3901 taaaattata agaattttta ataccatttt ctattaaata atcttctttt ataagcccct
     3961 cttgcagtgc agttgctaat cttaaatact tatacgcttg agtttttgct acatcaaagt
     4021 cttttataaa atcaagaaaa gttttataac catcaaattt ataatattga ttatctttta
     4081 tttcttttaa aattttaata ttttccaccc tgtggaatat ttcttttttg aaattgtcat
     4141 ttaacttttc ctttagtgtc ttgtaataag aaattttact ttcattaatc attaaatttt
     4201 gttcaacata agtatctatc ttttttgtta aaactttaac aacacgtttt ttttccataa
     4261 aaattttcct ttatttttgc aacattttcc ttaattgata taattgttac tattttagat
     4321 tttttaatat ccacttaaga attaatccct cactagaata ttttttatta tagaattcat
     4381 atacatgttt ttctaaatct ataaatttat caacaattgc tttattatag ggtgtatcta
     4441 tcttttcttt tcttaataat cgtctcgtat tctttaaata acaacaaaca ctgccggttt
     4501 taaacctaaa ttctatgtaa tatatttttg aaaaagtata tacattttca accccgttta
     4561 tttcatactt tacaaaaaca ttatttattg gttttctata accatagtaa atgcctaaaa
     4621 atttgtcccc gtctttcata ggatatagat aaagtttttg aatctctgat ttatctaata
     4681 atttttcaaa aaaaattata caccgctttt gtttgttact aactactcta tgcaatgcct
     4741 ttaatatttt tgtgtgatat atatttttat cacctttttt ttctttcagc aggaagatac
     4801 tttcttttgg aaatttaatt ttttgttcat ttattcttgc agccttaagt ccgattttca
     4861 tatttaatcc ttatatagcc tttctattta attcttttaa actcaataaa cttttttggt
     4921 ttttcattag ttctaataat tcgcaataat atgtgcaaaa cactttattg tattctattt
     4981 tcttttggtt attcaaatat tcttttataa ttggctttag aagttcaata tttacttctt
     5041 tttttagttg ttcaataagg atattgaaaa tgtttgtttt taaatcttga taattttgtt
     5101 gagaattttc tttttttctt tcaattgact tctctaattt acgctttata ttgtttaaat
     5161 cgctatattt atgactttca ataataaaat gaggcttaaa tttgtaattt tcgtatattt
     5221 tttggaaact tgtttctaat tgtttagaat tatattcatt tttttcaaat tctttttgag
     5281 tgttgcatag aatttccttt aatttttctt gtttttcttt catacaagat ttagtaaaat
     5341 gtatattttt attttttagt aatttaattt ctattatttt tatgatttta agtattttga
     5401 ttatattatc tttatcaata tttaaattga ataaaattgt aagaaaaatt gaaagaattt
     5461 ttttacataa aaagttacat ttattgaagt aatttcttac ttgatacttt tctatttcat
     5521 taatctttct ttcttctttt atattatttt tattatttaa acactccttt aaatctacac
     5581 tcccattttt agtaaatcta tctttaaggc cccttgaaac tctattttga aatcttaaat
     5641 cttttttttc ttgaaagtat ttatagattt tttggtaaca ttcttttttt gaaaaattaa
     5701 gttgatagta tatttcggtc cccaaattta cccctaagtg tttgtgataa ttagttgtga
     5761 ctttaaattc tttttctaat ttgtaaaggt aattttgcag tgtttttagt ttaacgggtg
     5821 tttgactatt tcttttaagg ttttcattaa agtaatagag tatgtttttt tgtgtgtatt
     5881 tcttatatgt tttatttatg tacgctagtg ttgagataag gactactaat ttatactgta
     5941 atttattgta gcaagttgtg gttttactgg ttttgtttat attagataag ttttccattt
     6001 atagctcctt gaataccgga ttttttatct ataaagtata tatatcacaa ttttgattta
     6061 aaagtaaatt ctttttttaa aaatacaaaa attttttata aattatatag accatgatat
     6121 tgtattaata ctaattgatt ttaatttaag gttgaactaa aataaacata gttttgtaga
     6181 aaataatttt tcattatttt gctacttgat attggatcgt aaaaatatta gggctttaca
     6241 aagttctttt aaaagagaat ttagtaaagc cctaagtctt ttaataaaaa attttttgta
     6301 aaaaagttgg caaaaatagt ttttgctata tatttatata taagaaaaaa ttcaatactc
     6361 aaggagataa aaaatggaaa acttaaacaa taatccacaa aaaattaatc aaataaataa
     6421 gcaaactaat tttattggtt ttgaaaattt tatagaactt gataatcaag ttaatcaaac
     6481 taaagaaaaa agtaaattaa gtaagatagg taagaaatta cccggaataa gcagtcaagt
     6541 ctagtgcggc cgcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6601 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6661 gtgagttagc tcactcatta ggcaccccag gc
//
