LOCUS       sequence-223351-        6698 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 6698)
  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 6698)
  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..6698
                     /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"
     primer_bind     complement(5208..5225)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5208..5224)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5217..5239)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5791..6600
                     /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(5858..5877)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
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 tatgacatga
     4261 ttacgaattc gagctcgttg atattaaact taaaagcaat attattgttg taagaaatac
     4321 gtaaacctta ggctctattc ctaggattgg ctttaattta acttcaaaaa gctcaacatt
     4381 tgaattctcg tcaacctctt tcatctttta aagtatagaa aagtatttaa ataagtgtca
     4441 atattttgta ttatttaatt cttgaggtta ggttggatcc aggaggttca tggttagtaa
     4501 aggtgaagaa gataatatgg ctattattaa agaatttatg agatttaaag ttcacatgga
     4561 aggtagtgtt aatggtcatg aatttgaaat tgaaggtgaa ggtgaaggta gaccttatga
     4621 aggtactcaa actgctaaat taaaagttac taaaggtggt cctttacctt ttgcttggga
     4681 tattttaagt cctcaattta tgtatggtag taaagcatac gttaaacatc ctgctgatat
     4741 tcctgattat ttaaaattaa gttttcctga aggttttaaa tgggaaagag ttatgaattt
     4801 tgaagatggt ggtgttgtta ctgttactca agatagtagt ttacaagatg gtgaatttat
     4861 ttataaagtt aaattaagag gtactaattt tcctagtgat ggtcctgtta tgcaaaaaaa
     4921 aactatgggt tgggaagcta gtagtgaaag aatgtatcct gaagatggtg ctttaaaagg
     4981 tgaaattaaa caaagattaa aattaaaaga tggtggtcat tatgatgctg aagttaaaac
     5041 tacttataaa gctaaaaaac ctgttcaatt acctggtgct tataatgtta atattaaatt
     5101 agatattact tcgcataatg aagattatac tattgttgaa caatatgaaa gagctgaagg
     5161 tagacatagt acaggtggta tggatgaatt atataaataa gcttggcact ggccgtcgtt
     5221 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5281 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5341 ttgcgcagcc tgaatggcga atggcgcggc cgccctaggt taatacccga gcttcaagga
     5401 agatttccta ttaaggttga acttaagagc ttaagcatag acgatttgaa aaaaatttta
     5461 aaacaaacaa aaaattcttt aataaaacaa tatgttgcga tgtttaaggt ttatgattta
     5521 gatttaaagt ttagtgagga agctatagat agaattgcag agcttacttt taatatgaat
     5581 cttgagagtg aaaatcttgg tgccagaaga cttcacggtg ttatggaaat agtgcttgca
     5641 gatctttttt ttgaagtgcc tggcagtaag ttgaaaaaat ttgaaataaa cttggactat
     5701 gttaataaaa aaatacaaat taacgaacaa aaagatttga attattatat aatttagtta
     5761 aaagcaattt taaatgaggg aggtttccat atgggccata ttcaacggga aacgtcgagg
     5821 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     5881 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     5941 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     6001 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     6061 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     6121 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     6181 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     6241 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     6301 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     6361 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     6421 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     6481 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     6541 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     6601 tcagactcga gtggccgagg agcaggactg acacgtgcta aaacttcatt tttaatttaa
     6661 aaggatctag gtgaagatcc tttttgataa tctcatga
//
