LOCUS       sequence-227345-        6675 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Kanamycin-resistant E.coli/B. burgdorferi shuttle vector; Encodes 
            bb0526 promoter driven mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6675)
  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 6675)
  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. 2018 Oct 12. pii: AEM.01824-18. doi: 
            10.1128/AEM.01824-18."
COMMENT     Sequence Label: pBSV2_P0526-mCherryBb
FEATURES             Location/Qualifiers
     source          1..6675
                     /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(1014..1031)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1014..1030)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1023..1045)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1597..2406
                     /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(1664..1683)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2575..3163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3064..3083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6640..6661
                     /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 ttcgagctcc ttgttttcaa tgataggttt
      121 tttaggtaga gacttgtaag ggtttgttcc tggaatataa tttttaggaa ttttcatata
      181 ttttactcct atttaactag ctaacctata attctattat tttttttgtt tttatgcaat
      241 caatattgat attttgattt tttatgatta gaatcatcat ggatccagga ggttcatggt
      301 tagtaaaggt gaagaagata atatggctat tattaaagaa tttatgagat ttaaagttca
      361 catggaaggt agtgttaatg gtcatgaatt tgaaattgaa ggtgaaggtg aaggtagacc
      421 ttatgaaggt actcaaactg ctaaattaaa agttactaaa ggtggtcctt taccttttgc
      481 ttgggatatt ttaagtcctc aatttatgta tggtagtaaa gcatacgtta aacatcctgc
      541 tgatattcct gattatttaa aattaagttt tcctgaaggt tttaaatggg aaagagttat
      601 gaattttgaa gatggtggtg ttgttactgt tactcaagat agtagtttac aagatggtga
      661 atttatttat aaagttaaat taagaggtac taattttcct agtgatggtc ctgttatgca
      721 aaaaaaaact atgggttggg aagctagtag tgaaagaatg tatcctgaag atggtgcttt
      781 aaaaggtgaa attaaacaaa gattaaaatt aaaagatggt ggtcattatg atgctgaagt
      841 taaaactact tataaagcta aaaaacctgt tcaattacct ggtgcttata atgttaatat
      901 taaattagat attacttcgc ataatgaaga ttatactatt gttgaacaat atgaaagagc
      961 tgaaggtaga catagtacag gtggtatgga tgaattatat aaataagctt ggcactggcc
     1021 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1081 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1141 caacagttgc gcagcctgaa tggcgaatgg cgcggccgcc ctaggttaat acccgagctt
     1201 caaggaagat ttcctattaa ggttgaactt aagagcttaa gcatagacga tttgaaaaaa
     1261 attttaaaac aaacaaaaaa ttctttaata aaacaatatg ttgcgatgtt taaggtttat
     1321 gatttagatt taaagtttag tgaggaagct atagatagaa ttgcagagct tacttttaat
     1381 atgaatcttg agagtgaaaa tcttggtgcc agaagacttc acggtgttat ggaaatagtg
     1441 cttgcagatc ttttttttga agtgcctggc agtaagttga aaaaatttga aataaacttg
     1501 gactatgtta ataaaaaaat acaaattaac gaacaaaaag atttgaatta ttatataatt
     1561 tagttaaaag caattttaaa tgagggaggt ttccatatgg gccatattca acgggaaacg
     1621 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1681 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1741 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1801 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1861 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1921 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1981 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2041 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2101 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2161 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2221 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2281 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2341 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2401 ttctaatcag actcgagtgg ccgaggagca ggactgacac gtgctaaaac ttcattttta
     2461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2641 ggtttgtttg ccggatcaag agctaccaac tcttttttcg aaggtaactg gcttcagcag
     2701 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcactaga
     3241 tccccggcaa gttcatcctt agggttttca tatctcttaa aaagtcaatt tctgcttgat
     3301 taatttcttg tggattattg tttttgcggt ttttcatttt ttatctcctt gagtattgaa
     3361 ttttttctta tatataaata tatagcaaaa actatttttg ccaacttttt tacaaaaaat
     3421 tttttattaa aacacttagg gctttgctaa attctctttt aaaagaactt agtaaagccc
     3481 tatggattta agaactgatt atacttacgt agtaaaatac tacagatatt aatagtatag
     3541 accatatcaa aatcataatc aatttatatt tatctaaaat tgtagtccat acaaaataca
     3601 aaaaaattag gtttttctaa attctatttt aaaagaactt agtaacacct aatctctaaa
     3661 ttacttacaa aattctaaaa ttagattttt agttcttcat atttctttaa aagtttttta
     3721 agaaaatctt tttgattttc aaaaatttca tgcatcataa aactagtaaa tttggatttt
     3781 tttttgtaaa aatcataact ttcttgggtt ttaagttgaa atcttaatgg ttttattgga
     3841 ttttgcctag atttttttaa cgccggactt tctttatctt ttataaaatt ataagaattt
     3901 ttaataccat tttctattaa ataatcttct tttataagcc cctcttgcag tgcagttgct
     3961 aatcttaaat acttatacgc ttgagttttt gctacatcaa agtcttttat aaaatcaaga
     4021 aaagttttat aaccatcaaa tttataatat tgattatctt ttatttcttt taaaatttta
     4081 atattttcca ccctgtggaa tatttctttt ttgaaattgt catttaactt ttcctttagt
     4141 gtcttgtaat aagaaatttt actttcatta atcattaaat tttgttcaac ataagtatct
     4201 atcttttttg ttaaaacttt aacaacacgt tttttttcca taaaaatttt cctttatttt
     4261 tgcaacattt tccttaattg atataattgt tactatttta gattttttaa tatccactta
     4321 agaattaatc cctcactaga atatttttta ttatagaatt catatacatg tttttctaaa
     4381 tctataaatt tatcaacaat tgctttatta tagggtgtat ctatcttttc ttttcttaat
     4441 aatcgtctcg tattctttaa ataacaacaa acactgccgg ttttaaacct aaattctatg
     4501 taatatattt ttgaaaaagt atatacattt tcaaccccgt ttatttcata ctttacaaaa
     4561 acattattta ttggttttct ataaccatag taaatgccta aaaatttgtc cccgtctttc
     4621 ataggatata gataaagttt ttgaatctct gatttatcta ataatttttc aaaaaaaatt
     4681 atacaccgct tttgtttgtt actaactact ctatgcaatg cctttaatat ttttgtgtga
     4741 tatatatttt tatcaccttt tttttctttc agcaggaaga tactttcttt tggaaattta
     4801 attttttgtt catttattct tgcagcctta agtccgattt tcatatttaa tccttatata
     4861 gcctttctat ttaattcttt taaactcaat aaactttttt ggtttttcat tagttctaat
     4921 aattcgcaat aatatgtgca aaacacttta ttgtattcta ttttcttttg gttattcaaa
     4981 tattctttta taattggctt tagaagttca atatttactt ctttttttag ttgttcaata
     5041 aggatattga aaatgtttgt ttttaaatct tgataatttt gttgagaatt ttcttttttt
     5101 ctttcaattg acttctctaa tttacgcttt atattgttta aatcgctata tttatgactt
     5161 tcaataataa aatgaggctt aaatttgtaa ttttcgtata ttttttggaa acttgtttct
     5221 aattgtttag aattatattc atttttttca aattcttttt gagtgttgca tagaatttcc
     5281 tttaattttt cttgtttttc tttcatacaa gatttagtaa aatgtatatt tttatttttt
     5341 agtaatttaa tttctattat ttttatgatt ttaagtattt tgattatatt atctttatca
     5401 atatttaaat tgaataaaat tgtaagaaaa attgaaagaa tttttttaca taaaaagtta
     5461 catttattga agtaatttct tacttgatac ttttctattt cattaatctt tctttcttct
     5521 tttatattat ttttattatt taaacactcc tttaaatcta cactcccatt tttagtaaat
     5581 ctatctttaa ggccccttga aactctattt tgaaatctta aatctttttt ttcttgaaag
     5641 tatttataga ttttttggta acattctttt tttgaaaaat taagttgata gtatatttcg
     5701 gtccccaaat ttacccctaa gtgtttgtga taattagttg tgactttaaa ttctttttct
     5761 aatttgtaaa ggtaattttg cagtgttttt agtttaacgg gtgtttgact atttctttta
     5821 aggttttcat taaagtaata gagtatgttt ttttgtgtgt atttcttata tgttttattt
     5881 atgtacgcta gtgttgagat aaggactact aatttatact gtaatttatt gtagcaagtt
     5941 gtggttttac tggttttgtt tatattagat aagttttcca tttatagctc cttgaatacc
     6001 ggatttttta tctataaagt atatatatca caattttgat ttaaaagtaa attctttttt
     6061 taaaaataca aaaatttttt ataaattata tagaccatga tattgtatta atactaattg
     6121 attttaattt aaggttgaac taaaataaac atagttttgt agaaaataat ttttcattat
     6181 tttgctactt gatattggat cgtaaaaata ttagggcttt acaaagttct tttaaaagag
     6241 aatttagtaa agccctaagt cttttaataa aaaatttttt gtaaaaaagt tggcaaaaat
     6301 agtttttgct atatatttat atataagaaa aaattcaata ctcaaggaga taaaaaatgg
     6361 aaaacttaaa caataatcca caaaaaatta atcaaataaa taagcaaact aattttattg
     6421 gttttgaaaa ttttatagaa cttgataatc aagttaatca aactaaagaa aaaagtaaat
     6481 taagtaagat aggtaagaaa ttacccggaa taagcagtca agtctagtgc ggccgcgccc
     6541 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     6601 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     6661 ttaggcaccc caggc
//
