LOCUS       sequence-227360-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty E. coli - B. burgdorferi shuttle vector; streptomycin 
            resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  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 6129)
  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: pKFSS1_2
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..44)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(27..43)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(36..58)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             610..1401
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1873..2461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2362..2381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5939..5960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5975..5980,5981..5998,5999..6005)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6013..6029
                     /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     6018..6040
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6037..6053
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6037..6053
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tcgacctgca ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
       61 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      121 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      181 tggcgcggcc gccctaggtt aatacccgag cttcaaggaa gatttcctat taaggttgaa
      241 cttaagagct taagcataga cgatttgaaa aaaattttaa aacaaacaaa aaattcttta
      301 ataaaacaat atgttgcgat gtttaaggtt tatgatttag atttaaagtt tagtgaggaa
      361 gctatagata gaattgcaga gcttactttt aatatgaatc ttgagagtga aaatcttggt
      421 gccagaagac ttcacggtgt tatggaaata gtgcttgcag atcttttttt tgaagtgcct
      481 ggcagtaagt tgaaaaaatt tgaaataaac ttggactatg ttaataaaaa aatacaaatt
      541 aacgaacaaa aagatttgaa ttattatata atttagttaa aagcaatttt aaatgaggga
      601 ggtttccata tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
      661 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
      721 gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg
      781 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
      841 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
      901 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
      961 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     1021 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     1081 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     1141 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
     1201 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
     1261 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
     1321 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc
     1381 accaaggtag tcggcaaata atgacgtcgc cggagcggtc gagttctgga ccgaccggct
     1441 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     1501 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     1561 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
     1621 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
     1681 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
     1741 gctaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1801 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1861 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1921 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1981 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2041 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2101 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2161 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2221 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2281 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2341 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2401 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2461 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2521 gttctttcct gcactagatc cccggcaagt tcatccttag ggttttcata tctcttaaaa
     2581 agtcaatttc tgcttgatta atttcttgtg gattattgtt tttgcggttt ttcatttttt
     2641 atctccttga gtattgaatt ttttcttata tataaatata tagcaaaaac tatttttgcc
     2701 aactttttta caaaaaattt tttattaaaa cacttagggc tttgctaaat tctcttttaa
     2761 aagaacttag taaagcccta tggatttaag aactgattat acttacgtag taaaaatact
     2821 acagatatta atagtataga ccatatcaaa atcataatca atttatattt atctaaaatt
     2881 gtagtccata caaaatacaa aaaaattagg tttttctaaa ttctatttta aaagaactta
     2941 gtaacaccta atctctaaat tacttacaaa attctaaaat tagattttta gttcttcata
     3001 tttctttaaa agttttttaa gaaaatcttt ttgattttca aaaatttcat gcatcataaa
     3061 actagtaaat ttggattttt ttttgtaaaa atcataactt tcttgggttt taagttgaaa
     3121 tcttaatggt tttattggat tttgcctaga tttttttaac gccggacttt ctttatcttt
     3181 tataaaatta taagaatttt taataccatt ttctattaaa taatcttctt ttataagccc
     3241 ctcttgcagt gcagttgcta atcttaaata cttatacgct tgagtttttg ctacatcaaa
     3301 gtcttttata aaatcaagaa aagttttata accatcaaat ttataatatt gattatcttt
     3361 tatttctttt aaaattttaa tattttccac cctgtggaat atttcttttt tgaaattgtc
     3421 atttaacttt tcctttagtg tcttgtaata agaaatttta ctttcattaa tcattaaatt
     3481 ttgttcaaca taagtatcta tcttttttgt taaaacttta acaacacgtt ttttttccat
     3541 aaaaattttc ctttattttt gcaacatttt ccttaattga tataattgtt actattttag
     3601 attttttaat atccacttaa gaattaatcc ctcactagaa tattttttat tatagaattc
     3661 atatacatgt ttttctaaat ctataaattt atcaacaatt gctttattat agggtgtatc
     3721 tatcttttct tttcttaata atcgtctcat attctttaaa taacaacaaa cactgccggt
     3781 tttaaaccta aattctatgt aatatatttt tgaaaaagta tatacatttt caaccccgtt
     3841 tatttcatac tttacaaaaa cattatttat tggttttcta taaccatagt aaatgcctaa
     3901 aaatttgtcc ccgtctttca taggatatag ataaagtttt tgaatctctg atttatctaa
     3961 taatttttca aaaaaaatta tacaccgctt ttgtttgtta ctaactactc tatgcaatgc
     4021 ctttaatatt tttgtgtgat atatattttt atcacctttt ttttctttca gcaggaagat
     4081 actttctttt ggaaatttaa ttttttgttc atttattctt gcagccttaa gtccgatttt
     4141 catatttaat ccttatatag cctttctatt taattctttt aaactcaata aacttttttg
     4201 gtttttcatt agttctaata attcgcaata atatgtgcaa aacactttat tgtattctat
     4261 tttcttttgg ttattcaaat attcttttat aattggcttt agaagttcaa tatttacttc
     4321 tttttttagt tgttcaataa ggatattgaa aatgtttgtt tttaaatctt gataattttg
     4381 ttgagaattt tctttttttc tttcaattga cttctctaat ttacgcttta tattgtttaa
     4441 atcgctatat ttatgacttt caataataaa atgaggctta aatttgtaat tttcgtatat
     4501 tttttggaaa cttgtttcta attgtttaga attatattca tttttttcaa attctttttg
     4561 agtgttgcat agaatttcct ttaatttttc ttgtttttct ttcatacaag atttagtaaa
     4621 atgtatattt ttatttttta gtaatttaat ttctattatt tttatgattt taagtatttt
     4681 gattatatta tctttatcaa tatttaaatt gaataaaatt gtaagaaaaa ttgaaagaat
     4741 ttttttacat aaaaagttac atttattgaa gtaatttctt acttgatact tttctatttc
     4801 attaatcttt ctttcttctt ttatattatt tttattattt aaacactcct ttaaatctac
     4861 actcccattt ttagtaaatc tatctttaag gccccttgaa actctatttt gaaatcttaa
     4921 atcttttttt tcttgaaagt atttatagat tttttggtaa cattcttttt ttgaaaaatt
     4981 aagttgatag tatatttcgg tccccaaatt tacccctaag tgtttgtgat aattagttgt
     5041 gactttaaat tctttttcta atttgtaaag gtaattttgc agtgttttta gtttaacggg
     5101 tgtttgacta tttcttttaa ggttttcatt aaagtaatag agtatgtttt tttgtgtgta
     5161 tttcttatat gttttattta tgtacgctag tgttgagata aggactacta atttatactg
     5221 taatttattg tagcaagttg tggttttact ggttttgttt atattagata agttttccat
     5281 ttatagctcc ttgaataccg gattttttat ctataaagta tatatatcac aattttgatt
     5341 taaaagtaaa ttcttttttt aaaaatacaa aaatttttta taaattatat agaccatgat
     5401 attgtattaa tactaattga ttttaattta aggttgaact aaaataaaca tagttttgta
     5461 gaaaataatt tttcattatt ttgctacttg atattggatc gtaaaaatat tagggcttta
     5521 caaagttctt ttaaaagaga atttagtaaa gccctaagtc ttttaataaa aaattttttg
     5581 taaaaaagtt ggcaaaaata gtttttgcta tatatttata tataagaaaa aattcaatac
     5641 tcaaggagat aaaaaatgga aaacttaaac aataatccac aaaaaattaa tcaaataaat
     5701 aagcaaacta attttattgg ttttgaaaat tttatagaac ttgataatca agttaatcaa
     5761 actaaagaaa aaagtaaatt aagtaagata ggtaagaaat tacccggaat aagcagtcaa
     5821 gtctagtgcg gccgcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5881 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5941 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6001 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta
     6061 cgaattcgag ctcggtacac cggtctcgag gacgtcgcta gcggatcccg gggtaccgat
     6121 cctctagag
//
