LOCUS       sequence-223338-        6134 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 6134)
  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 6134)
  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..6134
                     /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(4360..4377)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4360..4376)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4369..4391)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4943..5734
                     /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"
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 tatgaccatg
     4261 attacgaatt cgagctcggt acaccggtct cgaggacgtc gctagcggat cccggggtac
     4321 cgatcctcta gagtcgacct gcaggcatgc aagcttggca ctggccgtcg ttttacaacg
     4381 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4441 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4501 cctgaatggc gaatggcgcg gccgccctag gttaataccc gagcttcaag gaagatttcc
     4561 tattaaggtt gaacttaaga gcttaagcat agacgatttg aaaaaaattt taaaacaaac
     4621 aaaaaattct ttaataaaac aatatgttgc gatgtttaag gtttatgatt tagatttaaa
     4681 gtttagtgag gaagctatag atagaattgc agagcttact tttaatatga atcttgagag
     4741 tgaaaatctt ggtgccagaa gacttcacgg tgttatggaa atagtgcttg cagatctttt
     4801 ttttgaagtg cctggcagta agttgaaaaa atttgaaata aacttggact atgttaataa
     4861 aaaaatacaa attaacgaac aaaaagattt gaattattat ataatttagt taaaagcaat
     4921 tttaaatgag ggaggtttcc atatgaggga agcggtgatc gccgaagtat cgactcaact
     4981 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     5041 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     5101 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     5161 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     5221 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     5281 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     5341 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     5401 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     5461 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     5521 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     5581 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     5641 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     5701 gaaaggcgag atcaccaagg tagtcggcaa ataagacgtc gccggagcgg tcgagttctg
     5761 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     5821 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     5881 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     5941 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     6001 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     6061 ggactgacac gtgctaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6121 tgataatctc atga
//
