LOCUS       sequence-228482-        6500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gentamicin-resistant E.coli/B. burgdorferi shuttle vector; Encodes 
            flagellin promoter driven msfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  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 6500)
  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: pBSV2G_PflaB-msfGFPBb
FEATURES             Location/Qualifiers
     source          1..6500
                     /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(1181..1198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1181..1197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1190..1212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1764..2297
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      2400..2988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2889..2908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6465..6486
                     /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 ttcgagctct gtctgtcgcc tcttgtggct
      121 tccggaggag ttatttataa taaaataaag aattacgact tgaaaatttt taatcaaaaa
      181 aataaagttg aagattataa aaagaagtac gaagatagag agagaaaagt ggaaggtgaa
      241 cttaatacct tggattttac cgttaagcgc atgaaagatc aagaaaatac attaaaggct
      301 tttgatttta atcaaagaaa taaataataa taattatttt taatgctatt gctatttgcg
      361 tttctttttt tttaattttt gtgctattct ttttaacagg caaaaggatt tgccaaagtc
      421 agaaatttaa attttatcat ggaggaatga ggatccatga gtaaaggtga agaattattt
      481 actggtgttg ttcctatttt agttgaatta gatggtgatg ttaatggtca taaatttagt
      541 gttagaggtg aaggtgaagg tgatgctact aatggtaaat taactttaaa atttatttgt
      601 actactggta aattacctgt tccttggcct actttagtta ctactttaac ttatggtgtt
      661 caatgtttta gtagatatcc tgatcatatg aaaagacatg atttttttaa aagtgctatg
      721 cctgaaggtt atgttcaaga aagaactatt agttttaaag atgatggtac ttataaaact
      781 agagctgaag ttaaatttga aggtgatact ttagttaata gaattgaatt aaaaggtatt
      841 gattttaaag aagatggtaa tattttaggt cataaattag aatataattt taatagtcat
      901 aatgtttata ttactgctga taaacaaaaa aatggtatta aagctaattt taaaattaga
      961 cataatgttg aagatggtag tgttcaatta gctgatcatt atcaacaaaa tactcctatt
     1021 ggtgatggtc ctgttttatt acctgataat cattatttaa gtactcaaag taaattaagt
     1081 aaagatccta atgaaaaaag agatcatatg gttttattag aatttgttac tgctgctggt
     1141 attactcatg gtatggatga attatataaa taagcttggc actggccgtc gttttacaac
     1201 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1261 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1321 gcctgaatgg cgaatggcgc ggccgcccta ggttaatacc cgagcttcaa ggaagatttc
     1381 ctattaaggt tgaacttaag agcttaagca tagacgattt gaaaaaaatt ttaaaacaaa
     1441 caaaaaattc tttaataaaa caatatgttg cgatgtttaa ggtttatgat ttagatttaa
     1501 agtttagtga ggaagctata gatagaattg cagagcttac ttttaatatg aatcttgaga
     1561 gtgaaaatct tggtgccaga agacttcacg gtgttatgga aatagtgctt gcagatcttt
     1621 tttttgaagt gcctggcagt aagttgaaaa aatttgaaat aaacttggac tatgttaata
     1681 aaaaaataca aattaacgaa caaaaagatt tgaattatta tataatttag ttaaaagcaa
     1741 ttttaaatga gggaggtttc catatgttac gcagcagcaa cgatgttacg cagcagggca
     1801 gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg
     1861 gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag
     1921 acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta
     1981 gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg
     2041 cttacgttct gcccaagttt gagcagccgc gtagtgagat ctatatctat gatctcgcag
     2101 tctccggcga gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg
     2161 aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg
     2221 cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc
     2281 caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttacgcgtc caaaatccct
     2341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2521 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgca
     3061 ctagatcccc ggcaagttca tccttagggt tttcatatct cttaaaaagt caatttctgc
     3121 ttgattaatt tcttgtggat tattgttttt gcggtttttc attttttatc tccttgagta
     3181 ttgaattttt tcttatatat aaaatatagc aaaaactatt tttgccaact tttttacaaa
     3241 aaatttttta ttaaaacact tagggctttg ctaaattctc ttttaaaaga acttagtaaa
     3301 gccctatgga tttaagaact gattatactt acgtagtaaa aatactacag atattaatag
     3361 tatagaccat atcaaaatca taatcaattt atatttatct aaaattgtgg tccatacaaa
     3421 atacaaaaaa attaggtttt tctaaattct attttaaaag aacttagtaa cacctaatct
     3481 ctaaattact tacaaaattc taaaattaga tttttagttc ttcatatttc tttaaaagtt
     3541 ttttaagaaa atctttttga ttttcaaaaa tttcatgcat cataaaacta gtaaatttgg
     3601 attttttttt gtaaaaatca taactttctt gggttttaag ttgaaatctt aatggtttta
     3661 ttggattttg cctagatttt tttaacgccg gactttcttt atcttttata aaattataag
     3721 aatttttaat accattttct attaaataat cttcttttat aagcccctct tgcagtgcag
     3781 ttgctaatct taaatactta tacgcttgag tttttgctac atcaaagtct tttataaaat
     3841 caagaaaagt tttataacca tcaaatttat aatattgatt atcttttatt tcttttaaaa
     3901 ttttaatatt ttccaccctg tggaatattt cttttttgaa attgtcattt aacttttcct
     3961 ttagtgtctt gtaataagaa attttacttt cattaatcat taaattttgt tcaacataag
     4021 tatctatctt ttttgttaaa actttaacaa cacgtttttt ttccataaaa attttccttt
     4081 atttttgcaa cattttcctt aattgatata attgttacta ttttagattt tttaatatcc
     4141 acttaagaat taatccctca ctagaatatt ttttattata gaattcatat acatgttttt
     4201 ctaaatctat aaatttatca acaattgctt tattataggg tgtatctatc ttttcttttc
     4261 ttaataatcg tctcatattc tttaaataac aacaaacact gccggtttta aacctaaatt
     4321 ctatgtaata tatttttgaa aaagtatata cattttcaac cccgtttatt tcatacttta
     4381 caaaaacatt atttattggt tttctataac catagtaaat gcctaaaaat ttgtccccgt
     4441 ctttcatagg atatagataa agtttttgaa tctctgattt atctaataat ttttcaaaaa
     4501 aaattataca ccgcttttgt ttgttactaa ctactctatg caatgccttt aatatttttg
     4561 tgtgatatat atttttatca cctttttttt ctttcagcag gaagatactt tcttttggaa
     4621 atttaatttt ttgttcattt attcttgcag ccttaagtcc gattttcata tttaatcctt
     4681 atatagcctt tctatttaat tcttttaaac tcaataaact tttttggttt ttcattagtt
     4741 ctaataattc gcaataatat gtgcaaaaca ctttattgta ttctattttc ttttggttat
     4801 tcaaatattc ttttataatt ggctttagaa gttcaatatt tacttctttt tttagttgtt
     4861 caataaggat attgaaaatg tttgttttta aatcttgata attttgttga gaattttctt
     4921 tttttctttc aattgacttc tctaatttac gctttatatt gtttaaatcg ctatatttat
     4981 gactttcaat aataaaatga ggcttaaatt tgtaattttc gtatattttt tggaaacttg
     5041 tttctaattg tttagaatta tattcatttt tttcaaattc tttttgagtg ttgcatagaa
     5101 tttcctttaa tttttcttgt ttttctttca tacaagattt agtaaaatgt atatttttat
     5161 tttttagtaa tttaatttct attattttta tgattttaag tattttgatt atattatctt
     5221 tatcaatatt taaattgaat aaaattgtaa gaaaaattga aagaattttt ttacataaaa
     5281 agttacattt attgaagtaa tttcttactt gatacttttc tatttcatta atctttcttt
     5341 cttcttttat attattttta ttatttaaac actcctttaa atctacactc ccatttttag
     5401 taaatctatc tttaaggccc cttgaaactc tattttgaaa tcttaaatct tttttttctt
     5461 gaaagtattt atagattttt tggtaacatt ctttttttga aaaattaagt tgatagtata
     5521 tttcggtccc caaatttacc cctaagtgtt tgtgataatt agttgtgact ttaaattctt
     5581 tttctaattt gtaaaggtaa ttttgcagtg tttttagttt aacgggtgtt tgactatttc
     5641 ttttaaggtt ttcattaaag taatagagta tgtttttttg tgtgtatttc ttatatgttt
     5701 tatttatgta cgctagtgtt gagataagga ctactaattt atactgtaat ttattgtagc
     5761 aagttgtggt tttactggtt ttgtttatat tagataagtt ttccatttat agctccttga
     5821 ataccggatt ttttatctat aaagtatata tatcacaatt ttgatttaaa agtaaattct
     5881 ttttttaaaa atacaaaaat tttttataaa ttatatagac catgatattg tattaatact
     5941 aattgatttt aatttaaggt tgaactaaaa taaacatagt tttgtagaaa ataatttttc
     6001 attattttgc tacttgatat tggatcgtaa aaatattagg gctttacaaa gttcttttaa
     6061 aagagaattt agtaaagccc taagtctttt aataaaaaat tttttgtaaa aaagttggca
     6121 aaaatagttt ttgctatata tttatatata agaaaaaatt caatactcaa ggagataaaa
     6181 aatggaaaac ttaaacaata atccacaaaa aattaatcaa ataaataagc aaactaattt
     6241 tattggtttt gaaaatttta tagaacttga taatcaagtt aatcaaacta aagaaaaaag
     6301 taaattaagt aagataggta agaaattacc cggaataagc agtcaagtct agtgcggccg
     6361 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6421 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6481 actcattagg caccccaggc
//
