LOCUS       sequence-378890-        9084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacteroides - Escherichia coli shuttle vector with ErmF and Kan 
            selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9084)
  AUTHORS   Jones KR, Belvin BR, Macrina FL, Lewis JP
  TITLE     Sequence and characterization of shuttle vectors for molecular 
            cloning in Porphyromonas, Bacteroides and related bacteria.
  JOURNAL   Mol Oral Microbiol. 2020 Aug;35(4):181-191. doi: 10.1111/omi.12304. 
            Epub 2020 Jul 9.
  PUBMED    32592236
REFERENCE   2  (bases 1 to 9084)
  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"; doi: "10.1111/omi.12304";
            journalName: "Mol Oral Microbiol"; date: "2020-08"; volume: "35"; 
            issue: "4"; pages: "181-191"
COMMENT     Sequence Label: pG106K
FEATURES             Location/Qualifiers
     source          1..9084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     218..237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             4985..4993
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     5400..5417
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5534..5555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5570..5575,5576..5593,5594..5600)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5608..5624
                     /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     5613..5635
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5632..5648
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5632..5648
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5644..5967
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    5661..5717
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5718..5735)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5718..5734)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5727..5749)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5943..5962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     oriT            6214..6323
                     /note="incP origin of transfer"
     oriT            6471..6580
                     /note="incP origin of transfer"
     CDS             complement(7876..8691)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7876..8691)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     8599..8618
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8683..8702
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(8813..9084,1..317)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
       61 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      361 ggccttttgc tcacatgtga tcctctattg cagatggtct tgttgcacac ccctctgaag
      421 ggttctatat catctatacc acctcttcag aggagtttct cacgttatct gaagaagatt
      481 attctctgtt cgcttctttt gccaatttgg ttgttcgaga ggactccaat tcaaatggct
      541 taagaagcgc aaaaccctct ggggatggct ggaaatttgg aggtaaagga aaggggaaac
      601 tagatgccat ccacatcggc tatgaattat cgaaaatcct aggagaaaat caagattttg
      661 agatctatgt agaatacaat ggcgatgggt cttataccgt ttggtatagg gaagtccaaa
      721 aatcgtctaa agggagtaat ccgtaaagac ttttaatgtc ctcataagag ccaacgaaag
      781 agaggggcat acgtctctct ctttcaacgt atgcttgaac gattgcttac tcgatgtgat
      841 taaatagagc tggcttgata ggtcagctct atttctgttt ttcgttttga aaatatgata
      901 ggcaaatacc atggatatga cataacagaa atagagctac taagcccaaa gccacgaacc
      961 ataacgatgt cagaggatca cgtggcgcta cttagtagct ctatctcact tgcaaagtta
     1021 ctaaaagaga tctgcgtgcg agcctaagcg tgagaggacg ataagctcct cgtctgcgct
     1081 tatctgctct atccatatca agagcaggtc accctcaatg tgtagctcca tacagccctc
     1141 atagttccct ataagcttgt gaggacgcat ttctaagggt atctccacac ccttagacag
     1201 catttcaatg acatctacga ttttagagcg ccttttgagg tcgttcctaa atcgcttaag
     1261 gtctttctta aacttggtag tatataccag cctacgcttc ctcatttcca aacaaagacg
     1321 cttctattgt ttctcgggta gttgtatcca cagagccggc atagcgaccg ctcttagcgt
     1381 ctatgcaggc ctctgccgtc tcggcattat aaggtctata ccccattcgg tagagtaagg
     1441 tctcaatata gttgcttagg cttctattct cagcccttgc ttcactgcgt atctgctgga
     1501 gtacactagc atcaattcta attgacgtgg atgcttttag ctcaatatta tccatttcta
     1561 ttatattcta cttgattata ttcactgcaa atatagacaa atctcccaaa taaacgtctg
     1621 tcggctgggc gtcagttcac gcaccaaatg tgtgtgtaat gggagcgaca agaagttgtc
     1681 caataaaatc gttctccgta actttgggca ggcgctagct acgaccatag ttatagcaca
     1741 ttataactac atacgtagtt gtgtgctctc cgaggggcat ggaggacacc cccagcaccc
     1801 cctaaagttg acccaaaagt acaagcgatg agcaaaacaa gtatccatat cgagccgtgt
     1861 aaaatcggca gtagtgagca acataatcaa cgcttgaagc atcttgatta tgtgcgaccc
     1921 gacctctccc cacaaaacga gagttgggtt ggcgtagcag atttgcccgc gcatcttgaa
     1981 caactccgaa tacttgtaaa ggagaagacg ggcagagcga tgcaacagaa agcaaccccg
     2041 ataagagagg gtgtaatagt aatcaatcaa ggcacctcga ttaaccaatt aaggggtctc
     2101 gcaacggcaa tagaactgcg atggggtatt aagacgctac agatatacac ccacaaggac
     2161 gaggggcata tagatagcga tgacacgtgg aagccaaacc tacatgccca tatggtgctt
     2221 gattgggtga accacgacac gggcaagagt atcaagctta gcaagcaaga tatggcggag
     2281 atgcagacga tggtggcgga ctgcctgcaa atggttaggg gagagagtag cgatgtcaaa
     2341 cacctcgggg ctatacaata caagaatcaa gccgaagagc tgcgtctcca aacccttaaa
     2401 gaacaaacga tgaaagaaga acaggctcgg gagctggcac tagccgaagc taagcggatg
     2461 gagcaggttg cgtctgagca gattatcgca gagaataaga agcttggagg gctgtggatg
     2521 gactggcaaa aatcatttga ggcctctgag aagcgccgag agagcataac ggctcgtaag
     2581 atagaggagg ataagctatg ggaagaactt gggttgaaag gagcctttaa atcgttttgg
     2641 gaggaatgca aggattggtt tagaggtttc ggcgatgggc tcaaagactt agctctgctg
     2701 atatgggctt cgcctagagg agtgatggac aaagacgtga ggtgcacagc ggatcggaag
     2761 agggggaaat tactgctcaa tgggcataca ctagagcagg taagagcacc caagatagtt
     2821 gtaccaagct cccccataca aaggcaggcg caatccgtca gtgagtgcgc tacgaggcag
     2881 gctagagata cagacagacg cagagagaag cgtgttgacc aagagagacg agagcgcagg
     2941 ggtcgcaaga tttgaccgca ctatgttctt aaagcgtttc ccaagggctt tgccaccact
     3001 tgcggacggt cggtctactt agtcctgtct ctctcgcaca acggctcttg ttgttgctgt
     3061 ttgggtttct atccctccac tcctgcacct tggcgtattg cgggctgtct tccgctagta
     3121 cgatagagcc ttttggtctt ccgttaccct ctctccaatc ttttcttccc tttgcttcct
     3181 gcaaaatatc tctagagcct cttgcgagct ttagatgtaa gtcttgcggt ctcccattgc
     3241 gtttgttagg ttctattctt agccccgacc tatcctctat cagctggcga gggtatgtct
     3301 ctgagctctc cttatatgct tttagagcgt ctagagcgtc ctgcacggag aagtgcctgc
     3361 ccgtgttgcc cgttagtgct tccattcgtg gcacaagctc tagagcgtcc cgtttgagta
     3421 tgtcaaatgg cacgttacac ttcttggcat atactgctaa gaacataatg cagtggtagc
     3481 gatgcccaac agcgaccttt tcgttcgtcc acagcatatg tagccaccac tcatacagcc
     3541 ccctatgcac atgccacgtc ttacgaggtg cgccctctac gatgcgtttt tgataccaat
     3601 ctgggtatag ctctttagct cttgcttttg ttagcctatt cgtggagtat gcccctttgt
     3661 ccaacgcttg cgcctgctcc tgcgacaccc ccgagttaag gtacttcagt gtaggcttgc
     3721 tagcaaaccg attcagctcc tcaatagtgt gtagtggggc atcagcattg tgccaagctg
     3781 ttacaatgtc cccgtgcttc gtacgactgc ctggcaccct aaacggctgt ataatgggct
     3841 gtatttgcac ttggttgtcc tcccccgtcc tagaggacgc tttgttccat ataagccgtg
     3901 taagagcttt tttaatgttg gtgagcgtct cgtacacctt ggggtataag ggcatcggct
     3961 tagcaagtat gtagtacaag tggatgccat cgcccgagtt gacgatgata ttagcttgtg
     4021 gggtcatccc tatcgtctgc tgatggatta cctctgacat ctctggcact ccgacctcat
     4081 caaggtcaaa ggcgagggca tacagatagc gggcattgct cttgaacctc ctgcgcccca
     4141 cgtaggtggc tggggttgtt accacaaact tgtaccgccc catcagctcc tctaagctat
     4201 cccaaccaca caccttgatg tcatcctcgc ttgtggcatc gcccactacc acccacgtct
     4261 tagacctcct gccattatca ttagccctgc ctgcatcagg ctctacgctg tagacagctt
     4321 gcaccaaagc attatagcgg aaaggttgag ctggattata gattgctttc ttacctgctt
     4381 tggacttaga ctttctccct gcttgttgag ccctccaatc gatgtattct tggcacagct
     4441 ctttttcaaa aggatagata gggcattgcg ggtcattatg gtcacaagaa ccaaagatta
     4501 tttggaagaa ctcaaagggt ttaagagatc gatattcaaa ggtctcatta ctctctaagt
     4561 ataaattttt agagagaaaa acatcgtgag aaatattaga attatcagtc atcgttctac
     4621 gtcattttgg ggtgattttt tggtgtcgct tcgctccccc aaaaaatcat acggggggca
     4681 tatattatca atatattttt tttgccactt gtggcacaaa gataaggagg atttttgaga
     4741 cctccttcta cgtcattttg gggtgatttt ttggtgtcgc ttcgctcccc caaaaaatca
     4801 tacggggggc atatattatc aatatatttt tttttgccac ttgtggcaca aagataagga
     4861 ggatttttga gacctccttc tacgtcattt tgggtggaat ttttggctcg ctacgctccc
     4921 caaaaattca tacggggggg cgtatattat caatgaagta aaatccccac ttgtggtaca
     4981 aaggtaagga ggttttttga gacctccttc tgcgtcatcg ttctacgtca ttttggggtt
     5041 gattttttgg ggaagaaaca aagaatatcg caacaagtag gtattgcata tattttttgc
     5101 aattctttat ctttgcggta agacaacctc ttaagagaga cttatcttga atacaaaaca
     5161 ctaaacctaa gcagctatga aaactagaac attctgcgca atagcaatcg ggctattgtc
     5221 tttggcgtta acttcgtgcc aaaacaacct tgacttgtta aagccaagcc aagacactga
     5281 accagaacaa atcactcaat tacatgttct ttcctgcgtt atcccctgat tctgtggata
     5341 accgtattac cgcctttgag tgagctgata ccgctcgtcg cagccgaacg accgagcgca
     5401 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5461 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5521 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5581 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5641 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg
     5701 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5761 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5821 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     5881 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5941 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6001 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6061 tccgggagtc catcactcct ctaggcgaag cccatatcag cagagctaag acgtcgtctg
     6121 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     6181 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     6241 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     6301 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     6361 gaacccgacg tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg
     6421 cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc
     6481 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
     6541 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
     6601 ccaaggaaag tctacacgaa cccgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6661 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6721 ccctgataaa tgcttcaata ataattcgag ctcggtaccc ccgatagctt ccgctattgc
     6781 ttttttgctc atcggtattt gcaacatcat agaaattgca tacctttgtt cctcggttat
     6841 atgtttgctc atctgcaact tttttttctt tggacggaca attaaagcaa agatagcaaa
     6901 ctttatccat tcagagtgag agaaaggggg acattgtctc tctttcctct ctgaaaaata
     6961 aatgttttta ttgcttatta tccgcaccca aaaagttgca tttataagtt gaactcaaga
     7021 agtattcacc tgtaagaagt tactaatgac aaaaaagaaa ttgcccgttc gttttacggg
     7081 tcagcacttt actattgata aagtgctaat aaaagatgca ataagacaag caaatataag
     7141 taatcaggat acggttttag atattggggc aggcaagggg tttcttactg ttcatttatt
     7201 aaaaatcgcc aacaatgttg ttgctattga aaacgacaca gctttggttg aacatttacg
     7261 aaaattattt tctgatgccc gaaatgttca agttgtcggt tgtgatttta ggaattttgc
     7321 agttccgaaa tttcctttca aagtggtgtc aaatattcct tatggcatta cttccgatat
     7381 tttcaaaatc ctgatgtttg agagtcttgg aaattttctg ggaggttcca ttgtccttca
     7441 attagaacct acacaaaagt tattttcgag gaagctttac aatccatata ccgttttcta
     7501 tcatactttt tttgatttga aacttgtcta tgaggtaggt cctgaaagtt tcttgccacc
     7561 gccaactgtc aaatcagccc tgttaaacat taaaagaaaa cacttatttt ttgattttaa
     7621 gtttaaagcc aaatacttag catttatttc ctgtctgtta gagaaacctg atttatctgt
     7681 aaaaacagct ttaaagtcga ttttcaggaa aagtcaggtc aggtcaattt cggaaaaatt
     7741 cggtttaaac cttaatgctc aaattgtttg tttgtctcca agtcaatggt taaactgttt
     7801 tttggaaatg ctggaagttg tccctgaaaa atttcatcct tcgtagttca aagtcgggtg
     7861 gttgtcaaga tgttcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     7921 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     7981 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     8041 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     8101 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     8161 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     8221 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     8281 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     8341 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     8401 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     8461 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     8521 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     8581 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     8641 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     8701 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     8761 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     8821 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8881 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8941 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     9001 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     9061 gcgataagtc gtgtcttacc gggt
//
