LOCUS       sequence-378887-        9977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacteroides - Escherichia coli shuttle vector with TetQ and 
            Kanamycin selection markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9977)
  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 9977)
  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: pG108 - K
FEATURES             Location/Qualifiers
     source          1..9977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     219..238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             4986..4994
                     /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     5401..5418
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5535..5556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5571..5576,5577..5594,5595..5601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5609..5625
                     /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     5614..5636
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5633..5649
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5633..5649
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5645..5968
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    5662..5718
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5719..5736)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5719..5735)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5728..5750)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5944..5963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(8770..9585)
                     /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(8770..9585)
                     /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     9493..9512
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9577..9596
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(9707..9977,1..318)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
       61 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      121 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      181 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      241 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      301 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      361 tggccttttg ctcacatgtg atcctctatt gcagatggtc ttgttgcaca cccctctgaa
      421 gggttctata tcatctatac cacctcttca gaggagtttc tcacgttatc tgaagaagat
      481 tattctctgt tcgcttcttt tgccaatttg gttgttcgag aggactccaa ttcaaatggc
      541 ttaagaagcg caaaaccctc tggggatggc tggaaatttg gaggtaaagg aaaggggaaa
      601 ctagatgcca tccacatcgg ctatgaatta tcgaaaatcc taggagaaaa tcaagatttt
      661 gagatctatg tagaatacaa tggcgatggg tcttataccg tttggtatag ggaagtccaa
      721 aaatcgtcta aagggagtaa tccgtaaaga cttttaatgt cctcataaga gccaacgaaa
      781 gagaggggca tacgtctctc tctttcaacg tatgcttgaa cgattgctta ctcgatgtga
      841 ttaaatagag ctggcttgat aggtcagctc tatttctgtt tttcgttttg aaaatatgat
      901 aggcaaatac catggatatg acataacaga aatagagcta ctaagcccaa agccacgaac
      961 cataacgatg tcagaggatc acgtggcgct acttagtagc tctatctcac ttgcaaagtt
     1021 actaaaagag atctgcgtgc gagcctaagc gtgagaggac gataagctcc tcgtctgcgc
     1081 ttatctgctc tatccatatc aagagcaggt caccctcaat gtgtagctcc atacagccct
     1141 catagttccc tataagcttg tgaggacgca tttctaaggg tatctccaca cccttagaca
     1201 gcatttcaat gacatctacg attttagagc gccttttgag gtcgttccta aatcgcttaa
     1261 ggtctttctt aaacttggta gtatatacca gcctacgctt cctcatttcc aaacaaagac
     1321 gcttctattg tttctcgggt agttgtatcc acagagccgg catagcgacc gctcttagcg
     1381 tctatgcagg cctctgccgt ctcggcatta taaggtctat accccattcg gtagagtaag
     1441 gtctcaatat agttgcttag gcttctattc tcagcccttg cttcactgcg tatctgctgg
     1501 agtacactag catcaattct aattgacgtg gatgctttta gctcaatatt atccatttct
     1561 attatattct acttgattat attcactgca aatatagaca aatctcccaa ataaacgtct
     1621 gtcggctggg cgtcagttca cgcaccaaat gtgtgtgtaa tgggagcgac aagaagttgt
     1681 ccaataaaat cgttctccgt aactttgggc aggcgctagc tacgaccata gttatagcac
     1741 attataacta catacgtagt tgtgtgctct ccgaggggca tggaggacac ccccagcacc
     1801 ccctaaagtt gacccaaaag tacaagcgat gagcaaaaca agtatccata tcgagccgtg
     1861 taaaatcggc agtagtgagc aacataatca acgcttgaag catcttgatt atgtgcgacc
     1921 cgacctctcc ccacaaaacg agagttgggt tggcgtagca gatttgcccg cgcatcttga
     1981 acaactccga atacttgtaa aggagaagac gggcagagcg atgcaacaga aagcaacccc
     2041 gataagagag ggtgtaatag taatcaatca aggcacctcg attaaccaat taaggggtct
     2101 cgcaacggca atagaactgc gatggggtat taagacgcta cagatataca cccacaagga
     2161 cgaggggcat atagatagcg atgacacgtg gaagccaaac ctacatgccc atatggtgct
     2221 tgattgggtg aaccacgaca cgggcaagag tatcaagctt agcaagcaag atatggcgga
     2281 gatgcagacg atggtggcgg actgcctgca aatggttagg ggagagagta gcgatgtcaa
     2341 acacctcggg gctatacaat acaagaatca agccgaagag ctgcgtctcc aaacccttaa
     2401 agaacaaacg atgaaagaag aacaggctcg ggagctggca ctagccgaag ctaagcggat
     2461 ggagcaggtt gcgtctgagc agattatcgc agagaataag aagcttggag ggctgtggat
     2521 ggactggcaa aaatcatttg aggcctctga gaagcgccga gagagcataa cggctcgtaa
     2581 gatagaggag gataagctat gggaagaact tgggttgaaa ggagccttta aatcgttttg
     2641 ggaggaatgc aaggattggt ttagaggttt cggcgatggg ctcaaagact tagctctgct
     2701 gatatgggct tcgcctagag gagtgatgga caaagacgtg aggtgcacag cggatcggaa
     2761 gagggggaaa ttactgctca atgggcatac actagagcag gtaagagcac ccaagatagt
     2821 tgtaccaagc tcccccatac aaaggcaggc gcaatccgtc agtgagtgcg ctacgaggca
     2881 ggctagagat acagacagac gcagagagaa gcgtgttgac caagagagac gagagcgcag
     2941 gggtcgcaag atttgaccgc actatgttct taaagcgttt cccaagggct ttgccaccac
     3001 ttgcggacgg tcggtctact tagtcctgtc tctctcgcac aacggctctt gttgttgctg
     3061 tttgggtttc tatccctcca ctcctgcacc ttggcgtatt gcgggctgtc ttccgctagt
     3121 acgatagagc cttttggtct tccgttaccc tctctccaat cttttcttcc ctttgcttcc
     3181 tgcaaaatat ctctagagcc tcttgcgagc tttagatgta agtcttgcgg tctcccattg
     3241 cgtttgttag gttctattct tagccccgac ctatcctcta tcagctggcg agggtatgtc
     3301 tctgagctct ccttatatgc ttttagagcg tctagagcgt cctgcacgga gaagtgcctg
     3361 cccgtgttgc ccgttagtgc ttccattcgt ggcacaagct ctagagcgtc ccgtttgagt
     3421 atgtcaaatg gcacgttaca cttcttggca tatactgcta agaacataat gcagtggtag
     3481 cgatgcccaa cagcgacctt ttcgttcgtc cacagcatat gtagccacca ctcatacagc
     3541 cccctatgca catgccacgt cttacgaggt gcgccctcta cgatgcgttt ttgataccaa
     3601 tctgggtata gctctttagc tcttgctttt gttagcctat tcgtggagta tgcccctttg
     3661 tccaacgctt gcgcctgctc ctgcgacacc cccgagttaa ggtacttcag tgtaggcttg
     3721 ctagcaaacc gattcagctc ctcaatagtg tgtagtgggg catcagcatt gtgccaagct
     3781 gttacaatgt ccccgtgctt cgtacgactg cctggcaccc taaacggctg tataatgggc
     3841 tgtatttgca cttggttgtc ctcccccgtc ctagaggacg ctttgttcca tataagccgt
     3901 gtaagagctt ttttaatgtt ggtgagcgtc tcgtacacct tggggtataa gggcatcggc
     3961 ttagcaagta tgtagtacaa gtggatgcca tcgcccgagt tgacgatgat attagcttgt
     4021 ggggtcatcc ctatcgtctg ctgatggatt acctctgaca tctctggcac tccgacctca
     4081 tcaaggtcaa aggcgagggc atacagatag cgggcattgc tcttgaacct cctgcgcccc
     4141 acgtaggtgg ctggggttgt taccacaaac ttgtaccgcc ccatcagctc ctctaagcta
     4201 tcccaaccac acaccttgat gtcatcctcg cttgtggcat cgcccactac cacccacgtc
     4261 ttagacctcc tgccattatc attagccctg cctgcatcag gctctacgct gtagacagct
     4321 tgcaccaaag cattatagcg gaaaggttga gctggattat agattgcttt cttacctgct
     4381 ttggacttag actttctccc tgcttgttga gccctccaat cgatgtattc ttggcacagc
     4441 tctttttcaa aaggatagat agggcattgc gggtcattat ggtcacaaga accaaagatt
     4501 atttggaaga actcaaaggg tttaagagat cgatattcaa aggtctcatt actctctaag
     4561 tataaatttt tagagagaaa aacatcgtga gaaatattag aattatcagt catcgttcta
     4621 cgtcattttg gggtgatttt ttggtgtcgc ttcgctcccc caaaaaatca tacggggggc
     4681 atatattatc aatatatttt ttttgccact tgtggcacaa agataaggag gatttttgag
     4741 acctccttct acgtcatttt ggggtgattt tttggtgtcg cttcgctccc ccaaaaaatc
     4801 atacgggggg catatattat caatatattt ttttttgcca cttgtggcac aaagataagg
     4861 aggatttttg agacctcctt ctacgtcatt ttgggtggaa tttttggctc gctacgctcc
     4921 ccaaaaattc atacgggggg gcgtatatta tcaatgaagt aaaatcccca cttgtggtac
     4981 aaaggtaagg aggttttttg agacctcctt ctgcgtcatc gttctacgtc attttggggt
     5041 tgattttttg gggaagaaac aaagaatatc gcaacaagta ggtattgcat atattttttg
     5101 caattcttta tctttgcggt aagacaacct cttaagagag acttatcttg aatacaaaac
     5161 actaaaccta agcagctatg aaaactagaa cattctgcgc aatagcaatc gggctattgt
     5221 ctttggcgtt aacttcgtgc caaaacaacc ttgacttgtt aaagccaagc caagacactg
     5281 aaccagaaca aatcactcaa ttacatgttc tttcctgcgt tatcccctga ttctgtggat
     5341 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5401 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5461 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5521 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     5581 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5641 agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg
     5701 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5761 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5821 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     5881 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     5941 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6001 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6061 ctccgggagt ccatcactcc tctaggcgaa gcccatatca gcagagctaa gacgtcacca
     6121 gttgaaccta cgtttcccta ataaaatgtc tatggtaaaa agttataaaa tcctcctact
     6181 tttgttagat atattttttt gtgtaatttt gtaatcgtta tgcggcagta ataatataca
     6241 tattaatacg agttaggaat cctgtagttc tcatatgcta cgaggaggta ttaaaaggtg
     6301 cgtttcgaca atgcatctat tgtagtatat tattgcttaa tccaaatgaa tattataaat
     6361 ttaggaattc ttgctcacat tgatgcagga aaaacttccg taaccgagaa tctgctgttt
     6421 gccagtggag caacggaaaa gtgcggctgt gtggataatg gtgacaccat aacggactct
     6481 atggatatag agaaacgtag aggaattact gttcgggctt ctacgacatc tattatctgg
     6541 aatggtgtga aatgcaatat cattgacact ccgggacaca tggattttat tgcggaagtg
     6601 gagcggacat tcaaaatgct tgatggagca gtcctcatct tatccgcaaa ggaaggcata
     6661 caagcgcaga caaagttgct gttcaatact ttacagaagc tgcaaatccc gacaattata
     6721 tttatcaata agattgaccg agccggtgtg aatttggagc gtttgtatct ggatataaaa
     6781 gcaaatctgt ctcaagatgt cctgtttatg caaaatgttg tcgatggatc ggtttatccg
     6841 gtttgctccc aaacatatat aaaggaagaa tacaaagaat ttgtatgcaa ccatgacgac
     6901 aatatattag aacgatattt ggcggatagc gaaatttcac cggctgatta ttggaatacg
     6961 ataatcgctc ttgtggcaaa agccaaagtc tatccggtgc tacatggatc agcaatgttc
     7021 aatatcggta tcaatgagtc gttggacgcc atcacttctt ttatacttcc tccggcatcg
     7081 gtctcaaaca gactttcgtc ttatctttat aagatagagc atgaccccaa aggacataaa
     7141 agaagttttc taaaaataat tgacggaagt ctgagacttc gagacgttgt aagaatcaac
     7201 gattcggaaa aattcatcaa gattaaaaat ctaaaaacta tcaatcaggg cagagagata
     7261 aatgttgatg aagtgggcgc caatgatatc gcgattgtag aggatatgga tgattttcga
     7321 atcggaaatt atttaggtgc tgaaccttgt ttgattcaag gattatcgca tcagcatccc
     7381 gctctcaaat cctccgtccg gccagacagg cccgaagaga gaagcaaggt gatatccgct
     7441 ctgaatacat tgtggattga agacccgtct ttgtcctttt ccataaactc atatagtgat
     7501 gaattggaaa tctcgttata tggtttaacc caaaaggaaa tcatacagac attgctggaa
     7561 gaacgatttt ccgtaaaggt ccattttgat gagatcaaga ctatatacaa agaacgacct
     7621 gtaaaaaagg tcaataagat tattcagatc gaagtgccgc ccaaccctta ttgggccaca
     7681 atagggctga ctcttgaacc cttaccgtta gggacagggt tgcaaatcga aagtgacatc
     7741 tcctatggtt atctgaacca ttcttttcaa aatgccgttt ttgaagggat tcgtatgtct
     7801 tgccaatccg ggttacatgg atgggaagtg actgatctga aagtaacttt tactcaagcc
     7861 gagtattata gcccggtaag tacacctgct gatttcagac agctgacccc ttatgtcttc
     7921 aggctggcct tgcaacagtc aggtgtggac attctcgaac cgatgctcta ttttgagttg
     7981 cagatacccc aagcggcaag ttccaaagct attacagatt tgcaaaaaat gatgtctgag
     8041 attgaagaca tcagttgcaa taatgagtgg tgtcatatta aagggaaagt tccattaaat
     8101 acaagtaaag actatgcatc agaagtaagt tcatacacta agggcttagg catttttatg
     8161 gttaagccat gcgggtatca aataacaaaa ggcggttatt ctgataatat ccgcatgaac
     8221 gaaaaagata aacttttatt catgttccaa aaatcaatgt catcaaaata atggagcggt
     8281 caggaaattt ctataaggca atacagttgg gatatatact tatctccatt cttatcggat
     8341 gtatggcata taatagcctc tatgaatggc aggagagacg tcaggtggca cttttcgggg
     8401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     8461 catgagacaa taaccctgat aaatgcttca ataataattc gagctcggta cccccgatag
     8521 cttccgctat tgcttttttg ctcatcggta tttgcaacat catagaaatt gcataccttt
     8581 gttcctcggt tatatgtttg ctcatctgca actttttttt ctttggacgg acaattaaag
     8641 caaagatagc aaactttatc cattcagagt gagagaaagg gggacattgt ctctctttca
     8701 aatgctggaa gttgtccctg aaaaatttca tccttcgtag ttcaaagtcg ggtggttgtc
     8761 aagatgttct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     8821 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     8881 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     8941 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     9001 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     9061 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     9121 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     9181 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     9241 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     9301 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     9361 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     9421 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     9481 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     9541 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     9601 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     9661 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     9721 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     9781 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     9841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     9901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     9961 agtcgtgtct taccggg
//
