LOCUS       sequence-345220-        9060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of gene cluster mtrCAB from Shewanella 
            oneidensis MR-1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9060)
  AUTHORS   
  TITLE     Turn on the Mtr pathway genes under pLacI promoter in Shewanella 
            oneidensis MR-1
  JOURNAL   Bioresour. Bioprocess. 5, 35 (2018)
REFERENCE   2  (bases 1 to 9060)
  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: "Bioresour. 
            Bioprocess. 5, 35 (2018)"
COMMENT     Sequence Label: pETSXM2-pLacI-mtrCAB
FEATURES             Location/Qualifiers
     source          1..9060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     313..331
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(335..378)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    381..402
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    503..524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(539..544,545..562,563..569)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    577..593
                     /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)."
     RBS             611..622
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      5800..5871
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5887..5914
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    5921..5941
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     rep_origin      7073..7528
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7160..7179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7370..7391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7620..8435)
                     /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(7620..8435)
                     /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     8343..8362
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8427..8446
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(8557..9060,1..85)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(9046..9060,1..5)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctttatagt cctgtcgggt ttcgccactt ctgacttgag cgtcgatttt tgtgatgctc
       61 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      121 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      181 ccgtattacc gcctttgagt gagctgatac cgctcgcact agtcatcaag agatgcggtg
      241 ttgcgtgtgt tgcattctcg agtgccacct gacgtctaag aaaccattat tatcatgaca
      301 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
      361 ttcgctaagg atgatttctg gaattcgcgg ccgcttctag agcaatacgc aaaccgcctc
      421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      481 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      541 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      601 catactagag aaagaggaga aagagctcat gatgaacgca aaaaaatcaa aaatcgcact
      661 gctgctcgca gcaagtgcca tctccatggc cttaactggt tgtggtggta gcgatggtac
      721 cgatggtaac cctggtgagc caggtggaga acccgctggt actatccata aggtaaacct
      781 cgatatcacg aaagtgagct atgaaacggt tcgcctgtgg tcactgtttt cgctaccaac
      841 gaagccgata tgccaattgt gggtctcgca aatttagaaa tcaaaaaagc actgcaatta
      901 ataccggaag gggcgacagg ccctggtaat agtgctaact ggcaaggatt aggctcttca
      961 aagagctatg tcgataataa aaacggtagc tataccttta aattcgacgc cttcgatagc
     1021 aataaagttt ttaatgccca attaacgcaa cggtttaacg ttgtttctgc tgcgggtaag
     1081 ttagcagacg gaacgaccgt ccccgttgcc gaaatggttg aagatttcga cggccaaggc
     1141 aatgctccgc aatacaccaa aaatatcgtt agccacgaag tatgtgcttc ttgccacgtt
     1201 gaaggtgaaa agatttatca ccaagcaact gaagtcgaaa cctgtatatc ttgccacaca
     1261 caagagtttg ctgatggtcg cggtaaaccc catgtcgcct ttagtcactt aattcacaat
     1321 gtgcataatg ccaacaaagc ttggggcaaa aacaataaag tccctgaagt tgcgcaaaaa
     1381 attgtccaag ataattgcca agtgtgtcac attgaatccg acatgcttac cgaggcaaaa
     1441 aactggtcac gagttccaac aatggaagtc tgttctagct gtcacgtaga catcgatttt
     1501 gctgcgggta aaggccactc acaacagctc gataactcaa actgtatcgc ctgtcataag
     1561 agtgattgga cagccgagtt acatacagct aaaaccgctg acaaaaaagc cgttatcgct
     1621 caattaggta tgtctgcaac gttagctgct caagctgata atactgcggt attaaccgtg
     1681 tctatcttag ataaagacgg caatgccatt gatgcggcga gcatacaaaa taagatcaaa
     1741 cgtttagaaa ctatcactaa cgttggtcct aacttcccca tcatgggcta taacccaagc
     1801 ccagtaacac aatataagaa agtgactaaa gacttagtca aggatggcgc actgcaagca
     1861 gatgtcagcg tagttgacgg taaactggtg ttcaccacac ctgccctgcc atttggcacg
     1921 ggcgatactg ataccgcctt tagcttcatc ggtctagaaa tgtgcaatac cggcacagat
     1981 ctaacagctt gtaccgctaa tagcacaacc accagtatga aagccgagtt agcctttggc
     2041 actaagtctg gcaaagcacc aagcatgcgc catgttaact cggttgagtt tgcgacatgc
     2101 caaaactgtc acggcgacac ctttgaaatc cacaaaggcc accatgcagg ctttgtaatg
     2161 tctgaacaag tttcacatgc caaagatgca agcggtaagc ctatcgtcgg cgttgatgct
     2221 tgtgtggcat gccatacacc agacggtacc tatagcccga ctaaactcgg tgcattagag
     2281 atgaaactac acgtggtgca tggtgagcaa ggtatcatta aagaatgtac ccaatgtcat
     2341 aacgacttca atttggatgc atttaaactg aaaggtgcac tagcgacaac aaaaggctat
     2401 accaccccaa tcacagccac ttgtacctca tgtcatacca ttggtacaga taaaatgacc
     2461 aaacattctg tcgacaccct tgtaagctac ggtgctgtta ttgatggtga ttatactcaa
     2521 gccaaccaag cggcacagtc agaaacctgt ttctactgcc ataccccaac agttgcagat
     2581 cacactcaag tgaaaatgta atttgcccaa gcagggggag ttcgctcccc ctttcttgaa
     2641 ttttgttggg acaaattggg aagcctatta tgaagaacag tcacaaaatg aaaaatctac
     2701 tgccggcatt tgtcatcaca atggcaatgt cagcagttat gtctttatgc atcacaccaa
     2761 acgcttacgc gtcgaagtgg gatgaaaaaa tgacccctga gcaagtcgaa gcgaccttgg
     2821 ataagaagtt tgccgaaggc aactactccc ctaaaggtgc agactcttgc atcatgtgcc
     2881 ataaaaaatc tgaaaaagtg atggaccttt tcaaaggtgt ccacggcgcg attgattcgt
     2941 caaaaagccc catggctggc ctgcaatgtg aggcgtgcca cggcccaatg ggtcagcaca
     3001 acaaaggcgg caacgagccg atgatcactt ttggtaagca atcaacctta agtgccgata
     3061 agcaaaacag cgtatgtatg agctgtcacc aagacgataa acgtatgtct tggaatggcg
     3121 gtcaccatga caacgccgat gttgcttgtg cttcttgtca ccaagtacac gtagctaaag
     3181 atcctgtgtt atctaaaaac actgagatgg aagtctgtac tagctgccac actaagcaaa
     3241 aagcggatat gaataaacgc tcaagtcatc cactcaaatg ggcccaaatg acctgtagcg
     3301 actgccacaa tccccatggg agcatgacag attccgatct taacaagcct agtgttaatg
     3361 atacctgtta ttcctgtcat gccgagaaac gcggcccaaa actttgggag catgcacccg
     3421 tcactgagaa ttgtgtcact tgccacaatc ctcacggtag tgtgaatgat gccatgctga
     3481 aaacccgcgc gccgcagctg tgtcagcaat gtcatgctag cgatggccac gccagcaatg
     3541 cttacttagg taacactggt ttaggttcta atgtcggtga caatgccttt acgggtggaa
     3601 gaagctgctt aaattgccat agtcaggtcc atggttctaa ccatccatct ggcaagctat
     3661 tacagcgcta aggagacgag aaaatgaaat ttaaactcaa tttgatcact ctagcgttat
     3721 tagccaacac tggctttgcc gtagccgctg atggttatgg tcttgctaat gccaatactg
     3781 ataaagtgaa attatccgca tggagctgca aaggttgcgt ggttgaaaca ggcacatctg
     3841 gcactgtggg tgtcggtgtc ggttataaca gcgaagagga tattcgctca gccaatgcct
     3901 ttggaacatc caatgaagtc gcgggcaaat ttgatgccga tttaagcttt aaaggtgaaa
     3961 aaggttaccg tgccagtgtt gaagcttatc aactcggtat ggatggcggt cgtttagatg
     4021 tgaatgcggg caaacaaggc cagtacaacg tcaatgtaaa ctatcgtcaa attgcaacct
     4081 acgacagcaa tagcgcccta tcgccctacg cgggtattgg tggcaataac ctgacgttac
     4141 ccgataactg ggtaactgct ggttcaagca gccaaatgcc attgttgatg gatagcctca
     4201 atgcccttga attatcactt aaacgtgaac gcacgggatt gggatttgaa tatcaaggcg
     4261 aatccctgtg gagcacctat gttaactaca tgcgtgaaga aaaaactggc ttaaaacaag
     4321 cctctggtag cttcttcaac caatcgatga tgttagcaga gccagtggat tacactactg
     4381 acaccattga agcgggtgtc aaactcaagg gtgatcgttg gttcacagcg ctcagttaca
     4441 atgggtctat attcaaaaac gaatacaacc aattggactt tgatagtgca ttcaatccta
     4501 cctttggtgc ccaaacccaa ggtacgttgg cgcttgatcc cgataatcag gcacacacag
     4561 tgtcattgat gggacagtac aacgatggca gtaacgccct atcgggtcgt attcttactg
     4621 gacaaatgag ccaagatcag gcgttagtaa cggataacta tcgttattcc agccagctaa
     4681 caagtgatgc ggttgatgca aaagtcgatc tattgggcat gaaccttaaa gtcgtcagca
     4741 aagtcagcaa agatcttcgc ttgacaggta gttacgatta ttacgaccgt gacaataata
     4801 cccaagttga agaatggacc cagatcagca tcaacaatgt caacggtaag gtggcttata
     4861 acacgcctta tgataatcgt acgcaacgct ttaaagtcgc ggcagattat cgtattaccc
     4921 gcgatattaa acttgatggt ggttatgact tcaaacgtga ccaacgtgat tatcaagacc
     4981 gtgaaaccac ggatgaaaat accgtttggg ctcgtttacg cgtcaatagc ttcgatatgt
     5041 gggacatgtg ggtaaaaggc agttacggaa aacgtgacgg ctcacaatac caagcatctg
     5101 aatggacttc ttctgaaacc aacagcctgt tacgtaagta caatctggct gaccgtgaca
     5161 gaacccaagt cgaagcgcgc atcacccatt cgccgcttga aagcctgacc atcgatgttg
     5221 gtgcccgtta cgcgttggat gattataccg atactgtgat tggcttaact gagtcaaaag
     5281 acaccagtta tgatgccaat atcagctata tgatcaccgc tgacttactg gcaaccgcct
     5341 tctacaacta tcaaaccatt gagtctgaac aagcaggtag tagcaattac agcactccaa
     5401 cctggactgg ctttattgaa gacaaggtcg atgtagttgg cgcaggtatt agctacaaca
     5461 atctgctaga gaacaagtta cgcctcggac tggactacac ctattccaac tccgacagta
     5521 acactcaagt cagacaaggt atcactggcg actatggcga ttattttgcc aaagtacata
     5581 acattaactt atatgctcaa tatcaagcca ccgaaaaact cgcgctgcgc ttcgattaca
     5641 aagttgagaa ctataaggac aatgacgccg caaatgatat cgccgttgat ggcatttgga
     5701 acgtcgtagg ttttggtact aacaaccatg actacaccgc acaaatgcta atgctgagta
     5761 tgagctacaa gctctaacat atgtactaga gccaggcatc aaataaaacg aaaggctcag
     5821 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     5881 cacactggct caccttcggg tgggcctttc tgcgtttata tactagtagc ggccgctgca
     5941 gccatgggtc gactgattat acgtctaatt ttagctgtgc gctctgcttg aactcaaatg
     6001 acaatgcagc cactgttcgc cctttcttaa tggtctcgta atttatggtg aggtctgaac
     6061 gctgatttag ttcatctacg gccggcttga taacccattt atttaagtct ctaaattgtg
     6121 ggtatttgtc ggagatacct aatgctgagc ggaagtcatc cacatagata actctttctc
     6181 cagtagcgtt aaatttcatc aaaagttcgt ataaacgaat agaatgagct gactgtagat
     6241 tcgcaatatg tcgcagctta taagttgtaa acctgctctg taaactactg atgtacttca
     6301 aaacttcatc agtccaagtg attgttattg ctccttgtcc tttgagtttt ttcactcctt
     6361 tttgaatcca cctaagcttt acctcttctt gctcatctct taggagtatt acggcatcga
     6421 aaagggtatc agacgccttg tatagctctc tatgagcatt tttgatgtct atccccataa
     6481 gttctgagaa ttcaagggct gttagggtca tttcctttgg tgcattacct ctagaatcta
     6541 ctttcccgag gcaagctaag accagttttt gagcttgtac atttagtttg taactagcat
     6601 ctatcaagct atttgattta gttacagata ggttgttcat tttacgtcac tattgtggag
     6661 tttatttttg gggattccac aataaccaat tgtggaattg ctatcaacgt tctccacaat
     6721 taactgtgaa tatcaccgta aatctctcca cctttaggga ttgttttacc gtaaatctct
     6781 ccacctttag gggctgtttt accgtaaatc tctccacatt aaaccgctgt agccctttag
     6841 caccaagggc tacagcggcg ctaaaaacaa gaaagatctt aaaaaacaag aaagatctat
     6901 taaaaagggt ttcgcctgtg gataaacaag taaatttgag catctggcgg ctgcgccgcg
     6961 accaagggct tcgccctagt tcgcctaaag gctcggatcc cttggggcct ctaaacgggt
     7021 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     7081 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     7141 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     7201 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     7261 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     7321 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7381 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     7441 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7501 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     7561 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
     7621 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     7681 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     7741 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     7801 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     7861 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     7921 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     7981 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgag
     8041 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatac
     8101 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     8161 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     8221 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     8281 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     8341 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     8401 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     8461 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8521 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8581 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggttagtt tgccggatca
     8641 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8701 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8761 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8821 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8881 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8941 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9001 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
//
