LOCUS       sequence-323851-        6442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli Nissle pMUT1-derived plasmid with tetracycline resistance 
            and constitutive RFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6442)
  AUTHORS   Kan A, Gelfat I, Emani S, Praveschotinunt P, Joshi NS
  TITLE     Plasmid Vectors for in Vivo Selection-Free Use with the Probiotic E.
            coli Nissle 1917.
  JOURNAL   ACS Synth Biol. 2020 Dec 10. doi: 10.1021/acssynbio.0c00466.
  PUBMED    33301298
REFERENCE   2  (bases 1 to 6442)
  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: "ACS Synth 
            Biol. 2020 Dec 10. doi: 10.1021/acssynbio.0c00466."
COMMENT     Sequence Label: pM1s3TsR
FEATURES             Location/Qualifiers
     source          1..6442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(172..201)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    248..269
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        join(283..288,289..305,306..311)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             359..1549
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVVGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(711..730)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      complement(1679..1708)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        1790..1824
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1833..1844
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1851..2528
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2573..2644
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2660..2687
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2694..2714
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2750..2781
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             3093..3110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3163..3181)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3177..3224
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5125..5144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5367..5384
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccctaaatct agaatgcggt cgagctcgta aacttggtct gacagctcta gctccggcaa
       61 aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
      121 cgtttgccac ctgacgtcta agaaaaggaa tattcagcaa tttgcccgtg ccgaagaaag
      181 gcccacccgt gaaggtgagc cagtgagttg attgctacgt aattagttag ttagccctta
      241 gtgactcgaa ttcgcggccg cttctagagg agattctcat gtttgacagc ttatcatcga
      301 taagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat
      361 gaaatctaac aatgcgctca tcgtcattct cggcaccgtc accctggacg ctgtaggcat
      421 aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagtat
      481 tgccagtcac tatggcgtgc tgcttgcgct ctatgcgttg atgcaatttc tttgcgcacc
      541 cgttctcgga gccctgtccg accgctttgg ccgccgtcca gtcctgctcg cttcgctcct
      601 tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga ttctctacgc
      661 cggacgcatc gtggcgggca tcacgggtgc cacaggtgcg gttgctggtg cctatatcgc
      721 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      781 cgtgggtatg gtggtaggcc ccgtggccgg gggactgttg ggtgccatct ccttgcatgc
      841 accattcctt gcggcggcgg tgctcaacgg cctcaacctc ctcctgggct gcttccttat
      901 gcaggaatcg cataagggag agcgccgtcc gatgcccttg cgtgccttca atccagtcag
      961 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg ttttctttat
     1021 catgcaactc gtaggacagg ttccggcagc gctctgggtc attttcggcg aggaccgctt
     1081 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     1141 cgctcaagcc ttcgtcacgg gccccgccac caaacgtttc ggcgagaagc aggccattat
     1201 cgcgggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgcggctg
     1261 gatggccttc cccattatga ttcttctcgc ttccggcggc atcggtatgc ccgcgttgca
     1321 ggccatgctg tcccgccaag tagatgacga ccatcaggga cagcttcaag ggtcgctcgc
     1381 ggctcttacc agcctcactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     1441 ctcggcgagc acatggaacg ggttggcatg gattgtaggt gccgcccttt accttgtctg
     1501 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctaat aatactagct
     1561 ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
     1621 tacttcgcgt ttgccacctg acgtctaaga aaaggaatat tcagcaattt gcccgtgccg
     1681 aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gctacgtaac aatacgcaaa
     1741 ccgcctctcg ttccttatca tctggcgaat cggacccaca agagcactgt ttacagctag
     1801 ctcagtccta ggtattatgc tagctactag agaaagagga gaaatactag atggcttcct
     1861 ccgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt tccgttaacg
     1921 gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt acccagaccg
     1981 ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc ctgtccccgc
     2041 agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg gactacctga
     2101 aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa gacggtggtg
     2161 ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac aaagttaaac
     2221 tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc atgggttggg
     2281 aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa atcaaaatgc
     2341 gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc tacatggcta
     2401 aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac atcacctccc
     2461 acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt cactccaccg
     2521 gtgcttaata acgctgatag tgctagtgta gatcgctact agagccaggc atcaaataaa
     2581 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2641 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt
     2701 agcggccgct gcaggagtca ctaagggtta gttagttaga ttagcagaaa gtcaaaagcc
     2761 tccgaccgga ggcttttgac taaaacttcc cttggggtta tcattggggc tcactcaaag
     2821 gcggtaatca gatgcggcga gcggtatcag ctcactcaaa ggcggtaatc agataaaaaa
     2881 aatccttagc tttcgctaag gatgatttct gctagtacgt acggtttcga agcttgctct
     2941 tcttaagcat tactcgcatc cattctcagg ctgtctcgtc tcgtctccgc gcgcggtctt
     3001 cctggagatg atcaaaccta ggccaggata catagattac cacaactccg agcccttcca
     3061 cccgccggcg agacctctag gccgcactcg agcaccacca ccaccaccac tgagatccgg
     3121 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     3181 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3241 tatccggata aaccttgctt cttcgcggtg tttcagtggt gcgtacaatt aagggattat
     3301 ggtaaatata tgagctatgc gataacttta actgtgaagc gatgaaccca ttacaggcaa
     3361 agccaattac tcctgacagt ggtttagcca gaagcagggc taccaagacc aatgcaataa
     3421 gtaatatatc gttttgctat cgtgcatcgc tcgcgctcag ttccattgtg cttttttaag
     3481 ctgtcgtttt tcttacggta tataccggtt ttttatggcg tggtttctta acttgttcag
     3541 ctactgatgg acccattgta tctaggtagt caactagctt tgttagatca taaatattgc
     3601 gaccaccata tcggcgatca ctcttcgatg ttggtttctt gtccaagaga ttagcttttt
     3661 caagatcatt gatagctctc tgaacagtcc gtacagaaac ccccatacgt atggctagac
     3721 tttccattga cggatgcggc cactcttgca aactccacca gtgaacgatc aggttaagta
     3781 gtgtgttaaa ggccactgaa gttagctttt tctcgttttg tataaaaaac aatacggtag
     3841 gcactgctgt ccagccaaga gacaaaccgc cagctttcca tttattctta acggagtaag
     3901 tcattgattt tcctaagccc caaaatattt aaagtatata ttatatggta tattcatatg
     3961 aatagggtga cactggcgcc attattgtgc aaccaaaaaa gactactctg aaaacgagga
     4021 aaagattttt tcctgcctga attagatacg gagttagcga tatgaaaacc gaacaacgtc
     4081 atgatcttgt taaagatatt gaggtttttg gcgtatcctt gtctctgttg atttccagag
     4141 cgaatgagaa gtctgttaca atgccatctg gtctaagtcg ggagcagaga agagcatggg
     4201 cagcggagca ggcgcgcaaa atccacaatt gaatattgtc tcattctctg agaccttcaa
     4261 cctttattac acatccagat attctgcaaa aacactcgat aaaatcgatg atttcattga
     4321 gcattttgaa aaatacaatc tctttggcga tcctttaaaa ggatatccag cttggactgg
     4381 caaagtatcg ccatcgtgga aagtgcctga tcattacgaa aacaaagaag ctattgagaa
     4441 gtatgctaga gctaacaaat tatggcatgc tcatttaggc gatccggttt ttaaagatac
     4501 gtttcatggg aaatacaagg tttctgactg ggttattcat ttccagcggc tgacaccgaa
     4561 ccatataaag cttttagagc ttggatacca tgacccaatg aagctacctc aaaactttga
     4621 atgatcgagc ggcaggctaa atgaaatctt gagattcatt cagtctcgtc gtaatctcac
     4681 tattgtaaaa acgaaaaaac caccctggca ggtggttttt cgaaggttag ttaatcctgg
     4741 cagattctct aaccgtggta acagtcttgt gcgagacatg tcaccaaata ctgtcctttc
     4801 agtgtagcct cagttaggcc gccacttcaa gaactctcgt tacatctctc gcacatcctg
     4861 cttaccagtg gccgttgcca gtggcgttaa gtcgtgtctt accgggttgg actcaagacg
     4921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4981 cttggagcga acgacctaca ccgaaccgag ataccaacag cgtgagctat gagaaagcgc
     5041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5221 gaaaaacgcc tcccgcgcgc ccttcttctg ggtctttgtc ttttgctcac atgttctttc
     5281 cggttttatc ccccgattct gtggataacc gtattaccgc ctttgagtga gctgacaccg
     5341 ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaagagcgcc
     5401 ttatgtgaca ttttctcctt acgctcctct atgccgttct gcatcctgtc cggatgcgtt
     5461 atatcccggt aagattttcc gcttcaaagc gtgtctgtat gctgttctgg agttcttctg
     5521 cgagttcgtg cagtttctca cacatggcgg cctgttcgtc ggcattgagt gcgtccagtt
     5581 tttcgagcag cgtcaggctc tgacttttta tgaatcccgc catgttgagt acggcttgct
     5641 gctgcttatt catcttttcg ttttctccgt tctgtctgtc atctgcgttg tgtgattata
     5701 tcgcgtacca cttttcgact gttttgctgc cgctattctg ccgcttggct ttttgacggg
     5761 catttctgtc agacaacact gtcactgcca aaaaactgcc gtgcctttgt cggtaattcg
     5821 agcttgctga caggacagga tgtgcaattg ttataccgcg catacatgca cgctattaca
     5881 attgccctgg tcagggcttt gccccgacac ccatgtcaga tacggagcca tgttttatga
     5941 caaaacgaag tggaagtaat acgcgcaggc gggctatcag tcgccctgtt cgtctgacgg
     6001 cagaagaaga ccaggaaatc agaaaaaggg ctgctgaatg cggcaagacc gtttccggtt
     6061 ttttacgggc ggcagctctc ggtaagaaag ttaactcact gactgatgat cgggtactga
     6121 aagaagttat gagactgggg gcgttacaga aaaaactctt tatcgacggc aagcgtgtcg
     6181 gggacaggga gtatgcggag gtgctgatcg ctattacgga gtatcaccgt gccctgttat
     6241 ccaggcttat ggcagattag cttcccggag agaaaactgt cgaaaactga cggtatgaac
     6301 accgtaagct cccaaagtga tcaccattcg ctttcatgca tagctatgca gcgagctgaa
     6361 aacgatcctg acgcatcctt cctgtttttc cggggtaaaa catctctttt tgcggtgtct
     6421 gcgtcagaat cgcgttcagc gc
//
