LOCUS       sequence-322876-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli Nissle pMUT1-derived plasmid with ampicillin resistance and 
            constitutive GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  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 6018)
  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: pM1s3AsG
FEATURES             Location/Qualifiers
     source          1..6018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(186..215)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    262..283
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(363..431,432..1223)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      complement(1353..1382)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        1464..1498
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1507..1518
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1525..2238
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2286..2357
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2373..2400
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2683..2700
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2753..2771)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2767..2814
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4715..4734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4957..4974
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgcgttca gcgcccctaa atctagaatg cggtcgagct cgtaaacttg gtctgacagc
       61 tctagctccg gcaaaaaaac gggcaaggtg tcaccaccct gccctttttc tttaaaaccg
      121 aaaagattac ttcgcgtttg ccacctgacg tctaagaaaa ggaatattca gcaatttgcc
      181 cgtgccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattgct acgtaattag
      241 ttagttagcc cttagtgact cgaattcgcg gccgcttcta gagctaaata cattcaaata
      301 tctatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      361 atatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      421 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgccgaagat cagttgggtg
      481 cacgtgtggg ttacatcgaa ctggacctca acagcggtaa gattcttgag agttttcgcc
      541 ccgaagaacg tttcccaatg atgagcactt ttaaagttct gctctgtggc gcggtattat
      601 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      661 tggttgagta ctcaccagtc acagaaaagc atcttacgga cggcatgaca gtacgcgaat
      721 tatgcagcgc tgccataacc atgagtgata acacggcggc caacttactt ctgacaacga
      781 tcggaggacc gaaggagctt accgcttttt tgcacaacat gggtgatcat gtaactcgcc
      841 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      901 tgcctgtagc tatggcaaca acgttgcgca aactcttaac tggcgaactt cttactctcg
      961 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1021 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1081 cccgcggtat tattgcagcc ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1141 acacgacggg gagccaggca actatggacg aacgtaatcg ccagatcgct gagataggtg
     1201 cctccctgat taagcattgg taataatact agctccggca aaaaaacggg caaggtgtca
     1261 ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgtttgcca cctgacgtct
     1321 aagaaaagga atattcagca atttgcccgt gccgaagaaa ggcccacccg tgaaggtgag
     1381 ccagtgagtt gattgctacg taacaatacg caaaccgcct ctcgttcctt atcatctggc
     1441 gaatcggacc cacaagagca ctgtttacag ctagctcagt cctaggtatt atgctagcta
     1501 ctagagaaag aggagaaata ctagatgcgt aaaggcgaag agctgttcac tggtgtcgtc
     1561 cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag
     1621 ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa
     1681 ctgccggtac cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct
     1741 cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat
     1801 gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg
     1861 aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagaa
     1921 gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc
     1981 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag
     2041 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct
     2101 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac
     2161 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt
     2221 atggatgaac tgtacaaatg ataacgctga tagtgctagt gtagatcgct actagagcca
     2281 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2341 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     2401 tttatatact agtagcggcc gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc
     2461 agataaaaaa aatccttagc tttcgctaag gatgatttct gctagtacgt acggtttcga
     2521 agcttgctct tcttaagcat tactcgcatc cattctcagg ctgtctcgtc tcgtctccgc
     2581 gcgcggtctt cctggagatg atcaaaccta ggccaggata catagattac cacaactccg
     2641 agcccttcca cccgccggcg agacctctag gccgcactcg agcaccacca ccaccaccac
     2701 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     2761 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     2821 ggaggaacta tatccggata aaccttgctt cttcgcggtg tttcagtggt gcgtacaatt
     2881 aagggattat ggtaaatata tgagctatgc gataacttta actgtgaagc gatgaaccca
     2941 ttacaggcaa agccaattac tcctgacagt ggtttagcca gaagcagggc taccaagacc
     3001 aatgcaataa gtaatatatc gttttgctat cgtgcatcgc tcgcgctcag ttccattgtg
     3061 cttttttaag ctgtcgtttt tcttacggta tataccggtt ttttatggcg tggtttctta
     3121 acttgttcag ctactgatgg acccattgta tctaggtagt caactagctt tgttagatca
     3181 taaatattgc gaccaccata tcggcgatca ctcttcgatg ttggtttctt gtccaagaga
     3241 ttagcttttt caagatcatt gatagctctc tgaacagtcc gtacagaaac ccccatacgt
     3301 atggctagac tttccattga cggatgcggc cactcttgca aactccacca gtgaacgatc
     3361 aggttaagta gtgtgttaaa ggccactgaa gttagctttt tctcgttttg tataaaaaac
     3421 aatacggtag gcactgctgt ccagccaaga gacaaaccgc cagctttcca tttattctta
     3481 acggagtaag tcattgattt tcctaagccc caaaatattt aaagtatata ttatatggta
     3541 tattcatatg aatagggtga cactggcgcc attattgtgc aaccaaaaaa gactactctg
     3601 aaaacgagga aaagattttt tcctgcctga attagatacg gagttagcga tatgaaaacc
     3661 gaacaacgtc atgatcttgt taaagatatt gaggtttttg gcgtatcctt gtctctgttg
     3721 atttccagag cgaatgagaa gtctgttaca atgccatctg gtctaagtcg ggagcagaga
     3781 agagcatggg cagcggagca ggcgcgcaaa atccacaatt gaatattgtc tcattctctg
     3841 agaccttcaa cctttattac acatccagat attctgcaaa aacactcgat aaaatcgatg
     3901 atttcattga gcattttgaa aaatacaatc tctttggcga tcctttaaaa ggatatccag
     3961 cttggactgg caaagtatcg ccatcgtgga aagtgcctga tcattacgaa aacaaagaag
     4021 ctattgagaa gtatgctaga gctaacaaat tatggcatgc tcatttaggc gatccggttt
     4081 ttaaagatac gtttcatggg aaatacaagg tttctgactg ggttattcat ttccagcggc
     4141 tgacaccgaa ccatataaag cttttagagc ttggatacca tgacccaatg aagctacctc
     4201 aaaactttga atgatcgagc ggcaggctaa atgaaatctt gagattcatt cagtctcgtc
     4261 gtaatctcac tattgtaaaa acgaaaaaac caccctggca ggtggttttt cgaaggttag
     4321 ttaatcctgg cagattctct aaccgtggta acagtcttgt gcgagacatg tcaccaaata
     4381 ctgtcctttc agtgtagcct cagttaggcc gccacttcaa gaactctcgt tacatctctc
     4441 gcacatcctg cttaccagtg gccgttgcca gtggcgttaa gtcgtgtctt accgggttgg
     4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4561 cacagcccag cttggagcga acgacctaca ccgaaccgag ataccaacag cgtgagctat
     4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4801 ggagcctatg gaaaaacgcc tcccgcgcgc ccttcttctg ggtctttgtc ttttgctcac
     4861 atgttctttc cggttttatc ccccgattct gtggataacc gtattaccgc ctttgagtga
     4921 gctgacaccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     4981 gaagagcgcc ttatgtgaca ttttctcctt acgctcctct atgccgttct gcatcctgtc
     5041 cggatgcgtt atatcccggt aagattttcc gcttcaaagc gtgtctgtat gctgttctgg
     5101 agttcttctg cgagttcgtg cagtttctca cacatggcgg cctgttcgtc ggcattgagt
     5161 gcgtccagtt tttcgagcag cgtcaggctc tgacttttta tgaatcccgc catgttgagt
     5221 acggcttgct gctgcttatt catcttttcg ttttctccgt tctgtctgtc atctgcgttg
     5281 tgtgattata tcgcgtacca cttttcgact gttttgctgc cgctattctg ccgcttggct
     5341 ttttgacggg catttctgtc agacaacact gtcactgcca aaaaactgcc gtgcctttgt
     5401 cggtaattcg agcttgctga caggacagga tgtgcaattg ttataccgcg catacatgca
     5461 cgctattaca attgccctgg tcagggcttt gccccgacac ccatgtcaga tacggagcca
     5521 tgttttatga caaaacgaag tggaagtaat acgcgcaggc gggctatcag tcgccctgtt
     5581 cgtctgacgg cagaagaaga ccaggaaatc agaaaaaggg ctgctgaatg cggcaagacc
     5641 gtttccggtt ttttacgggc ggcagctctc ggtaagaaag ttaactcact gactgatgat
     5701 cgggtactga aagaagttat gagactgggg gcgttacaga aaaaactctt tatcgacggc
     5761 aagcgtgtcg gggacaggga gtatgcggag gtgctgatcg ctattacgga gtatcaccgt
     5821 gccctgttat ccaggcttat ggcagattag cttcccggag agaaaactgt cgaaaactga
     5881 cggtatgaac accgtaagct cccaaagtga tcaccattcg ctttcatgca tagctatgca
     5941 gcgagctgaa aacgatcctg acgcatcctt cctgtttttc cggggtaaaa catctctttt
     6001 tgcggtgtct gcgtcaga
//
