LOCUS       sequence-331396-        9938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli Nissle pMUT1-derived plasmid with ampicillin resistance and 
            temperature sensitive curli expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9938)
  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 9938)
  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: pM1s3ATScsg-Etag
FEATURES             Location/Qualifiers
     source          1..9938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(232..261)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    308..329
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(409..477,478..1269)
                     /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(1399..1428)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_RNA        1669..1718
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        5101..5178
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6334..6378
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      6430..6477
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             6649..6666
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6733..6780
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     8681..8700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8923..8940
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgtttttccg gggtaaaaca tctctttttg cggtgtctgc gtcagaatcg cgttcagcgc
       61 ccctaaatct agaatgcggt cgagctcgta aacttggtct gacagctcta gctccggcaa
      121 aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
      181 cgtttgccac ctgacgtcta agaaaaggaa tattcagcaa tttgcccgtg ccgaagaaag
      241 gcccacccgt gaaggtgagc cagtgagttg attgctacgt aattagttag ttagccctta
      301 gtgactcgaa ttcgcggccg cttctagagc taaatacatt caaatatcta tccgctcatg
      361 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagtattcaa
      421 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      481 ccagaaacgc tggtgaaagt aaaagatgcc gaagatcagt tgggtgcacg tgtgggttac
      541 atcgaactgg acctcaacag cggtaagatt cttgagagtt ttcgccccga agaacgtttc
      601 ccaatgatga gcacttttaa agttctgctc tgtggcgcgg tattatcccg tattgacgcc
      661 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      721 ccagtcacag aaaagcatct tacggacggc atgacagtac gcgaattatg cagcgctgcc
      781 ataaccatga gtgataacac ggcggccaac ttacttctga caacgatcgg aggaccgaag
      841 gagcttaccg cttttttgca caacatgggt gatcatgtaa ctcgccttga tcgttgggaa
      901 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagctatg
      961 gcaacaacgt tgcgcaaact cttaactggc gaacttctta ctctcgcttc ccggcaacaa
     1021 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1081 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtcccg cggtattatt
     1141 gcagccctgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagc
     1201 caggcaacta tggacgaacg taatcgccag atcgctgaga taggtgcctc cctgattaag
     1261 cattggtaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt
     1321 ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga aaaggaatat
     1381 tcagcaattt gcccgtgccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt
     1441 gctacgtaac aatacgcaaa ccgcctctcg ttccttatca tctggcgaat cggacccaca
     1501 agagcactgc attactcgca tccattctca ggctgtctcg tctcgtctcc tgacctcctg
     1561 ccagcaatag taagacaaca cgcaaagtcc gatctcgatc ccgcgaaatt ttaatttgtt
     1621 tgttagttag tttatttgtt ggtttgtttg tgttataata tcctctagct gtcaccggat
     1681 gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaatt ttgtttaagt
     1741 acgaccaggt ccagggtgac aacatgaaaa acaaattgtt atttatgatg ttaacaatac
     1801 tgggtgcgcc tgggattgca gccgcagcag gttatgattt agctaattca gaatataact
     1861 tcgcggtaaa tgaattgagt aagtcttcat ttaatcaggc agccataatt ggtcaagctg
     1921 ggactaataa tagtgctcag ttacggcagg gaggctcaaa acttttggcg gttgttgcgc
     1981 aagaaggtag tagcaaccgg gcaaagattg accagacagg agattataac cttgcatata
     2041 ttgatcaggc gggcagtgcc aacgatgcca gtatttcgca aggtgcttat ggtaatactg
     2101 cgatgattat ccagaaaggt tctggtaata aagcaaatat tacacagtat ggtactcaaa
     2161 aaacggcaat tgtagtgcag agacagtcgc aaatggctat tcgcgtgaca caacgttaat
     2221 ttccattcga cttttaaatc aatccgatgg gggttttaca tgaaactttt aaaagtagca
     2281 gcaattgcag caatcgtatt ctccggtagc gctctggcag gtgttgttcc tcagtacggc
     2341 ggcggcggta accacggtgg tggcggtaat aatagcggcc caaattctga gctgaacatt
     2401 taccagtacg gtggcggtaa ctctgcactt gctctgcaaa ctgatgcccg taactctgac
     2461 ttgactatta cccagcatgg cggcggtaat ggtgcagatg ttggtcaggg ctcagatgac
     2521 agctcaatcg atctgaccca acgtggcttc ggtaacagcg ctactcttga tcagtggaac
     2581 ggcaaaaatt ctgaaatgac ggttaaacag ttcggtggtg gcaacggtgc tgcagttgac
     2641 cagactgcat ctaactcctc cgtcaacgtg actcaggttg gctttggtaa caacgcgacc
     2701 gctcatcagt acggtggatc tggtagcagc ggctctggtg gttctggggg tgcgccggtt
     2761 ccgtacccgg acccgctgga accgcgtggt ggctcgggtt catctggtag tggcggttcg
     2821 ggttaataca tcatttgtat tacagaaaca gggcgcaagc cctgtttttt ttcgggagaa
     2881 gaatatgaat acgttattac tccttgcggc actttccagt cagataacct ttaatacgac
     2941 ccagcaaggg gatgtgtata ccattattcc tgaagtcact cttactcaat cttgtctgtg
     3001 cagagtacaa atattgtccc tgcgcgaagg cagttcaggg caaagtcaga cgaagcaaga
     3061 aaagaccctt tcattgcctg ctaatcaacc aattgctttg acgaagttga gtttaaatat
     3121 ttccccggac gatcgggtga aaatagttgt tactgtttct gatggacagt cacttcattt
     3181 atcacaacaa tggccgccct cttcagaaaa gtcttaaggt ttcctgggca aacgataacc
     3241 tcaggcgata aagccatgaa acgttattta cgctggattg tggcggcaga atttctgttc
     3301 gccgcaggga atcttcacgc cgttgaggta gaagtcccgg gattgctaac tgaccatact
     3361 gtttcatcta ttggccatga tttttaccga gcctttagtg ataaatggga aagtgactat
     3421 acgggtaact taacgattaa tgaaaggccc agtgcacgat ggggaagctg gatcactata
     3481 acggtcaatc aggacgttat tttccagact tttttatttc cgttgaaaag agacttcgag
     3541 aaaactgtcg tctttgcact gattcaaact gaagaagcac taaatcgtcg ccagataaat
     3601 caggcgttat taagtacggg cgatttggcg catgatgaat tctaaataaa aaattgttcg
     3661 gaggctgcaa tgcgtgtcaa acatgcagta gttctactca tgcttatttc gccattaagt
     3721 tgggctggaa ccatgacttt ccagttccgt aatccaaact ttggtggtaa cccaaataat
     3781 ggcgcttttt tattaaatag cgctcaggcc caaaactctt ataaagatcc gagctataac
     3841 gatgactttg gtattgaaac accctcagcg ttagataact ttactcaggc catccagtca
     3901 caaattttag gtgggctact gtcgaatatt aataccggta aaccgggccg catggtgacc
     3961 aacgattata ttgtcgatat tgccaaccgc gatggtcaat tgcagttgaa cgtgacagat
     4021 cgtaaaaccg gacaaacctc gaccatccag gtttcgggtt tacaaaataa ctcaaccgat
     4081 ttttaagccc cagcttcata aggaaaataa tcatgcagcg cttatttctt ttggttgccg
     4141 tcatgttact gagcggatgc ttaaccgccc cgcctaaaga agccgccaga ccgacattaa
     4201 tgcctcgtgc tcagagctac aaagatttga cccatctgcc agcgccgacg ggtaaaatct
     4261 ttgtttcggt atacaacatt caggacgaaa ccgggcaatt taaaccctac ccggcaagta
     4321 acttctccac tgctgttccg caaagcgcca cggcaatgct ggtcacggca ctgaaagatt
     4381 ctcgctggtt tataccgctg gagcgccagg gcttacaaaa cctgcttaac gagcgcaaga
     4441 ttattcgtgc ggcacaagaa aacggcacgg ttgccattaa taaccgaatc ccgctgcaat
     4501 ctttaacggc ggcaaatatc atggttgaag gttcgattat cggttatgaa agcaacgtca
     4561 aatctggcgg ggttggggca agatattttg gcatcggtgc cgacacgcaa taccagctcg
     4621 atcagattgc cgtgaacctg cgcgtcgtca atgtgagtac cggcgagatc ctttcttcgg
     4681 tgaacaccag taagacgata ctttcctatg aagttcaggc cggggttttc cgctttattg
     4741 actaccagcg cttgcttgaa ggggaagtgg gttacacctc gaacgaacct gttatgctgt
     4801 gcctgatgtc ggctatcgaa acaggggtca ttttcctgat taatgatggt atcgaccgtg
     4861 gtctgtggga tttgcaaaat aaagcagaac ggcagaatga cattctggtg aaataccgcc
     4921 atatgtcggt tccaccggaa tcctgataac tcggtaccaa attccagaaa agaggcctcc
     4981 cgaaaggggg gccttttttc gttttggtcc aaaaaagccc gcacctgaca gtgcgggctt
     5041 tttttttaag cttgcggccg cataatgctt aagtcgaaca gaaagtaatc gtatgagctc
     5101 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     5161 caattcaggg tggtgaatgt atgtgaggta gttatacata tgcgtccggc gacatacgaa
     5221 ccagaacaga ttattgaagc agggctggcc ctgcaggctg aaggacggaa tatcaccggg
     5281 ttcgcactac gtaaccaggt gggtggcggc aatccgacac gtctccgcca gatatgggac
     5341 gaataccagg cttcacagag cacggtcgtc actgaactcg ttgccgagct gccagtggaa
     5401 gtggctgaag aagtgaaggc cgtctccgcc gcgctgtccg aacgcatcac ccagctggcg
     5461 acagaactga atgacaaggc ggtccgggct gcagaacgcc gggttgcgga agtcacgcgt
     5521 gctgccggtg aacagaccgc acaggcagag cgggagctgg ccgacgccgc gcagacagtc
     5581 gacgacctgg aagaaaaact ggttgaactg caggacagat atgacagttt gacgctggcg
     5641 ctggagtcag aacgttcact gcgtcagcag catgatgtgg agatggccca gctgaaagag
     5701 cgtcttgcgg ccgctgaaga gaatacccgt cagcgagagg aacggtatca ggagcagagg
     5761 acagtgctgc aggatgcgct taatgcggag caggcacagc acataaacac gcgggaagac
     5821 cagcagaaac gactggagca aatttctgcc gaagctaatg cgcgtacaga agaactgaag
     5881 tctgaacgcg ataaagtcaa tactctcctt acccgccttg aatcgcagga aaatgcgctg
     5941 gcctcagaac gtcagcagca tctggccacc cgcgaaacgc tgcagcaacg cctcgagcag
     6001 gccatcgctg acacgcaggc gcgcgccggt gagattgcac ttgaacgtga cagagtcagc
     6061 agcctcaccg caaggctgga atcgcaggaa aaggcctcct cggagcaact ggtgcgtatg
     6121 ggcagtgaaa tagccagtct gacagagcgt tgcacacagc tggaaaacca gcgtgatgat
     6181 gcccgtctgg agacgatggg ggagaaagaa acggtcgcgg cactgcgtgg tgaggctgaa
     6241 gccctgaagc gtcagaacca gtcactgatg gcggcgcttt caggcaataa acagaccggt
     6301 ggccagaatg cgtgaggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa
     6361 cgccagcaca tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct
     6421 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     6481 actcggtacc aaattttcga aaaaagacgc tgaaaagcgt cttttttcgt tttggtccga
     6541 gccaactccc tttacaacct cactcaagtc cgttagagcc aggatacata gattaccaca
     6601 actccgagcc cttccacccg ccggcgagac ctctaggccg cactcgagca ccaccaccac
     6661 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     6721 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     6781 ctgaaaggag gaactatatc cggataaacc ttgcttcttc gcggtgtttc agtggtgcgt
     6841 acaattaagg gattatggta aatatatgag ctatgcgata actttaactg tgaagcgatg
     6901 aacccattac aggcaaagcc aattactcct gacagtggtt tagccagaag cagggctacc
     6961 aagaccaatg caataagtaa tatatcgttt tgctatcgtg catcgctcgc gctcagttcc
     7021 attgtgcttt tttaagctgt cgtttttctt acggtatata ccggtttttt atggcgtggt
     7081 ttcttaactt gttcagctac tgatggaccc attgtatcta ggtagtcaac tagctttgtt
     7141 agatcataaa tattgcgacc accatatcgg cgatcactct tcgatgttgg tttcttgtcc
     7201 aagagattag ctttttcaag atcattgata gctctctgaa cagtccgtac agaaaccccc
     7261 atacgtatgg ctagactttc cattgacgga tgcggccact cttgcaaact ccaccagtga
     7321 acgatcaggt taagtagtgt gttaaaggcc actgaagtta gctttttctc gttttgtata
     7381 aaaaacaata cggtaggcac tgctgtccag ccaagagaca aaccgccagc tttccattta
     7441 ttcttaacgg agtaagtcat tgattttcct aagccccaaa atatttaaag tatatattat
     7501 atggtatatt catatgaata gggtgacact ggcgccatta ttgtgcaacc aaaaaagact
     7561 actctgaaaa cgaggaaaag attttttcct gcctgaatta gatacggagt tagcgatatg
     7621 aaaaccgaac aacgtcatga tcttgttaaa gatattgagg tttttggcgt atccttgtct
     7681 ctgttgattt ccagagcgaa tgagaagtct gttacaatgc catctggtct aagtcaggag
     7741 cagagaagag catgggcagc ggagcaggcg cgcaaaatcc acaattgaat attgtctcat
     7801 tctctgagac cttcaacctt tattacacat ccagatattc tgcaaaaaca ctcgataaaa
     7861 tcgatgattt cattgagcat tttgaaaaat acaatctctt tggcgatcct ttaaaaggat
     7921 atccagcttg gactggcaaa gtatcgccat cgtggaaagt gcctgatcat tacgaaaaca
     7981 aagaagctat tgagaagtat gctagagcta acaaattatg gcatgctcat ttaggcgatc
     8041 cggtttttaa agatacgttt catgggaaat acaaggtttc tgactgggtt attcatttcc
     8101 agcggctgac accgaaccat ataaagcttt tagagcttgg ataccatgac ccaatgaagc
     8161 tacctcaaaa ctttgaatga tcgagcggca ggctaaatga aatcttgaga ttcattcagt
     8221 ctcgtcgtaa tctcactatt gtaaaaacga aaaaaccacc ctggcaggtg gtttttcgaa
     8281 ggttagttaa tcctggcaga ttctctaacc gtggtaacag tcttgtgcga gacatgtcac
     8341 caaatactgt cctttcagtg tagcctcagt taggccgcca cttcaagaac tctcgttaca
     8401 tctctcgcac atcctgctta ccagtggccg ttgccagtgg cgttaagtcg tgtcttaccg
     8461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga accgagatac caacagcgtg
     8581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8641 gcagggtcgg aacaggagag cgcacgaggg agcttccagt gggaaacgcc tggtatcttt
     8701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8761 gggggcggag cctatggaaa aacgcctccc gcgcgccctt cttctgggtc tttgtctttt
     8821 gctcacatgt tctttccggt tttatccccc gattctgtgg ataaccgtat taccgccttt
     8881 gagtgagctg acaccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
     8941 gaagcggaag agcgccttat gtgacatttt ctccttacgc tcctctatgc cgttctgcat
     9001 cctgtccgga tgcgttatat cccggtaaga ttttccgctt caaagcgtgt ctgtatgctg
     9061 ttctggagtt cttctgcgag ttcgtgcagt ttctcacaca tggcggcctg ttcgtcggca
     9121 ttgagtgcgt ccagtttttc gagcagcgtc aggctctgac tttttatgaa tcccgccatg
     9181 ttgagtacgg cttgctgctg cttattcatc ttttcgtttt ctccgttctg tctgtcatct
     9241 gcgttgtgtg attatatcgc gtaccacttt tcgactgttt tgctgccgct attctgccgc
     9301 ttggcttttt gacgggcatt tctgtcagac aacactgtca ctgccaaaaa actgccgtgc
     9361 ctttgtcggt aattcgagct tgctgacagg acaggatgtg caattgttat accgcgcata
     9421 catgcacgct attacaattg ccctggtcag ggctttgccc cgacacccat gtcagatacg
     9481 gagccatgtt ttatgacaaa acgaagtgga agtaatacgc gcaggcgggc tatcagtcgc
     9541 cctgttcgtc tgacggcaga agaagaccag gaaatcagaa aaagggctgc tgaatgcggc
     9601 aagaccgttt ccggtttttt acgggcggca gctctcggta agaaagttaa ctcactgact
     9661 gatgatcggg tactgaaaga agttatgaga ctgggggcgt tacagaaaaa actctttatc
     9721 gacggcaagc gtgtcgggga cagggagtat gcggaggtgc tgatcgctat tacggagtat
     9781 caccgtgccc tgttatccag gcttatggca gattagcttc ccggagagaa aactgtcgaa
     9841 aactgacggt atgaacaccg taagctccca aagtgatcac cattcgcttt catgcatagc
     9901 tatgcagcga gctgaaaacg atcctgacgc atccttcc
//
