LOCUS       p2H12Matry              4637 bp ds-DNA     circular SYN 14-MAY-2024
DEFINITION  E. coli plasmid with 2H12Matry  sensor. UJ11208.
ACCESSION
VERSION     .
KEYWORDS    pATT-Dest.
SOURCE      
  ORGANISM  .
REFERENCE   1  (bases 1 to 2911)
  AUTHORS   Nadler, DC, Morgan, SA, Flamholz, A, Kortright, KE, Savage and DF
  TITLE     Rapid construction of metabolite biosensors using domain-insertion
            profiling.
  JOURNAL   Nat Commun. 2016 Jul 29;7:12266. doi: 10.1038/ncomms12266.
  PUBMED    27470466
REFERENCE   2  (bases 1 to 2911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     promoter        <7..108
                     /created_by="User"
                     /label="Ptet"
     source          <7..>2832
                     /organism="Escherichia coli str. K-12 substr. MG1655"
                     /mol_type="genomic DNA"
                     /strain="K-12"
                     /sub_strain="MG1655"
                     /db_xref="taxon:511145"
     regulatory      25..43
                     /created_by="User"
                     /modified_by="User"
                     /label="TetO"
     -35             44..49
                     /created_by="User"
     regulatory      50..68
                     /created_by="User"
                     /label="TetO"
     -10             67..72
                     /created_by="User"
     RBS             109..123
                     /created_by="User"
     gene            124..747
                     /created_by="User"
                     /label="tetR gene"
     CDS             124..747
                     /created_by="User"
                     /label="tetR CDS"
     primer_bind     713..743
                     /Sequence="TAGAAAAACAACTTAAATGTGAAAGTGGGTC"
                     /Extension="ACTAGT"
                     /created_by="kaczmarc"
                     /label=13378
     3x_stop_codon   748..756
                     /created_by="User"
                     /label="3x stop"
     RBS             763..771
                     /created_by="User"
     CDS             778..2820
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="cdGreen-Matry"
     ORF             778..1047
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD CTD 1"
     misc_feature    1048..1068
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="2H12_N-linker"
     ORF             1069..1341
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="cpGFP_original Cterm"
     misc_feature    1231
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     ORF             1375..2070
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="mScarlet-I"
     ORF             2107..>2541
                     /created_by="kaczmarc"
                     /label="cpGFP_original Nterm"
     misc_feature    2542..2556
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="2H12_C-linker"
     ORF             2557..2820
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD CTD 2"
     misc_feature    2826^2827
                     /Original_Bases="AGGAGGTTAATATGAGCAAAGGTGAAGCCGTGATCAAAGAA
                     TTTATGCGCTTTAAAGTTCACATGGAAGGTAGCATGAATGGCCACGAATTTGAAATTG
                     AAGGTGAAGGCGAAGGTCGTCCGTATGAAGGCACCCAGACCGCAAAACTGAAAGTTAC
                     CAAAGGTGGTCCGCTGCCGTTTAGCTGGGATATTCTGAGTCCGCAGTTTATGTATGGT
                     AGCCGTGCATTTATCAAACATCCGGCAGATATTCCGGATTACTATAAACAGAGCTTTC
                     CGGAAGGTTTTAAATGGGAACGTGTGATGAATTTTGAAGATGGTGGTGCAGTTACCGT
                     TACACAGGATACCAGCCTGGAAGATGGCACCCTGATCTATAAAGTTAAACTGCGTGGC
                     ACCAATTTTCCGCCTGATGGTCCGGTTATGCAGAAAAAAACCATGGGTTGGGAAGCAA
                     GCACCGAACGTCTGTATCCTGAAGATGGCGTTCTGAAAGGTGATATCAAAATGGCACT
                     GCGTCTGAAAGATGGCGGTCGTTATYTGGCAGATTTCAAAACCACCTACAAAGCCAAA
                     AAACCGGTTCAGATGCCTGGTGCATATAATGTTGATCGCAAACTGGATATTACCAGCC
                     ACAATGAAGATTATACCGTGGTGGAACAGTATGAACGTAGCGAAGGCCGTCATAGTAC
                     CGGTGGTATGGATGAACTGTACAAGTGAGCTAGC"
                     /label="AGGAGGTTAATATGAGCAAAGGTGAAGCCGTGATCAAAGAATTTATGCGC
                     TTTAAAGTTCACATGGAAGGTAGCATGAATGGCCACGAATTTGAAATTGAAGGTGAAG
                     GCGAAGGTCGTCCGTATGAAGGCACCCAGACCGCAAAACTGAAAGTTACCAAAGGTGG
                     TCCGCTGCCGTTTAGCTGGGATATTCTGAGTCCGCAGTTTATGTATGGTAGCCGTGCA
                     TTTATCAAACATCCGGCAGATATTCCGGATTACTATAAACAGAGCTTTCCGGAAGGTT
                     TTAAATGGGAACGTGTGATGAATTTTGAAGATGGTGGTGCAGTTACCGTTACACAGGA
                     TACCAGCCTGGAAGATGGCACCCTGATCTATAAAGTTAAACTGCGTGGCACCAATTTT
                     CCGCCTGATGGTCCGGTTATGCAGAAAAAAACCATGGGTTGGGAAGCAAGCACCGAAC
                     GTCTGTATCCTGAAGATGGCGTTCTGAAAGGTGATATCAAAATGGCACTGCGTCTGAA
                     AGATGGCGGTCGTTATYTGGCAGATTTCAAAACCACCTACAAAGCCAAAAAACCGGTT
                     CAGATGCCTGGTGCATATAATGTTGATCGCAAACTGGATATTACCAGCCACAATGAAG
                     ATTATACCGTGGTGGAACAGTATGAACGTAGCGAAGGCCGTCATAGTACCGGTGGTAT
                     GGATGAACTGTACAAGTGAGCTAGC"
                     /note="Geneious type: Editing History Deletion"
     source          <2833..>4637
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2858..2962
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             2963..3823
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     primer_bind     complement(3181..3200)
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      3994..4582
                     /direction="RIGHT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     4483..4502
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     misc_feature    4637^4638
                     /Original_Bases="ACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAAC
                     CGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCA
                     GCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCATCAGCAACCGCACCTGTGGCGCCG
                     GTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGATCCCGCGAAATT
                     AATACGACTCACTATAGGGACACCACAACGGTTTCCCTCTAGAAATAATTTTGTTTAA
                     CTTTAAGAAGGAGATATACCATCGTCTCTGTCCAGAGACCATTAATGCAGCTGGCACG
                     ACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCT
                     CACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGA
                     ATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGC
                     TTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCACTGG
                     CCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCT
                     TGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGC
                     CCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCC
                     TTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTC
                     TGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGA
                     CGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGGTCTCTAGCGAGAGACGG
                     TTAAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCT
                     GAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGC
                     TGAAAGGAGGAACTATATCCGGATATCCACAGGACGGGTGTGGTCGCCATGATCGCGT
                     AGT"
                     /label="ACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACC
                     GCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAG
                     TGAGCGAGGAAGCGGAAGAGCGCCATCAGCAACCGCACCTGTGGCGCCGGTGATGCCG
                     GCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGATCCCGCGAAATTAATACGACT
                     CACTATAGGGACACCACAACGGTTTCCCTCTAGAAATAATTTTGTTTAACTTTAAGAA
                     GGAGATATACCATCGTCTCTGTCCAGAGACCATTAATGCAGCTGGCACGACAGGTTTC
                     CCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTA
                     GGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGC
                     GGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTGCATGCC
                     TGCAGGTCGACTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTT
                     TACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACA
                     TCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAA
                     CAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATC
                     TGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGC
                     ATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGT
                     CTGCTCCCGGCATCCGCTTACAGACAAGCTGGTCTCTAGCGAGAGACGGTTAAGATCC
                     GGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAA
                     CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAG
                     GAACTATATCCGGATATCCACAGGACGGGTGTGGTCGCCATGATCGCGTAGT"
                     /note="Geneious type: Editing History Deletion"
ORIGIN      
        1 acatgttttc gtcttcacct cgagtcccta tcagtgatag agattgacat ccctatcagt
       61 gatagagata ctgagcacat cagcaggacg cactgaccga atttattaag gaggaggtga
      121 tccatgtcta gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
      181 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
      241 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
      301 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
      361 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
      421 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
      481 ttatgccaac aaggtttttc actagagaat gcattatatg cgctcagcgc tgtggggcat
      541 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
      601 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
      661 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
      721 caacttaaat gtgaaagtgg gtcttaataa taataattcc gaaggaggag gtgatccatg
      781 aattcagagc cgccgccgaa gctcgtcctt gacctggagc gcctcgcgca cgtcccccag
      841 gagaaggcgg gcccgctcca gcgctacgcg gcgacgatcc agtcgcagcg cggcgactac
      901 aacggcaagg tgctgtcgat ccgtcaggac gacctgcgca ccctggccgt gatctacgac
      961 cagtcgccct cggtcctcac cgagcagctg atcagctggg gcgtcctcga cgcggacgcg
     1021 cgccgcgccg tcgcccacga ggagaacccg aagaatatgc cggtcccgta taacgtcttt
     1081 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     1141 gaggacggcg gcgtgcagct cgcctatcac taccagcaga acacccccat cggcgacggc
     1201 cccgtgctgc tgcccgacaa ccactacctg ggcgtgcagt ccaaactgag caaagacccc
     1261 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1321 ggcatggacg agctgtacaa gggcggtacc ggaggcagcg gcggcacggg cggaagcaag
     1381 aaatatggtg aagccgtgat caaagaattt atgcgcttta aagttcacat ggaaggtagc
     1441 atgaatggcc acgaatttga aattgaaggt gaaggcgaag gtcgtccgta tgaaggcacc
     1501 cagaccgcaa aactgaaagt taccaaaggt ggtccgctgc cgtttagctg ggatattctg
     1561 agtccgcagt ttatgtatgg tagccgtgca tttatcaaac atccggcaga tattccggat
     1621 tactataaac agagctttcc ggaaggtttt aaatgggaac gtgtgatgaa ttttgaagat
     1681 ggtggtgcag ttaccgttac acaggatacc agcctggaag atggcaccct gatctataaa
     1741 gttaaactgc gtggcaccaa ttttccgcct gatggtccgg ttatgcagaa aaaaaccatg
     1801 ggttgggaag caagcaccga acgtctgtat cctgaagatg gcgttctgaa aggtgatatc
     1861 aaaatggcac tgcgtctgaa agatggcggt cgttatctgg cagatttcaa aaccacctac
     1921 aaagccaaaa aaccggttca gatgcctggt gcatataatg ttgatcgcaa actggatatt
     1981 accagccaca atgaagatta taccgtggtg gaacagtatg aacgtagcga aggccgtcat
     2041 agtaccggtg gtatggatga actgtacaag ggcgggagcg gaggcacggg aggtaccgga
     2101 gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2161 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2221 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2281 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2341 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacattca ggagcgcacc
     2401 atcttcttca aggacgacgg caactataag acacgcgctg aggttaagtt cgagggcgac
     2461 actctggtta accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2521 ggccataagc ttgaatataa catgttgtcg gagccggagc cgccgccgaa gctcgtcctt
     2581 gacctggagc gcctcgcgca cgtcccccag gagaaggcgg gcccgctcca gcgctacgcg
     2641 gcgacgatcc agtcgcagcg cggcgactac aacggcaagg tgctgtcgat ccgtcaggac
     2701 gacctgcgca ccctggccgt gatctacgac cagtcgccct cggtcctcac cgagcagctg
     2761 atcagctggg gcgtcctcga cgcggacgcg cgccgcgccg tcgcccacga ggagaactga
     2821 actagtgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2881 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2941 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3001 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3061 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3121 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3181 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     3241 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3301 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3361 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3421 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3481 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     3541 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3601 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3661 tggagccggt gagcgtggct ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3721 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3781 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3841 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3901 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3961 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4021 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4081 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4141 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4201 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4261 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4321 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4381 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4441 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4501 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4561 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4621 ttgctggcct tttgctc
//
