LOCUS       p2H12                   3890 bp ds-DNA     circular SYN 13-MAY-2024
DEFINITION  E. coli plasmid with 2H12 cdG sensor. UJ11204.
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
     trimmed         1..2
                     /Method="Max ambiguities: 2, Min 5' trim: 2, Min 3' trim:
                     2"
                     /label="Trimmed"
     promoter        <7..108
                     /created_by="User"
                     /label="Ptet"
     source          <7..>2085
                     /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..2073
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD2H12 sensor"
     ORF             778..1047
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD CTD 1"
     misc_feature    925..930
                     /created_by="kaczmarc"
                     /label="mutate"
     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"
     misc_feature    1350^1351
                     /Original_Bases="GGAGGCAGCGGCGGCACGGGCGGAAGCAAGAAATATGGTGA
                     AGCCGTGATCAAAGAATTTATGCGCTTTAAAGTTCACATGGAAGGTAGCATGAATGGC
                     CACGAATTTGAAATTGAAGGTGAAGGCGAAGGTCGTCCGTATGAAGGCACCCAGACCG
                     CAAAACTGAAAGTTACCAAAGGTGGTCCGCTGCCGTTTAGCTGGGATATTCTGAGTCC
                     GCAGTTTATGTATGGTAGCCGTGCATTTATCAAACATCCGGCAGATATTCCGGATTAC
                     TATAAACAGAGCTTTCCGGAAGGTTTTAAATGGGAACGTGTGATGAATTTTGAAGATG
                     GTGGTGCAGTTACCGTTACACAGGATACCAGCCTGGAAGATGGCACCCTGATCTATAA
                     AGTTAAACTGCGTGGCACCAATTTTCCGCCTGATGGTCCGGTTATGCAGAAAAAAACC
                     ATGGGTTGGGAAGCAAGCACCGAACGTCTGTATCCTGAAGATGGCGTTCTGAAAGGTG
                     ATATCAAAATGGCACTGCGTCTGAAAGATGGCGGTCGTTATCTGGCAGATTTCAAAAC
                     CACCTACAAAGCCAAAAAACCGGTTCAGATGCCTGGTGCATATAATGTTGATCGCAAA
                     CTGGATATTACCAGCCACAATGAAGATTATACCGTGGTGGAACAGTATGAACGTAGCG
                     AAGGCCGTCATAGTACCGGTGGTATGGATGAACTGTACAAGGGCGGGAGCGGAGGCAC
                     GGGAGGTACC"
                     /label="GGAGGCAGCGGCGGCACGGGCGGAAGCAAGAAATATGGTGAAGCCGTGAT
                     CAAAGAATTTATGCGCTTTAAAGTTCACATGGAAGGTAGCATGAATGGCCACGAATTT
                     GAAATTGAAGGTGAAGGCGAAGGTCGTCCGTATGAAGGCACCCAGACCGCAAAACTGA
                     AAGTTACCAAAGGTGGTCCGCTGCCGTTTAGCTGGGATATTCTGAGTCCGCAGTTTAT
                     GTATGGTAGCCGTGCATTTATCAAACATCCGGCAGATATTCCGGATTACTATAAACAG
                     AGCTTTCCGGAAGGTTTTAAATGGGAACGTGTGATGAATTTTGAAGATGGTGGTGCAG
                     TTACCGTTACACAGGATACCAGCCTGGAAGATGGCACCCTGATCTATAAAGTTAAACT
                     GCGTGGCACCAATTTTCCGCCTGATGGTCCGGTTATGCAGAAAAAAACCATGGGTTGG
                     GAAGCAAGCACCGAACGTCTGTATCCTGAAGATGGCGTTCTGAAAGGTGATATCAAAA
                     TGGCACTGCGTCTGAAAGATGGCGGTCGTTATCTGGCAGATTTCAAAACCACCTACAA
                     AGCCAAAAAACCGGTTCAGATGCCTGGTGCATATAATGTTGATCGCAAACTGGATATT
                     ACCAGCCACAATGAAGATTATACCGTGGTGGAACAGTATGAACGTAGCGAAGGCCGTC
                     ATAGTACCGGTGGTATGGATGAACTGTACAAGGGCGGGAGCGGAGGCACGGGAGGTAC
                     C"
                     /note="Geneious type: Editing History Deletion"
     ORF             1360..>1794
                     /created_by="kaczmarc"
                     /label="cpGFP_original Nterm"
     misc_feature    1795..1809
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="2H12_C-linker"
     ORF             1810..2073
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD CTD 2"
     misc_feature    1948..1953
                     /created_by="kaczmarc"
                     /label="mutate"
     misc_feature    2079^2080
                     /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          <2086..>3890
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2111..2215
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             2216..3076
                     /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(2434..2453)
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      3247..3835
                     /direction="RIGHT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     3736..3755
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     trimmed         3889..3890
                     /Method="Max ambiguities: 2, Min 5' trim: 2, Min 3' trim:
                     2"
                     /label="Trimmed"
     misc_feature    3890^3891
                     /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 ggagggagca tggtgagcaa gggcgaggag
     1381 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1441 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1501 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1561 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1621 gccatgcccg aaggctacat tcaggagcgc accatcttct tcaaggacga cggcaactat
     1681 aagacacgcg ctgaggttaa gttcgagggc gacactctgg ttaaccgcat cgagctgaag
     1741 ggcatcgact tcaaggagga cggcaacatc ctgggccata agcttgaata taacatgttg
     1801 tcggagccgg agccgccgcc gaagctcgtc cttgacctgg agcgcctcgc gcacgtcccc
     1861 caggagaagg cgggcccgct ccagcgctac gcggcgacga tccagtcgca gcgcggcgac
     1921 tacaacggca aggtgctgtc gatccgtcag gacgacctgc gcaccctggc cgtgatctac
     1981 gaccagtcgc cctcggtcct caccgagcag ctgatcagct ggggcgtcct cgacgcggac
     2041 gcgcgccgcg ccgtcgccca cgaggagaac tgaactagtg acgtcaggtg gcacttttcg
     2101 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2161 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2221 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2281 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2341 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2401 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     2461 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2521 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2581 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2641 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2701 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     2761 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2821 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2881 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gctctcgcgg
     2941 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3001 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3061 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3121 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3181 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3241 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3301 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3361 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3421 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3481 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3541 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3601 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3661 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3721 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3781 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3841 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
//
