LOCUS       p2H12ref                4598 bp ds-DNA     circular SYN 14-MAY-2024
DEFINITION  E. coli plasmid with 2H12 sensor with a mScarlet-I operon fusion as
            reference FP. UJ11206.
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..>2793
                     /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"
     ORF             778..1047
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD-M1"
     CDS             778..2073
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="2H12 (cdGreen)"
     ORF             1069..>1797
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="cpGFP"
     misc_feature    1231
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     ORF             1810..2073
                     /created_by="kaczmarc"
                     /modified_by="kaczmarc"
                     /label="BldD-M2"
     CDS             2080..2781
                     /created_by="kaczmarc"
                     /label="mScarlet-I"
     misc_feature    2659
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     source          <2794..>4598
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2819..2923
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             2924..3784
                     /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(3142..3161)
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      3955..4543
                     /direction="RIGHT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     4444..4463
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     misc_feature    4598^4599
                     /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 tgaactagta tgagcaagaa atatggtgaa
     2101 gccgtgatca aagaatttat gcgctttaaa gttcacatgg aaggtagcat gaatggccac
     2161 gaatttgaaa ttgaaggtga aggcgaaggt cgtccgtatg aaggcaccca gaccgcaaaa
     2221 ctgaaagtta ccaaaggtgg tccgctgccg tttagctggg atattctgag tccgcagttt
     2281 atgtatggta gccgtgcatt tatcaaacat ccggcagata ttccggatta ctataaacag
     2341 agctttccgg aaggttttaa atgggaacgt gtgatgaatt ttgaagatgg tggtgcagtt
     2401 accgttacac aggataccag cctggaagat ggcaccctga tctataaagt taaactgcgt
     2461 ggcaccaatt ttccgcctga tggtccggtt atgcagaaaa aaaccatggg ttgggaagca
     2521 agcaccgaac gtctgtatcc tgaagatggc gttctgaaag gtgatatcaa aatggcactg
     2581 cgtctgaaag atggcggtcg ttatctggca gatttcaaaa ccacctacaa agccaaaaaa
     2641 ccggttcaga tgcctggtgc atataatgtt gatcgcaaac tggatattac cagccacaat
     2701 gaagattata ccgtggtgga acagtatgaa cgtagcgaag gccgtcatag taccggtggt
     2761 atggatgaac tgtacaagtg agctagcgac gtcaggtggc acttttcggg gaaatgtgcg
     2821 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2881 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2941 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3001 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3061 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3121 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3181 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3241 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3301 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3361 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3421 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     3481 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3541 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3601 ctggtttatt gctgataaat ctggagccgg tgagcgtggc tctcgcggta tcattgcagc
     3661 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3721 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3781 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4081 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4561 ctttttacgg ttcctggcct tttgctggcc ttttgctc
//
