LOCUS       sequence-195303-        3576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in the presence of Pseudomonas fluorescence sigma 
            factor ECF20_992. Repressed by TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3576)
  TITLE     Unpublished Murray Lab Plasmids
REFERENCE   2  (bases 1 to 3576)
  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"
COMMENT     Sequence Label: P20_992-tetO-deGFP
FEATURES             Location/Qualifiers
     source          1..3576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     487..506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(725..829)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     897..915
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    1258..1276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1283..1305
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1342..1363)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1603..1622)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1950..1971
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2510..2527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2681..3269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2761..2780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3440..3576,1..655,656..724))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cgcgtgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaagtg
       61 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      121 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      181 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      241 gatcaaggcg agttgcatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      301 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      361 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      421 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
      481 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
      541 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
      601 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
      661 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      841 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
      901 ggcgtatcac gaggcccttt cgtgttcaag aattctggcg aatcctctga ccagccagaa
      961 aacgaccttt ctgtggtgaa accggatgct gcaattcaga gcgccagcaa gtgggggaca
     1021 gcagatgacc tgaccgccgc agagtggatg tttgacatgg tgatgactat cgcaccatca
     1081 gccagaaaac cgaattttgc tgggtgggct aacgatatcc gcctgatgcg tgaacgtgac
     1141 ggacgtaacg aagacaacca cgcattgctg ttcgcgcgga taaaaatttc atttgcccgc
     1201 gacggattcc ccgcccatct atcgttgaac ccatcagctg cgttcatcag cgaaagctcc
     1261 ctatcagtga tagagaaata attttgttta actttaagaa ggagatatac aatggagctt
     1321 ttcactggcg ttgttcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1381 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1441 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1501 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1561 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1621 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1681 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1741 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1801 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1861 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1921 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1981 gggatctaat gaactaacgc atgagaaagc cccggaagat caccttccgg gggctttttt
     2041 attgcgccct tgagagtcgg gcattgtctt cgctccttcc ggtgggcgcg gggcaaacca
     2101 ggtggttggc caggttgagt tggtgcgttt gttgaggatg tcatgcacat ccttcggcaa
     2161 ttccacgctc cagccttccg cgatttgcag taccggtccc aggcctttga tgatgttgac
     2221 acgagtcatg gtgaacgtga cggacgtaac gaagacaacc acgcattgct gttcgcgcgg
     2281 ataaaaattt catttgcccg cgacggattc cccgccggga tctaatgaac taacgcatga
     2341 gacccccgga agatcacctt ccgggggctt ttttattgcg cccttgagag tcgggcattg
     2401 tcttcgctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtgtt
     2461 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     2521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3541 agggcttacc atctggcccc agtgctgcaa tgatac
//
