LOCUS       sequence-182397-        3162 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses TetR in the presence of Pseudomonas fluorescence sigma 
            factor ECF20_992.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3162)
  TITLE     Unpublished Murray Lab Plasmids
REFERENCE   2  (bases 1 to 3162)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: P20_992-TetR
FEATURES             Location/Qualifiers
     source          1..3162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             212..234
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             241..864
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAKKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(314..333)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1052..1069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1223..1811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1303..1322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1982..2773,2774..2842))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRCIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2605..2624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2843..2947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3015..3033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tggatgtttg acatggtgat gactatcgca ccatcagcca gaaaaccgaa ttttgctggg
       61 tgggctaacg atatccgcct gatgcgtgaa cgtgacggac gtaacgaaga caaccacgca
      121 tgcgcggata aaaatttcat ttgcccgcga cggattcccc gcccatctat cgttgaaccc
      181 atcagctgcg ttcatcagcg aaagcaataa ttttgtttaa ctttaagaag gagatataca
      241 atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
      301 ggaatcgaag gtttaacaac ccgtaaactc gccaagaagc taggtgtaga gcagcctaca
      361 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
      421 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
      481 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
      541 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
      601 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt
      661 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
      721 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
      781 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
      841 cttaaatgtg aaagtgggtc ttaatgaact aacgcatgag aaagcccccg gaagatcacc
      901 ttccgggggc ttttttattg cgcccttgag agtcgggcat tgtcttcgct ccttccggtg
      961 ggcgcggggc atgactatcg tcgccgcact tatgactgtg ttctttatca tgcaactcgt
     1021 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2101 ccagtgctgc aatgatacag cgtgacccac gctcaccggc tccagattta tcagcaataa
     2161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2341 tcagctccgg ttcccaacga tcaaggcgag ttgcatgatc ccccatgttg tgcaaaaaag
     2401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2581 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2941 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     3001 cattaaccta taaaaatagg cgtatcacga ggccctttcg tgttcaagaa ttctggcgaa
     3061 tcctctgacc agccagaaaa cgacctttct gtggtgaaac cggatgctgc aattcagagc
     3121 gccagcaagt gggggacagc agatgacctg accgccgcag ag
//
