LOCUS       sequence-229515-        4590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-noise, tetracycline inducible expression of mPP7cp-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4590)
  TITLE     Plasmids for mRNA detection with PP7 in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4590)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pZH_mPP7cp_mCherry
FEATURES             Location/Qualifiers
     source          1..4590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(189..207,208..213,214..230,231..236,237..262)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    214..232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             272..283
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1407..2030
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1480..1499)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2075..2121
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2140..2156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2140..2156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2153..2175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2164..2180
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2188..2194,2195..2212,2213..2218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2233..2254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2794..3339)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2874..2893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3530..4321,4322..4390))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4153..4172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4391..4495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4563..4581
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgtc cctatcagtg atagagattg acatccctat cagtgataga gatactgagc
      241 acatcagcag gacgcactga ccgaattcat taaagaggag aaaggtaccg catgtctaaa
      301 actatcgtgc tgtctgttgg tgaagcaacc cgcaccctga ctgaaattca gtctactgct
      361 gatcgccaaa tcttcgaaga aaaagttggt ccgctggttg gccgtctgcg tctgactgct
      421 tccctgcgtc agaacggcgc gaagaccgcg tatcgcgtta atctgaaact ggatcaagcg
      481 gacgtagtgg actctggtct gccgaaagtt cgctacacgc aggtgtggtc tcacgacgtt
      541 accatcgttg ctaattccac cgaagccagc cgtaagtccc tctacgacct caccaagagc
      601 ctggttgcaa cctctcaggt tgaggatctg gtggtcaacc tggtaccact gggccgcccg
      661 ccggttgcca ccatggtgtc taaaggcgaa gaagataata tggcgatcat caaagagttc
      721 atgcgcttta aggtgcacat ggagggctcc gtaaacggtc acgagttcga aattgaaggg
      781 gaaggcgaag gccgtccgta cgaaggcacc cagaccgcaa aactgaaggt cactaaaggt
      841 ggcccgctgc cgttcgcgtg ggatatcctc tctccgcagt tcatgtacgg ttctaaagcg
      901 tacgttaaac acccagctga catcccggac tacctgaaac tctccttccc ggaaggcttt
      961 aagtgggaac gcgtcatgaa tttcgaggac ggcggtgtcg tcaccgttac tcaggactcc
     1021 tctctccagg acggtgaatt tatctacaaa gtaaagctgc gcggtaccaa ctttccgagc
     1081 gatggtccgg tgatgcaaaa gaaaaccatg ggttgggaag cgtcctctga acgtatgtac
     1141 ccagaagatg gtgcactcaa aggcgaaatt aaacagcgtc tgaagctgaa agatggtggt
     1201 cattacgacg ccgaagttaa aaccacctac aaggcaaaga agccggttca gctcccaggt
     1261 gcctacaacg tgaacattaa gctggacatc accagccata acgaagacta caccatcgtc
     1321 gaacagtacg aacgtgccga gggccgccac tccactggcg gcatggatga actgtataag
     1381 taataatcta gcaggaggat ttcaccatgt ctagattaga taaaagtaaa gtgattaaca
     1441 gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc
     1501 agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag cgggctttgc
     1561 tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc cctttagaag
     1621 gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt gctttactaa
     1681 gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa cagtatgaaa
     1741 ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag aatgcattat
     1801 atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat caagagcatc
     1861 aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca ttattacgac
     1921 aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc ggccttgaat
     1981 tgatcatctg cggattagaa aaacaactta aatgtgaaag tgggtcttaa taactgcagc
     2041 ccgggggatc ccatggtacg cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa
     2101 agactgggcc tttcgtttta taagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2161 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2221 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2281 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2341 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2521 aaaggcgcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     2581 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     2641 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     2701 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     2761 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     2821 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     2881 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     2941 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     3001 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     3061 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     3121 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     3181 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     3241 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     3301 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     3361 ataaaatata gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3421 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3481 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3541 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3601 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3661 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3721 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3781 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3841 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3901 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3961 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4021 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4081 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4141 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4201 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4261 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4321 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4381 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4441 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4501 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4561 aaaataggcg tatcacgagg ccctttcgtc
//
