LOCUS       sequence-229509-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-noise, tetracycline inducible expression of tdPP7cp-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  TITLE     Plasmids for mRNA detection with PP7 in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4968)
  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            "Unpublished"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_tdPP7cp_mCherry
FEATURES             Location/Qualifiers
     source          1..4968
                     /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      1045..1054
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1785..2408
                     /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(1858..1877)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2453..2499
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2518..2534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2518..2534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2531..2553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2542..2558
                     /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(2566..2572,2573..2590,2591..2596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2611..2632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3172..3717)
                     /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(3252..3271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3908..4699,4700..4768))
                     /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     4531..4550
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4769..4873)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4941..4959
                     /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 gggccgcgca
      661 gatccgctgg catctaaaac catcgttctg tccgtgggcg aagcgacccg taccctgact
      721 gagatccagt ctaccgcgga tcgccagatt tttgaagaaa aggtaggccc gctggttggc
      781 cgcctgcgtc tgaccgcgag cctgcgccag aacggtgcaa aaaccgctta tcgtgtgaac
      841 ctgaagctgg atcaggccga tgttgtagac agcggcctgc cgaaggtacg ttacactcag
      901 gtttggtccc acgatgttac catcgtggca aactctaccg aagctagtcg taaaagcctg
      961 tacgatctca cgaaatccct ggtcgctacc tcccaggtgg aagatctggt ggttaacctg
     1021 gttccactgg gtcgcccgcc ggttgccacc atggtgtcta aaggcgaaga agataatatg
     1081 gcgatcatca aagagttcat gcgctttaag gtgcacatgg agggctccgt aaacggtcac
     1141 gagttcgaaa ttgaagggga aggcgaaggc cgtccgtacg aaggcaccca gaccgcaaaa
     1201 ctgaaggtca ctaaaggtgg cccgctgccg ttcgcgtggg atatcctctc tccgcagttc
     1261 atgtacggtt ctaaagcgta cgttaaacac ccagctgaca tcccggacta cctgaaactc
     1321 tccttcccgg aaggctttaa gtgggaacgc gtcatgaatt tcgaggacgg cggtgtcgtc
     1381 accgttactc aggactcctc tctccaggac ggtgaattta tctacaaagt aaagctgcgc
     1441 ggtaccaact ttccgagcga tggtccggtg atgcaaaaga aaaccatggg ttgggaagcg
     1501 tcctctgaac gtatgtaccc agaagatggt gcactcaaag gcgaaattaa acagcgtctg
     1561 aagctgaaag atggtggtca ttacgacgcc gaagttaaaa ccacctacaa ggcaaagaag
     1621 ccggttcagc tcccaggtgc ctacaacgtg aacattaagc tggacatcac cagccataac
     1681 gaagactaca ccatcgtcga acagtacgaa cgtgccgagg gccgccactc cactggcggc
     1741 atggatgaac tgtataagta ataatctagc aggaggattt caccatgtct agattagata
     1801 aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa
     1861 caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa
     1921 aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc
     1981 acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt
     2041 ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta
     2101 cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt
     2161 cactagagaa tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat
     2221 tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta
     2281 tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct
     2341 tcttattcgg ccttgaattg atcatctgcg gattagaaaa acaacttaaa tgtgaaagtg
     2401 ggtcttaata actgcagccc gggggatccc atggtacgcg tgctagaggc atcaaataaa
     2461 acgaaaggct cagtcgaaag actgggcctt tcgttttata agcttggcgt aatcatggtc
     2521 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2581 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2641 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2701 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2761 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2881 aaaggccagg aaccgtaaaa aggcgcgcta gcggagtgta tactggctta ctatgttggc
     2941 actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc
     3001 gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct
     3061 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
     3121 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     3181 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     3241 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     3301 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     3361 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     3421 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     3481 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     3541 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     3601 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     3661 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     3721 gatcatctta ttaatcagat aaaatataga tcttttctac ggggtctgac gctcagtgga
     3781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3901 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3961 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4021 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4081 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4141 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4201 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4261 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4321 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4381 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4441 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4501 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4561 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4621 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4681 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4741 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4801 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4861 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4921 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
