LOCUS       sequence-229501-        4977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-noise, tetracycline inducible expression of tdPP7cp-mCerulean3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4977)
  TITLE     Plasmids for mRNA detection with PP7 in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4977)
  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            "Unpublished"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_mCerulean3
FEATURES             Location/Qualifiers
     source          1..4977
                     /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             join(1051..1053,1054..1056,1057..1770)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1096..1117)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1357..1376)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1704..1725
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1794..2417
                     /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(1867..1886)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2462..2508
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2527..2543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2527..2543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2540..2562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2551..2567
                     /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(2575..2581,2582..2599,2600..2605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2620..2641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3181..3726)
                     /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(3261..3280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3917..4708,4709..4777))
                     /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     4540..4559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4778..4882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4950..4968
                     /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 cattgttctg 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 atggtgagca agggcgagga gctgttcacc
     1081 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1141 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1201 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagctg gggcgtgcag
     1261 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1321 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1381 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1441 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat ccacggcaac
     1501 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcgg cctcaactgc
     1561 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1621 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     1681 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1741 actctcggca tggacgagct gtacaagtaa taatctagca ggaggatttc accatgtcta
     1801 gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     1861 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     1921 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     1981 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
     2041 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     2101 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     2161 aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag
     2221 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     2281 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     2341 agccagcctt cttattcggc cttgaattga tcatctgcgg attagaaaaa caacttaaat
     2401 gtgaaagtgg gtcttaataa ctgcagcccg ggggatccca tggtacgcgt gctagaggca
     2461 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttataa gcttggcgta
     2521 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2581 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2641 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2701 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2761 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2821 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2881 aggccagcaa aaggccagga accgtaaaaa ggcgcgctag cggagtgtat actggcttac
     2941 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     3001 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     3061 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     3121 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     3181 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     3241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     3301 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     3361 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     3421 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3481 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     3541 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     3601 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     3661 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     3721 tctcaagaag atcatcttat taatcagata aaatatagat cttttctacg gggtctgacg
     3781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4921 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
