LOCUS       sequence-229514-        4599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-noise, tetracycline inducible expression of mPP7cp-mCerulean3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4599)
  TITLE     Plasmids for mRNA detection with PP7 in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4599)
  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            "Unpublished"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_mCerulean3
FEATURES             Location/Qualifiers
     source          1..4599
                     /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             join(673..675,676..678,679..1392)
                     /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(718..739)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(979..998)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1326..1347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1416..2039
                     /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(1489..1508)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2084..2130
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2149..2165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2149..2165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2162..2184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2173..2189
                     /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(2197..2203,2204..2221,2222..2227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2242..2263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2803..3348)
                     /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(2883..2902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3539..4330,4331..4399))
                     /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     4162..4181
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4400..4504)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4572..4590
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      721 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      781 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      841 ccctggccca ccctcgtgac caccctgagc tggggcgtgc agtgcttcgc ccgctacccc
      901 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      961 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1021 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1081 atcctggggc acaagctgga gtacaacgcc atccacggca acgtctatat caccgccgac
     1141 aagcagaaga acggcatcaa ggccaacttc ggcctcaact gcaacatcga ggacggcagc
     1201 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1261 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1321 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1381 ctgtacaagt aataatctag caggaggatt tcaccatgtc tagattagat aaaagtaaag
     1441 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
     1501 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
     1561 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
     1621 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
     1681 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
     1741 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
     1801 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc
     1861 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
     1921 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
     1981 gccttgaatt gatcatctgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaat
     2041 aactgcagcc cgggggatcc catggtacgc gtgctagagg catcaaataa aacgaaaggc
     2101 tcagtcgaaa gactgggcct ttcgttttat aagcttggcg taatcatggt catagctgtt
     2161 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2221 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2281 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2521 gaaccgtaaa aaggcgcgct agcggagtgt atactggctt actatgttgg cactgatgag
     2581 ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga
     2641 atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc
     2701 gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga
     2761 agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc
     2821 ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact
     2881 ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt
     2941 tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac
     3001 tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt
     3061 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa
     3121 aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc
     3181 gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta
     3241 cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt
     3301 ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt
     3361 attaatcaga taaaatatag atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3421 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3481 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3541 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3601 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3661 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3721 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3781 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3841 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3901 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3961 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4021 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4081 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4141 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4201 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4261 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4321 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4381 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4441 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4501 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4561 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
