LOCUS       sequence-408082-        3071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PTac-YFP plasmid for LPattB2 integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3071)
  AUTHORS   Triassi AJ, Fields BD, Monahan CE, Means JM, Park Y, Doosthosseini 
            H, Padmakumar JP, Isabella VM, Voigt CA
  TITLE     Redesign of an Escherichia coli Nissle treatment for phenylketonuria
            using insulated genomic landing pads and genetic circuits to reduce 
            burden.
  JOURNAL   Cell Syst. 2023 Jun 21;14(6):512-524.e12. doi: 
            10.1016/j.cels.2023.05.004.
  PUBMED    37348465
REFERENCE   2  (bases 1 to 3071)
  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"; doi: 
            "10.1016/j.cels.2023.05"; journalName: "Cell Syst"; date: 
            "2023-06-21- 21"; volume: "14"; issue: "6"; pages: "512-524.e12"
COMMENT     Sequence Label: pAJT237-Ptac
FEATURES             Location/Qualifiers
     source          1..3071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(28..741,742..744,745..747))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(73..94)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     422..441
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     681..702
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             754..765
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(797..846)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    849..868
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(join(874..880,881..896,897..902))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(1000..1047)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1099..1190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1191..2006
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1191..2006
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1264..1283)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2017..2111
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     oriT            2262..2370
                     /note="incP origin of transfer"
     rep_origin      2414..2802
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gaatctcgcg tagacacata catgagctta cttgtacagc tcgtccatgc cgagagtgat
       61 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
      121 gctcagggcg gactggtagc taaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
      181 gattggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
      241 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
      301 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
      361 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
      421 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
      481 catggcggac ttgaagaagt cgtgcagctt catgtggtcg gggtagcggg cgaagcattg
      541 caggccgtag ccgaaggtgg taacgagggt gggccagggc acgggcagct tgcctgtggt
      601 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
      661 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
      721 gaacagctcc tcgcccttgc tcaccatcta gtatttctcc tctttctcta gtattaaaca
      781 aaattatttg tagaggctgt ttcgtcctca cggactcatc agaccggaaa gcacatccgg
      841 tgacagctaa ttgtgagcgc tcacaattcc acacattata cgagccgatg attaattgtc
      901 aacacagcca acgatcgttc tcctggtctt ccagtacaat ctgctctgat gccctcgagt
      961 gcggccgcac tagtgtcgac ggagctcgaa ttcggatccg aagttcctat actttctaga
     1021 gaataggaac ttcggaatag gaacttctat ttttataggt taatgtcatg ataataatgg
     1081 tttcttagac catgcctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1141 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagccata
     1201 ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat
     1261 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     1321 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1381 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1441 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
     1501 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1561 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1621 accgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1681 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1741 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1801 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1861 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1921 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1981 atttgatgct cgatgagttt ttctaaagct cgcttggact cctgttgata gatccagtaa
     2041 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     2101 ttggtgagaa tccaagctta aggtatactt tccgctgcat aaccctgctt cggggtcatt
     2161 atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg
     2221 gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta
     2281 ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc
     2341 aacgggaatc ctgctctgcg aggctggccg ataagctaca agaagatccg gccacgatgc
     2401 gtccggcgta gaggatctga agatcagcag ttcaacctgt tgatagtacg tactaagctc
     2461 tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt ttaacgaact
     2521 aaaccctcat ggctaacgta ctaagctctc atggctaacg tactaagctc tcatgtttca
     2581 cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct
     2641 ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg
     2701 tttaacggtt gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct
     2761 caaagcaatt ttgagtgaca caggaacact taacggctga catgggaatt agccatgggc
     2821 ccgtgcgaat catttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2881 cctgcgttat ccccggattc tgtggataac cgtattaccg cctggaccaa aacgaaaaaa
     2941 caccctttcg ggtgtctttt ctggaatttg gtaccgagcc tttggtcgaa aaaaaaagcc
     3001 cgcactgtca ggtgcgggct tttttctgtg tttccagcca agagcagtgc cttgcagaag
     3061 ttctcgggcg a
//
