LOCUS       sequence-408083-        3093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PBad-YFP plasmid for LPattB2 integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3093)
  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 3093)
  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: pAJT238-Pbad
FEATURES             Location/Qualifiers
     source          1..3093
                     /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    complement(1022..1069)
                     /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        1121..1212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1213..2028
                     /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             1213..2028
                     /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(1286..1305)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2039..2133
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     oriT            2284..2392
                     /note="incP origin of transfer"
     rep_origin      2436..2824
                     /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 tcacgagggt 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 tgacagctcg ggtatggaga aaaatataga gagttgcgat aaaaagcgtc aggtaggatc
      901 cgctaatctt atggataaaa atgctacagc caacgatcgt tctcctggtc ttccagtaca
      961 atctgctctg atgccctcga gtgcggccgc actagtgtcg acggagctcg aattcggatc
     1021 cgaagttcct atactttcta gagaatagga acttcggaat aggaacttct atttttatag
     1081 gttaatgtca tgataataat ggtttcttag accatgccta tttgtttatt tttctaaata
     1141 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1201 aaaaggaaga gtatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat
     1261 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1321 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1381 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1441 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1501 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     1561 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1621 tgtaattgtc cttttaacag cgaccgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1681 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1741 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     1801 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1861 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1921 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     1981 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaag ctcgcttgga
     2041 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     2101 ggttgccgcc gggcgttttt tattggtgag aatccaagct taaggtatac tttccgctgc
     2161 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     2221 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     2281 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     2341 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgataagcta
     2401 caagaagatc cggccacgat gcgtccggcg tagaggatct gaagatcagc agttcaacct
     2461 gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact
     2521 aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa
     2581 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat
     2641 aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat
     2701 aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca
     2761 ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct
     2821 gacatgggaa ttagccatgg gcccgtgcga atcattttta cggttcctgg ccttttgctg
     2881 gccttttgct cacatgttct ttcctgcgtt atccccggat tctgtggata accgtattac
     2941 cgcctggacc aaaacgaaaa aacacccttt cgggtgtctt ttctggaatt tggtaccgag
     3001 cctttggtcg aaaaaaaaag cccgcactgt caggtgcggg cttttttctg tgtttccagc
     3061 caagagcagt gccttgcaga agttctcggg cga
//
