LOCUS       sequence-386874-        8703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 PhiC31 cassette with constitutive promoter : 
            pPP2AA3:PhiC31-NLS:t35S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8703)
  AUTHORS   Guiziou S, Maranas CJ, Chu JC, Nemhauser JL
  TITLE     An integrase toolbox to record gene-expression during plant 
            development.
  JOURNAL   Nat Commun. 2023 Apr 3;14(1):1844. doi: 10.1038/s41467-023-37607-5.
  PUBMED    37012288
REFERENCE   2  (bases 1 to 8703)
  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: "Nat 
            Commun."; date: "2023-04-3"; volume: "14(1):1844. doi"; pages: " 
            10.1038/s41467-023-37607-5"
COMMENT     Sequence Label: L1P47
FEATURES             Location/Qualifiers
     source          1..8703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    257..279
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    287..463
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(695..1720)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(1740..2085)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1777..1797)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2288..2310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2302..2319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2303..2319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2326..2345
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2326..2344
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2927..2938
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2986..4800
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     CDS             4801..4821
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4859..5033
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5080..5104
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5195..5783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5275..5294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7292..8107)
                     /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             complement(7292..8107)
                     /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     8015..8034
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8099..8118
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(8398..8703,1..130)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 gtatgttcaa ttagcttttc aattatgaat atatatatca attattggtc gcccttggct
       61 tgtggacaat gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac
      121 cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt
      181 tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat
      241 ccccggaatt agatcttggc aggatatatt gtggtgtaac gttatcgatc tggattttag
      301 tactggattt tggttttagg aattagaaat tttattgata gaagtatttt acaaatacaa
      361 atacatacta agggtttctt atatgctcaa cacatgagcg aaaccctata agaaccctaa
      421 tttcccttat cgggaaacta ctcacacatt atttatggag aaaatagaga gagatagatt
      481 tgtagagaga gactggtgat ttcagcgtac cgaattaatt cctgaatcac tgcgaccggc
      541 cctcccgcga cccagccgag cgagcttagc gaactgtgga cgagaactgt gccaccaagc
      601 gtaaggccgt tctctcgcat ttgccttgct aggctcgcgc gagttgctgg ctgaggcgtt
      661 ctcgaaatca gctcttgttc ggtcggcatc tactctattc ctttgccctc ggacgagtgc
      721 tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg
      781 cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt
      841 cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga
      901 gttggtcaag accaatgcgg agcatatacg cccggagcct tggcgatcct gcaagctccg
      961 gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga
     1021 agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga
     1081 ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga
     1141 ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga
     1201 cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa
     1261 tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc
     1321 cgaacccgct cgtctggcta agatcggccg cagcgatagc atccatagcc tccgcgaccg
     1381 gctgaagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac
     1441 ggcgggagat gcaataggtc aggctctcgc tgaactcccc aatgtcaagc acttccggaa
     1501 tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg
     1561 cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag
     1621 attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa
     1681 acttctcgac agacgtcgcg gtgagttcag gctttttcat aggggggatc agcttgggct
     1741 gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg
     1801 gattgtgcgt catcccttac gtcagtggag atgtcacatc aatccacttg ctttgaagac
     1861 gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
     1921 ccactgtcgg cagaggcatc ttgaatgata gcctttcctt tatcgcaatg atggcatttg
     1981 taggagccac cttccttttc tactgtcctt tcgatgaagt gacagatagc tgggcaatgg
     2041 aatccgagga ggtttcccga aattaccctt tgttgaaaag tctcaatagc cctttggtct
     2101 tctgagactg tatctttgac atttttggag taggggtacg ataacattaa cgcttacaat
     2161 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     2221 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     2281 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     2341 tagggcttgg aggaggaaga agaagagagt tactgagaag cttcctttat acgagagaga
     2401 tattaaacga agtcgtattt gttggatata tgggccttta tgtattatac ataagcccat
     2461 ttagttctta tctttgacag gcccaattat atggacacag cttgaagatg gtttagctgc
     2521 tgcgaaagac gagctccggt ctgatttctc gttcttctga ttgatagatc gctcggaact
     2581 tggaaagcag cgtaatcggt aaattctcga tcatttcttc aatttcgaaa cattttccaa
     2641 ttgtatattg agaatctcaa atcgaattat tgtttagatc ttctctcttt attagtctaa
     2701 gaatggtaat agtttctctc tagattggga cggcacaaag tttcagttct ccattctaat
     2761 cattttttag aggctaatgg gtcacctttg aattatctca gaggcggaat tgagtttaaa
     2821 gtttgtaact ttatcgtctt ttgtattgat ggtgtctatg gcaaaggtga ttaactgcaa
     2881 aggtatgcat tgtgttttat tctgactttg tgtacatttg tgtttgattg aaggtaggga
     2941 gtggcaaagg tgattaactg caaaggattt gagttttggt gaggaatgga cacgtatgcc
     3001 ggcgcttatg atagacaatc aagagaaaga gagaactctt ctgccgcctc tccagctaca
     3061 cagagatctg ccaacgagga taaagctgct gatttacaaa gagaggtaga gagagatggg
     3121 ggtaggttta gatttgtggg tcatttctct gaggctcctg gtacatcagc ctttggtact
     3181 gctgaacggc ctgaatttga aagaatatta aatgagtgtc gggccggtcg actaaatatg
     3241 ataatagttt atgatgtatc taggttcagt agactcaaag ttatggatgc aattccgatc
     3301 gtgagcgagc tacttgcact cggtgttact atcgtaagta cacaagaagg tgtattcagg
     3361 cagggtaatg tgatggacct tatacacctt attatgcgac tcgacgctag ccataaagag
     3421 tcatcactga agagtgctaa aatcttggac accaagaacc tgcaaaggga gcttggaggc
     3481 tatgtaggag gaaaagctcc gtatggtttc gaactcgtga gtgagacaaa agagattacc
     3541 agaaatggta ggatggtgaa cgtggttatt aataagctag ctcatagcac aacccctctc
     3601 acagggcctt ttgagttcga acctgacgtt attagatggt ggtggagaga gatcaaaact
     3661 cacaaacatc ttccatttaa gccaggtagc caggctgcta tccaccctgg atctatcact
     3721 gggctttgta aaaggatgga tgctgacgca gtgccaactc gaggggaaac aatcggtaag
     3781 aagactgcat cttccgcttg ggatcccgct acagttatga gaatccttcg ggatccgcgt
     3841 attgctggat tcgccgcaga agtcatttat aagaaaaaac ctgatggtac tcctactaca
     3901 aaaatcgaag gttataggat ccaaagggat cctattactt tgcgtccagt tgaattagac
     3961 tgtggaccga tcatcgaacc tgccgagtgg tacgagttgc aggcctggct tgatggtaga
     4021 gggcgtggaa agggtttatc tcggggacag gcaattttgt cagcaatgga taagctttat
     4081 tgtgagtgtg gggctgttat gacatcaaaa cgtggagaag agtcaatcaa ggattcttac
     4141 aggtgcagac gtcgtaaagt ggtggaccct tctgctccag gacagcatga gggtacatgc
     4201 aacgtttcta tggctgctct tgataaattt gtggccgaac gtattttcaa caagattaga
     4261 cacgctgaag gggatgagga aactttagct cttctttggg aggctgcccg gcgttttggt
     4321 aagctaacag aggcaccaga aaaaagtggc gaaagagcta atttggtagc cgaaagagca
     4381 gacgccttga atgcattgga agaactttac gaagataggg ctgccggagc atatgatgga
     4441 ccagttggtc ggaagcattt caggaagcaa caagctgcac taactttgag acagcaaggc
     4501 gcagaggagc gactcgcaga gcttgaggcc gccgaggccc caaagcttcc actcgatcaa
     4561 tggtttcctg aggatgccga cgctgatcct acaggtccaa agagctggtg gggaagagct
     4621 agcgtagacg ataagagagt gtttgtgggt ctatttgttg acaagattgt agttactaag
     4681 tccactacgg gtcgaggaca gggcactccc atcgaaaaga gagccagtat tacctgggca
     4741 aagcctccaa ccgatgatga tgaagacgat gcacaagatg gtaccgagga tgtggcagca
     4801 ccaaaaaaaa agaggaaggt gtaagcttct ctagctagag tcgatcgaca agctcgagtt
     4861 tctccataat aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc
     4921 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
     4981 atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag atcgctacca
     5041 cagaatcagg ggataacgca ggaaagaaca tgaaggcctt gacaggatat attggcgggt
     5101 aaactaagtc gctgtatgtg tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg
     5161 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5221 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5281 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5341 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5401 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5461 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5521 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5581 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5641 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aagaagagtt ggtagctctt
     5701 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtact gagagatccc
     5941 ctcataattt ccccaaagcg taaccatgtg tgaataaatt ttgagctagt agggttgcag
     6001 ccacgagtaa gtcttccctt gttattgtgt agccagaatg ccgcaaaact tccatgccta
     6061 agcgaactgt tgagagtacg tttcgatttc tgactgtgtt agcctggaag tgcttgtccc
     6121 aaccttgttt ctgagcatga acgcccgcaa gccaacatgt tagttgaagc atcagggcga
     6181 ttagcagcat gatatcaaaa cgctctgagc tgctcgttcg gctatggcgt aggcctagtc
     6241 cgtaggcagg acttttcaag tctcggaagg tttcttcaat ctgcattcgc ttcgaataga
     6301 tattaacaag ttgtttgggt gttcgaattt caacaggtaa gttagttgct agaacccatg
     6361 gctcctttgc cgacgctgag tagattttag gtgacgggtg gtgacaatga gtccgtgtcg
     6421 agcgctgatt ttttcggcct ttagagcgag atttatacaa tagaatttgg catgagattg
     6481 gattgctttt agtcagcctc ttatagccta aagtctttga gtgactagat gacatatcat
     6541 gtaagttgct gataggtttc cagttttccg ctcctaggtc tgcatattgt acttttcctc
     6601 ttactcgact taaccagtac caacccagct tctcaacgga tttataccat ggcactttaa
     6661 agccagcatc actgacaatg agcggtgtgg tgttactcgg tagaatgctc gcaaggtcgg
     6721 ctagaaattg gtcatgagct ttctttgaac attgctctga aagcgggaac gctttctcat
     6781 aaagagtaac agaacgaccg tgtagtgcga ctgaagctcg caataccata agtcgttttt
     6841 gctcacgaat atcagaccag tcaacaagta caatgggcat cgtattgccc gaacagataa
     6901 agctagcatg ccaacggtat acagcgagtc gctctttgtg gaggtgacga ttacctaaca
     6961 atcggtcgat tcgtttgatg ttatgttttg ttctcgcttt ggttggcagg ttacggccaa
     7021 gttcggtaag agtgagagtt ttacagtcaa gtaatgcgtg gcaagccaac gttaagctgt
     7081 tgagtcgttt taagtgtaat tcggggcaga attggtaaag agagtcgtgt aaaatatcga
     7141 gttcgcacat cttgttgtct gattattgat ttttcgcgaa accatttgat catatgacaa
     7201 gatgtgtatc caccttaact taatgatttt taccaaaatc attaggggat tcatcagtct
     7261 aaagtatata tgtgtaacat tggtctagtg attagaaaaa ctcatcgagc atcaaatgaa
     7321 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     7381 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     7441 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     7501 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     7561 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     7621 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     7681 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     7741 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     7801 ctgggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     7861 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     7921 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     7981 atcggtagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     8041 aatcagcatc catgttggaa tttaatcgcg gccttgagca agacgtttcc cgttgaatat
     8101 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     8161 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt
     8221 gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag
     8281 accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc
     8341 tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag gcgatcccca
     8401 tccaacagcc cgccgtcgag cgggcttttt tatccccgga agcctgtgga tagagggtag
     8461 ttatccacgt gaaaccgcta atgccccgca aagccttgat tcacggggct ttccggcccg
     8521 ctccaaaaac tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct aatcggagta
     8581 cgtgaaatcg ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta
     8641 atagcccttt cagatcaaca gcttgcaaac acccctcgct ccggcaagta gttacagcaa
     8701 gta
//
