LOCUS       sequence-386302-        8999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 PhiC31 cassette with constitutive promoter: 
            pPP2AA3-PhiC31-NLS-tUBQ1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8999)
  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 8999)
  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: L1P2
FEATURES             Location/Qualifiers
     source          1..8999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2060..2875)
                     /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(2060..2875)
                     /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     2783..2802
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2867..2886
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3166..3601
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    3728..3750
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    3758..3934
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4166..5191)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(5211..5556)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(5248..5268)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     5759..5781
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5773..5790
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5774..5790
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5797..5816
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5797..5815
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6398..6409
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6457..8271
                     /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             8272..8292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    8847..8871
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(join(8962..8999,1..551))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
       61 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      421 gcgctctgct gaagccagtt accttcggaa gaagagttgg tagctcttga tccggcaaac
      481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      661 taaattaaaa atgaagtttt aaatcaatct aaagtactga tgaatcccct aatgattttg
      721 gtaaaaatca ttaagttaag gtggatacac atcttgtcat atgatcaaat ggtttcgcga
      781 aaaatcaata atcagacaac aagatgtgcg aactcgatat tttacacgac tctctttacc
      841 aattctgccc cgaattacac ttaaaacgac tcaacagctt aacgttggct tgccacgcat
      901 tacttgactg taaaactctc actcttaccg aacttggccg taacctgcca accaaagcga
      961 gaacaaaaca taacatcaaa cgaatcgacc gattgttagg taatcgtcac ctccacaaag
     1021 agcgactcgc tgtataccgt tggcatgcta gctttatctg ttcgggcaat acgatgccca
     1081 ttgtacttgt tgactggtct gatattcgtg agcaaaaacg acttatggta ttgcgagctt
     1141 cagtcgcact acacggtcgt tctgttactc tttatgagaa agcgttcccg ctttcagagc
     1201 aatgttcaaa gaaagctcat gaccaatttc tagccgacct tgcgagcatt ctaccgagta
     1261 acaccacacc gctcattgtc agtgatgctg gctttaaagt gccatggtat aaatccgttg
     1321 agaagctggg ttggtactgg ttaagtcgag taagaggaaa agtacaatat gcagacctag
     1381 gagcggaaaa ctggaaacct atcagcaact tacatgatat gtcatctagt cactcaaaga
     1441 ctttaggcta taagaggctg actaaaagca atccaatctc atgccaaatt ctattgtata
     1501 aatctcgctc taaaggccga aaaaatcagc gctcgacacg gactcattgt caccacccgt
     1561 cacctaaaat ctactcagcg tcggcaaagg agccatgggt tctagcaact aacttacctg
     1621 ttgaaattcg aacacccaaa caacttgtta atatctattc gaagcgaatg cagattgaag
     1681 aaaccttccg agacttgaaa agtcctgcct acggactagg cctacgccat agccgaacga
     1741 gcagctcaga gcgttttgat atcatgctgc taatcgccct gatgcttcaa ctaacatgtt
     1801 ggcttgcggg cgttcatgct cagaaacaag gttgggacaa gcacttccag gctaacacag
     1861 tcagaaatcg aaacgtactc tcaacagttc gcttaggcat ggaagttttg cggcattctg
     1921 gctacacaat aacaagggaa gacttactcg tggctgcaac cctactagct caaaatttat
     1981 tcacacatgg ttacgctttg gggaaattat gaggggatct ctcagtctaa agtatatatg
     2041 tgtaacattg gtctagtgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     2101 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     2161 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     2221 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     2281 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     2341 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     2401 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     2461 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     2521 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgcag
     2581 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     2641 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     2701 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cggtagattg
     2761 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     2821 tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg ctcataacac
     2881 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     2941 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     3001 cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg
     3061 caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg
     3121 gctccctcac tttctggctg gatgatgggg cgattcaggc gatccccatc caacagcccg
     3181 ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt atccacgtga
     3241 aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct ccaaaaacta
     3301 tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg tgaaatcgct
     3361 aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat agccctttca
     3421 gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt agtatgttca
     3481 attagctttt caattatgaa tatatatatc aattattggt cgcccttggc ttgtggacaa
     3541 tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca ccgtcgagcg
     3601 cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt ttttttttga
     3661 aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatttccga tccccggaat
     3721 tagatcttgg caggatatat tgtggtgtaa cgttatcgat ctggatttta gtactggatt
     3781 ttggttttag gaattagaaa ttttattgat agaagtattt tacaaataca aatacatact
     3841 aagggtttct tatatgctca acacatgagc gaaaccctat aagaacccta atttccctta
     3901 tcgggaaact actcacacat tatttatgga gaaaatagag agagatagat ttgtagagag
     3961 agactggtga tttcagcgta ccgaattaat tcctgaatca ctgcgaccgg ccctcccgcg
     4021 acccagccga gcgagcttag cgaactgtgg acgagaactg tgccaccaag cgtaaggccg
     4081 ttctctcgca tttgccttgc taggctcgcg cgagttgctg gctgaggcgt tctcgaaatc
     4141 agctcttgtt cggtcggcat ctactctatt cctttgccct cggacgagtg ctggggcgtc
     4201 ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc
     4261 gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac
     4321 cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa
     4381 gaccaatgcg gagcatatac gcccggagcc ttggcgatcc tgcaagctcc ggatgcctcc
     4441 gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt
     4501 tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta
     4561 tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga
     4621 cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac
     4681 tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata
     4741 tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc
     4801 tcgtctggct aagatcggcc gcagcgatag catccatagc ctccgcgacc ggctgaagaa
     4861 cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga
     4921 tgcaataggt caggctctcg ctgaactccc caatgtcaag cacttccgga atcgggagcg
     4981 cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat
     5041 ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga gattcttcgc
     5101 cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga aacttctcga
     5161 cagacgtcgc ggtgagttca ggctttttca taggggggat cagcttgggc tgtcctctcc
     5221 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg
     5281 tcatccctta cgtcagtgga gatgtcacat caatccactt gctttgaaga cgtggttgga
     5341 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
     5401 gcagaggcat cttgaatgat agcctttcct ttatcgcaat gatggcattt gtaggagcca
     5461 ccttcctttt ctactgtcct ttcgatgaag tgacagatag ctgggcaatg gaatccgagg
     5521 aggtttcccg aaattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
     5581 gtatctttga catttttgga gtaggggtac gataacatta acgcttacaa tttccattcg
     5641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5701 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5761 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcttg
     5821 gaggaggaag aagaagagag ttactgagaa gcttccttta tacgagagag atattaaacg
     5881 aagtcgtatt tgttggatat atgggccttt atgtattata cataagccca tttagttctt
     5941 atctttgaca ggcccaatta tatggacaca gcttgaagat ggtttagctg ctgcgaaaga
     6001 cgagctccgg tctgatttct cgttcttctg attgatagat cgctcggaac ttggaaagca
     6061 gcgtaatcgg taaattctcg atcatttctt caatttcgaa acattttcca attgtatatt
     6121 gagaatctca aatcgaatta ttgtttagat cttctctctt tattagtcta agaatggtaa
     6181 tagtttctct ctagattggg acggcacaaa gtttcagttc tccattctaa tcatttttta
     6241 gaggctaatg ggtcaccttt gaattatctc agaggcggaa ttgagtttaa agtttgtaac
     6301 tttatcgtct tttgtattga tggtgtctat ggcaaaggtg attaactgca aaggtatgca
     6361 ttgtgtttta ttctgacttt gtgtacattt gtgtttgatt gaaggtaggg agtggcaaag
     6421 gtgattaact gcaaaggatt tgagttttgg tgaggaatgg acacgtatgc cggcgcttat
     6481 gatagacaat caagagaaag agagaactct tctgccgcct ctccagctac acagagatct
     6541 gccaacgagg ataaagctgc tgatttacaa agagaggtag agagagatgg gggtaggttt
     6601 agatttgtgg gtcatttctc tgaggctcct ggtacatcag cctttggtac tgctgaacgg
     6661 cctgaatttg aaagaatatt aaatgagtgt cgggccggtc gactaaatat gataatagtt
     6721 tatgatgtat ctaggttcag tagactcaaa gttatggatg caattccgat cgtgagcgag
     6781 ctacttgcac tcggtgttac tatcgtaagt acacaagaag gtgtattcag gcagggtaat
     6841 gtgatggacc ttatacacct tattatgcga ctcgacgcta gccataaaga gtcatcactg
     6901 aagagtgcta aaatcttgga caccaagaac ctgcaaaggg agcttggagg ctatgtagga
     6961 ggaaaagctc cgtatggttt cgaactcgtg agtgagacaa aagagattac cagaaatggt
     7021 aggatggtga acgtggttat taataagcta gctcatagca caacccctct cacagggcct
     7081 tttgagttcg aacctgacgt tattagatgg tggtggagag agatcaaaac tcacaaacat
     7141 cttccattta agccaggtag ccaggctgct atccaccctg gatctatcac tgggctttgt
     7201 aaaaggatgg atgctgacgc agtgccaact cgaggggaaa caatcggtaa gaagactgca
     7261 tcttccgctt gggatcccgc tacagttatg agaatccttc gggatccgcg tattgctgga
     7321 ttcgccgcag aagtcattta taagaaaaaa cctgatggta ctcctactac aaaaatcgaa
     7381 ggttatagga tccaaaggga tcctattact ttgcgtccag ttgaattaga ctgtggaccg
     7441 atcatcgaac ctgccgagtg gtacgagttg caggcctggc ttgatggtag agggcgtgga
     7501 aagggtttat ctcggggaca ggcaattttg tcagcaatgg ataagcttta ttgtgagtgt
     7561 ggggctgtta tgacatcaaa acgtggagaa gagtcaatca aggattctta caggtgcaga
     7621 cgtcgtaaag tggtggaccc ttctgctcca ggacagcatg agggtacatg caacgtttct
     7681 atggctgctc ttgataaatt tgtggccgaa cgtattttca acaagattag acacgctgaa
     7741 ggggatgagg aaactttagc tcttctttgg gaggctgccc ggcgttttgg taagctaaca
     7801 gaggcaccag aaaaaagtgg cgaaagagct aatttggtag ccgaaagagc agacgccttg
     7861 aatgcattgg aagaacttta cgaagatagg gctgccggag catatgatgg accagttggt
     7921 cggaagcatt tcaggaagca acaagctgca ctaactttga gacagcaagg cgcagaggag
     7981 cgactcgcag agcttgaggc cgccgaggcc ccaaagcttc cactcgatca atggtttcct
     8041 gaggatgccg acgctgatcc tacaggtcca aagagctggt ggggaagagc tagcgtagac
     8101 gataagagag tgtttgtggg tctatttgtt gacaagattg tagttactaa gtccactacg
     8161 ggtcgaggac agggcactcc catcgaaaag agagccagta ttacctgggc aaagcctcca
     8221 accgatgatg atgaagacga tgcacaagat ggtaccgagg atgtggcagc accaaaaaaa
     8281 aagaggaagg tgtaagctta gagactctta tcaagaatcc catctcttgc ttgctttttt
     8341 ttgttgtctt ccctttgata gggtttgttt ttcttgtttc agtgactttc tatgttaaaa
     8401 gataatgtca gtaaaaggat ttggttttct attattctga atcgattacg gaagattctt
     8461 gcttaattcc aatctataca agtatcgtga aataatgacc gtttatgtga ttaggagacg
     8521 tgtttcatta ataaaatata agatcaatac attgttagta gtgataaact atgtacaaat
     8581 tgtattgatt gtaaaagaaa cacaataggt tccttttttc tacaatatat tgtgacagac
     8641 tctctgtttt aacgaatgaa ttaaatttgt caacgaaaaa attaaagaat gatgtgatca
     8701 ctttgcacct tggttcatct tcagaggctt ctgcatgttc catgcattga ggctctctac
     8761 ggtcacaggt tgtgagccgt tgttaaagac aaacaaatgc gctaccacag aatcagggga
     8821 taacgcagga aagaacatga aggccttgac aggatatatt ggcgggtaaa ctaagtcgct
     8881 gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     8941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaa
//
