LOCUS       sequence-386333-        7643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 PhiC31 cassette with constitutive promoter: 
            pPP2AA3-PhiC31-tUBQ1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7643)
  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 7643)
  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: L1P1
FEATURES             Location/Qualifiers
     source          1..7643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    241..263
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    271..447
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(679..1704)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(1724..2069)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1761..1781)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2272..2294
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2286..2303
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2287..2303
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2310..2329
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2310..2328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2911..2922
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2970..4787
                     /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"
     misc_feature    5342..5366
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5457..6045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5537..5556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6216..7031)
                     /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(6216..7031)
                     /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     6939..6958
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7023..7042
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(7322..7643,1..114)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttcaattat gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta
       61 cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc
      121 ccacaacccg gcggccggcc gcaacagatc gttttataaa tttttttttt tgaaaaagaa
      181 aaagcccgaa aggcggcaac ctctcgggct tctggatttc cgatccccgg aattagatct
      241 tggcaggata tattgtggtg taacgttatc gatctggatt ttagtactgg attttggttt
      301 taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat actaagggtt
      361 tcttatatgc tcaacacatg agcgaaaccc tataagaacc ctaatttccc ttatcgggaa
      421 actactcaca cattatttat ggagaaaata gagagagata gatttgtaga gagagactgg
      481 tgatttcagc gtaccgaatt aattcctgaa tcactgcgac cggccctccc gcgacccagc
      541 cgagcgagct tagcgaactg tggacgagaa ctgtgccacc aagcgtaagg ccgttctctc
      601 gcatttgcct tgctaggctc gcgcgagttg ctggctgagg cgttctcgaa atcagctctt
      661 gttcggtcgg catctactct attcctttgc cctcggacga gtgctggggc gtcggtttcc
      721 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
      781 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
      841 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
      901 gcggagcata tacgcccgga gccttggcga tcctgcaagc tccggatgcc tccgctcgaa
      961 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
     1021 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
     1081 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
     1141 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
     1201 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
     1261 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
     1321 gctaagatcg gccgcagcga tagcatccat agcctccgcg accggctgaa gaacagcggg
     1381 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
     1441 ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
     1501 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
     1561 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
     1621 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt
     1681 cgcggtgagt tcaggctttt tcataggggg gatcagcttg ggctgtcctc tccaaatgaa
     1741 atgaacttcc ttatatagag gaagggtctt gcgaaggata gtgggattgt gcgtcatccc
     1801 ttacgtcagt ggagatgtca catcaatcca cttgctttga agacgtggtt ggaacgtctt
     1861 ctttttccac gatgctcctc gtgggtgggg gtccatcttt gggaccactg tcggcagagg
     1921 catcttgaat gatagccttt cctttatcgc aatgatggca tttgtaggag ccaccttcct
     1981 tttctactgt cctttcgatg aagtgacaga tagctgggca atggaatccg aggaggtttc
     2041 ccgaaattac cctttgttga aaagtctcaa tagccctttg gtcttctgag actgtatctt
     2101 tgacattttt ggagtagggg tacgataaca ttaacgctta caatttccat tcgccattca
     2161 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     2221 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     2281 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc ttggaggagg
     2341 aagaagaaga gagttactga gaagcttcct ttatacgaga gagatattaa acgaagtcgt
     2401 atttgttgga tatatgggcc tttatgtatt atacataagc ccatttagtt cttatctttg
     2461 acaggcccaa ttatatggac acagcttgaa gatggtttag ctgctgcgaa agacgagctc
     2521 cggtctgatt tctcgttctt ctgattgata gatcgctcgg aacttggaaa gcagcgtaat
     2581 cggtaaattc tcgatcattt cttcaatttc gaaacatttt ccaattgtat attgagaatc
     2641 tcaaatcgaa ttattgttta gatcttctct ctttattagt ctaagaatgg taatagtttc
     2701 tctctagatt gggacggcac aaagtttcag ttctccattc taatcatttt ttagaggcta
     2761 atgggtcacc tttgaattat ctcagaggcg gaattgagtt taaagtttgt aactttatcg
     2821 tcttttgtat tgatggtgtc tatggcaaag gtgattaact gcaaaggtat gcattgtgtt
     2881 ttattctgac tttgtgtaca tttgtgtttg attgaaggta gggagtggca aaggtgatta
     2941 actgcaaagg atttgagttt tggtgaggaa tggacacgta tgccggcgct tatgatagac
     3001 aatcaagaga aagagagaac tcttctgccg cctctccagc tacacagaga tctgccaacg
     3061 aggataaagc tgctgattta caaagagagg tagagagaga tgggggtagg tttagatttg
     3121 tgggtcattt ctctgaggct cctggtacat cagcctttgg tactgctgaa cggcctgaat
     3181 ttgaaagaat attaaatgag tgtcgggccg gtcgactaaa tatgataata gtttatgatg
     3241 tatctaggtt cagtagactc aaagttatgg atgcaattcc gatcgtgagc gagctacttg
     3301 cactcggtgt tactatcgta agtacacaag aaggtgtatt caggcagggt aatgtgatgg
     3361 accttataca ccttattatg cgactcgacg ctagccataa agagtcatca ctgaagagtg
     3421 ctaaaatctt ggacaccaag aacctgcaaa gggagcttgg aggctatgta ggaggaaaag
     3481 ctccgtatgg tttcgaactc gtgagtgaga caaaagagat taccagaaat ggtaggatgg
     3541 tgaacgtggt tattaataag ctagctcata gcacaacccc tctcacaggg ccttttgagt
     3601 tcgaacctga cgttattaga tggtggtgga gagagatcaa aactcacaaa catcttccat
     3661 ttaagccagg tagccaggct gctatccacc ctggatctat cactgggctt tgtaaaagga
     3721 tggatgctga cgcagtgcca actcgagggg aaacaatcgg taagaagact gcatcttccg
     3781 cttgggatcc cgctacagtt atgagaatcc ttcgggatcc gcgtattgct ggattcgccg
     3841 cagaagtcat ttataagaaa aaacctgatg gtactcctac tacaaaaatc gaaggttata
     3901 ggatccaaag ggatcctatt actttgcgtc cagttgaatt agactgtgga ccgatcatcg
     3961 aacctgccga gtggtacgag ttgcaggcct ggcttgatgg tagagggcgt ggaaagggtt
     4021 tatctcgggg acaggcaatt ttgtcagcaa tggataagct ttattgtgag tgtggggctg
     4081 ttatgacatc aaaacgtgga gaagagtcaa tcaaggattc ttacaggtgc agacgtcgta
     4141 aagtggtgga cccttctgct ccaggacagc atgagggtac atgcaacgtt tctatggctg
     4201 ctcttgataa atttgtggcc gaacgtattt tcaacaagat tagacacgct gaaggggatg
     4261 aggaaacttt agctcttctt tgggaggctg cccggcgttt tggtaagcta acagaggcac
     4321 cagaaaaaag tggcgaaaga gctaatttgg tagccgaaag agcagacgcc ttgaatgcat
     4381 tggaagaact ttacgaagat agggctgccg gagcatatga tggaccagtt ggtcggaagc
     4441 atttcaggaa gcaacaagct gcactaactt tgagacagca aggcgcagag gagcgactcg
     4501 cagagcttga ggccgccgag gccccaaagc ttccactcga tcaatggttt cctgaggatg
     4561 ccgacgctga tcctacaggt ccaaagagct ggtggggaag agctagcgta gacgataaga
     4621 gagtgtttgt gggtctattt gttgacaaga ttgtagttac taagtccact acgggtcgag
     4681 gacagggcac tcccatcgaa aagagagcca gtattacctg ggcaaagcct ccaaccgatg
     4741 atgatgaaga cgatgcacaa gatggtaccg aggatgtggc agcataataa gcttagagac
     4801 tcttatcaag aatcccatct cttgcttgct tttttttgtt gtcttccctt tgatagggtt
     4861 tgtttttctt gtttcagtga ctttctatgt taaaagataa tgtcagtaaa aggatttggt
     4921 tttctattat tctgaatcga ttacggaaga ttcttgctta attccaatct atacaagtat
     4981 cgtgaaataa tgaccgttta tgtgattagg agacgtgttt cattaataaa atataagatc
     5041 aatacattgt tagtagtgat aaactatgta caaattgtat tgattgtaaa agaaacacaa
     5101 taggttcctt ttttctacaa tatattgtga cagactctct gttttaacga atgaattaaa
     5161 tttgtcaacg aaaaaattaa agaatgatgt gatcactttg caccttggtt catcttcaga
     5221 ggcttctgca tgttccatgc attgaggctc tctacggtca caggttgtga gccgttgtta
     5281 aagacaaaca aatgcgctac cacagaatca ggggataacg caggaaagaa catgaaggcc
     5341 ttgacaggat atattggcgg gtaaactaag tcgctgtatg tgtttgtttg agatctcatg
     5401 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5461 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5521 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5581 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5641 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5701 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5761 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5821 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5881 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5941 ggaagaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6001 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6061 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6121 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6181 atctaaagta tatatgtgta acattggtct agtgattaga aaaactcatc gagcatcaaa
     6241 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     6301 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     6361 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     6421 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     6481 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     6541 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     6601 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     6661 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     6721 ttccctggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     6781 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     6841 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     6901 tacaatcggt agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     6961 tataaatcag catccatgtt ggaatttaat cgcggccttg agcaagacgt ttcccgttga
     7021 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     7081 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     7141 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     7201 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     7261 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcgatc
     7321 cccatccaac agcccgccgt cgagcgggct tttttatccc cggaagcctg tggatagagg
     7381 gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg ggctttccgg
     7441 cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat cgctaatcgg
     7501 agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc acgtgagaac
     7561 gctaatagcc ctttcagatc aacagcttgc aaacacccct cgctccggca agtagttaca
     7621 gcaagtagta tgttcaatta gct
//
