LOCUS       sequence-386331-        9817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 PhiC31-intein(DnaE-Npu) cassette : p35S-PI-t35S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9817)
  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 9817)
  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: L1P59
FEATURES             Location/Qualifiers
     source          1..9817
                     /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             5619..5636
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6194..6218
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6309..6897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6389..6408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8406..9221)
                     /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(8406..9221)
                     /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     9129..9148
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9213..9232
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(9512..9817,1..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 agctcaacaa caaactttcc attcacccat ttatggatac acagtctagc
     2401 tcaacagagc ttttaaccca aattggtaca atagaataca actttagatc ataattctca
     2461 aaagaaagag attccttagc tattctatct gccactccat ttccttctcg gcttgtatgc
     2521 acaagcataa aatcctcaaa cttgctaagt agatacttta tgtcttggat aattggattg
     2581 agacttgaca agcataactt tcatgtaacc aaagacacaa gttgctgaga atccacctca
     2641 aaaatgatct tcctataatt gaatcgggat aatgacagca cagcccatct aagagcctcc
     2701 acttctactt ccagcacgct tcttactttt accacagctc ttgcacctaa ccataacacc
     2761 ttccctgtat gatcgcgaag cacccaccct aagccacatt ttaatccttc tgttggccat
     2821 gccccatcaa agttgcactt aacccaagat tgtggtggag cttcccatgt ttctcgtctg
     2881 tcccgacggt gttgtggttg gtgctttcct tacattctga gcctctttcc ttctaatcca
     2941 ctcatctgca tcttcttgtg tccttactaa tacctcattg gttccaaatt ccctcccttt
     3001 aagcaccagc tcgtttctgt tcttccacag cctcccaagt atccaaggga ctaaagcctc
     3061 cacattcttc agatcaggat attcttgttt aagatgttga actctatgga ggtttgtatg
     3121 aactgatgat ctaggaccgg ataagttccc ttcttcatag cgaacttatt caaagaatgt
     3181 tttgtgtatc attcttgtta cattgttatt aatgaaaaaa tattattggt cattggactg
     3241 aacacgagtg ttaaatatgg accaggcccc aaataagatc cattgatata tgaattaaat
     3301 aacaagaata aatcgagtca ccaaaccact tgcctttttt aacgagactt gttcaccaac
     3361 ttgatacaaa agtcattatc ctatgcaaat caataatcat acaaaaatat ccaataacac
     3421 taaaaaatta aaagaaatgg ataatttcac aatatgttat acgataaaga agttactttt
     3481 ccaagaaatt cactgatttt ataagcccac ttgcattaga taaatggcaa aaaaaaacaa
     3541 aaaggaaaag aaataaagca cgaagaattc tagaaaatac gaaatacgct tcaatgcagt
     3601 gggacccacg gttcaattat tgccaatttt cagctccacc gtatatttaa aaaataaaac
     3661 gataatgcta aaaaaatata aatcgtaacg atcgttaaat ctcaacggct ggatcttatg
     3721 acgaccgtta gaaattgtgg ttgtcgacga gtcagtaata aacggcgtca aagtggttgc
     3781 agccggcaca cacgagtcgt gtttatcaac tcaaagcaca aatacttttc ctcaacctaa
     3841 aaataaggca attagccaaa aacaactttg cgtgtaaaca acgctcaata cacgtgtcat
     3901 tttattatta gctattgctt caccgcctta gctttctcgt gacctagtcg tcctcgtctt
     3961 ttcttcttct tcttctataa aacaataccc aaagagctct tcttcttcac aattcagatt
     4021 tcaatttctc aaaatcttaa aaactttctc tcaattctct ctaccgtgat caaggtaaat
     4081 ttctgtgttc cttattctct caaaatcttc gattttgttt tcgttcgatc ccaatttcgt
     4141 atatgttctt tggtttagat tctgttaatc ttagatcgaa gacgattttc tgggtttgat
     4201 cgttagatat catcttaatt ctcgattagg gtttcataga tatcatccga tttgttcaaa
     4261 taatttgagt tttgtcgaat aattactctt cgatttgtga tttctatcta gatctggtgt
     4321 tagtttctag tttgtgcgat cgaatttgtc gattaatctg agtttttctg attaacagaa
     4381 tggacacgta tgccggcgct tatgatagac aatcaagaga aagagagaac tcttctgccg
     4441 cctctccagc tacacagaga tctgccaacg aggataaagc tgctgattta caaagagagg
     4501 tagagagaga tgggggtagg tttagatttg tgggtcattt ctctgaggct cctggtacat
     4561 cagcctttgg tactgctgaa cggcctgaat ttgaaagaat attaaatgag tgtcgggccg
     4621 gtcgactaaa tatgataata gtttatgatg tatctaggtt cagtagactc aaagttatgg
     4681 atgcaattcc gatcgtgagc gagctacttg cactcggtgt tactatcgta agtacacaag
     4741 aaggtgtatt caggcagggt aatgtgatgg accttataca ccttattatg cgactcgacg
     4801 ctagccataa agagtcatca ctgaagagtg ctaaaatctt ggacaccaag aacctgcaaa
     4861 gggagcttgg aggctatgta ggaggaaaag ctccgtatgg tttcgaactc gtgagtgaga
     4921 caaaagagat taccagaaat ggtaggatgg tgaacgtggt tattaataag ctagctcata
     4981 gcacaacccc tctcacaggg ccttttgagt tcgaacctga cgttattaga tggtggtgga
     5041 gagagatcaa aactcacaaa catcttccat ttaagccagg tagccaggct gctatccacc
     5101 ctggatctat cactgggctt tgtaaaagga tggatgctga cgcagtgcca actcgagggg
     5161 aaacaatcgg taagaagact gcatcttccg cttgggatcc cgctacagtt atgagaatcc
     5221 ttcgggatcc gcgtattgct ggattcgccg cagaagtcat ttataagaaa aaacctgatg
     5281 gtactcctac tacaaaaatc gaatattgcc tgagctatga aaccgaaatt ctgaccgtgg
     5341 aatatggcag cctgccgatt ggcaaaattg tggaaaaacg cattgaatgc accgtgtata
     5401 gcgtggataa caacggcaac atttataccc agccggtggc gcagtggcat gatcgcggcg
     5461 aacaggaagt gtttgaatat tgcctggaag atggcagcct gattcgcgcg accaaagatc
     5521 ataaatttat gaccgtggat ggccagatgc tgccgattga tgaaattttt gaacgcgaac
     5581 tggatctgat gcgcgtggat aacctgccga acaccagcca tcatcatcat catcattaat
     5641 aagcttagag actcttatca agaatcccat ctcttgcttg cttttttttg ttgtcttccc
     5701 tttgataggg tttgtttttc ttgtttcagt gactttctat gttaaaagat aatgtcagta
     5761 aaaggatttg gttttctatt attctgaatc gattacggaa gattcttgct taattccaat
     5821 ctatacaagt atcgtgaaat aatgaccgtt tatgtgatta ggagacgtgt ttcattaata
     5881 aaatataaga tcaatacatt gttagtagtg ataaactatg tacaaattgt attgattgta
     5941 aaagaaacac aataggttcc ttttttctac aatatattgt gacagactct ctgttttaac
     6001 gaatgaatta aatttgtcaa cgaaaaaatt aaagaatgat gtgatcactt tgcaccttgg
     6061 ttcatcttca gaggcttctg catgttccat gcattgaggc tctctacggt cacaggttgt
     6121 gagccgttgt taaagacaaa caaatgcgct accacagaat caggggataa cgcaggaaag
     6181 aacatgaagg ccttgacagg atatattggc gggtaaacta agtcgctgta tgtgtttgtt
     6241 tgagatctca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6481 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6721 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6781 ccagttacct tcggaagaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7021 agttttaaat caatctaaag tactgagaga tcccctcata atttccccaa agcgtaacca
     7081 tgtgtgaata aattttgagc tagtagggtt gcagccacga gtaagtcttc ccttgttatt
     7141 gtgtagccag aatgccgcaa aacttccatg cctaagcgaa ctgttgagag tacgtttcga
     7201 tttctgactg tgttagcctg gaagtgcttg tcccaacctt gtttctgagc atgaacgccc
     7261 gcaagccaac atgttagttg aagcatcagg gcgattagca gcatgatatc aaaacgctct
     7321 gagctgctcg ttcggctatg gcgtaggcct agtccgtagg caggactttt caagtctcgg
     7381 aaggtttctt caatctgcat tcgcttcgaa tagatattaa caagttgttt gggtgttcga
     7441 atttcaacag gtaagttagt tgctagaacc catggctcct ttgccgacgc tgagtagatt
     7501 ttaggtgacg ggtggtgaca atgagtccgt gtcgagcgct gattttttcg gcctttagag
     7561 cgagatttat acaatagaat ttggcatgag attggattgc ttttagtcag cctcttatag
     7621 cctaaagtct ttgagtgact agatgacata tcatgtaagt tgctgatagg tttccagttt
     7681 tccgctccta ggtctgcata ttgtactttt cctcttactc gacttaacca gtaccaaccc
     7741 agcttctcaa cggatttata ccatggcact ttaaagccag catcactgac aatgagcggt
     7801 gtggtgttac tcggtagaat gctcgcaagg tcggctagaa attggtcatg agctttcttt
     7861 gaacattgct ctgaaagcgg gaacgctttc tcataaagag taacagaacg accgtgtagt
     7921 gcgactgaag ctcgcaatac cataagtcgt ttttgctcac gaatatcaga ccagtcaaca
     7981 agtacaatgg gcatcgtatt gcccgaacag ataaagctag catgccaacg gtatacagcg
     8041 agtcgctctt tgtggaggtg acgattacct aacaatcggt cgattcgttt gatgttatgt
     8101 tttgttctcg ctttggttgg caggttacgg ccaagttcgg taagagtgag agttttacag
     8161 tcaagtaatg cgtggcaagc caacgttaag ctgttgagtc gttttaagtg taattcgggg
     8221 cagaattggt aaagagagtc gtgtaaaata tcgagttcgc acatcttgtt gtctgattat
     8281 tgatttttcg cgaaaccatt tgatcatatg acaagatgtg tatccacctt aacttaatga
     8341 tttttaccaa aatcattagg ggattcatca gtctaaagta tatatgtgta acattggtct
     8401 agtgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     8461 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     8521 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     8581 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     8641 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     8701 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     8761 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     8821 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     8881 ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt gagtaaccat
     8941 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     9001 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     9061 agaaacaact ctggcgcatc gggcttccca tacaatcggt agattgtcgc acctgattgc
     9121 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     9181 cgcggccttg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     9241 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     9301 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     9361 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     9421 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     9481 tggctggatg atggggcgat tcaggcgatc cccatccaac agcccgccgt cgagcgggct
     9541 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
     9601 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
     9661 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
     9721 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
     9781 aaacacccct cgctccggca agtagttaca gcaagta
//
