LOCUS       sequence-386327-        8673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 Intein(DnaE-Ssp)-PhiC31 cassette pUBQ10-IP-tUBQ1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8673)
  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 8673)
  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: L1P60
FEATURES             Location/Qualifiers
     source          1..8673
                     /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     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"
     promoter        3342..3687
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             4771..4791
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4829..5003
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5050..5074
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5165..5753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5245..5264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7262..8077)
                     /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(7262..8077)
                     /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     7985..8004
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8069..8088
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(8368..8673,1..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 aggaattcca atcccacaaa aatctgagct taacagcaca gttgctcctc
     2401 tcagagcaga atcgggtatt caacaccctc atatcaacta ctacgttgtg tataacggtc
     2461 cacatgccgg tatatacgat gactggggtt gtacaaaggc ggcaacaaac ggcgttcccg
     2521 gagttgcaca caagaaattt gccactatta cagaggcaag agcagcagct gacgcgtaca
     2581 caacaagtca gcaaacagac aggttgaact tcatccccaa aggagaagct caactcaagc
     2641 ccaagagctt tgctaaggcc ctaacaagcc caccaaagca aaaagcccac tggctcacgc
     2701 taggaaccaa aaggcccagc agtgatccag ccccaaaaga gatctccttt gccccggaga
     2761 ttacaatgga cgatttcctc tatctttacg atctaggaag gaagttcgaa ggtgaaggtg
     2821 acgacactat gttcaccact gataatgaga aggttagcct cttcaatttc agaaagaatg
     2881 ctgacccaca gatggttaga gaggcctacg cagcaagtct catcaagacg atctacccga
     2941 gtaacaatct ccaggagatc aaataccttc ccaagaaggt taaagatgca gtcaaaagat
     3001 tcaggactaa ttgcatcaag aacacagaga aagacatatt tctcaagatc agaagtacta
     3061 ttccagtatg gacgattcaa ggcttgcttc ataaaccaag gcaagtaata gagattggag
     3121 tctctaaaaa ggtagttcct actgaatcta aggccatgca tggagtctaa gattcaaatc
     3181 gaggatctaa cagaactcgc cgtcaagact ggcgaacagt tcatacagag tcttttacga
     3241 ctcaatgaca agaagaaaat cttcgtcaac atggtggagc acgacactct ggtctactcc
     3301 aaaaatgtca aagatacagt ctcagaagat caaagggcta ttgagacttt tcaacaaagg
     3361 ataatttcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcgaaagg
     3421 acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggctatc
     3481 attcaagatc tctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
     3541 gtggaaaaag aagaggttcc aaccacgtct acaaagcaag tggattgatg tgacatctcc
     3601 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
     3661 ggaagttcat ttcatttgga gaggacacgc tcgagtataa gagctcattt ttacaacaat
     3721 taccaacaac aacaaacaac aaacaacatt acaattacat ttacaattat cgatacaatg
     3781 gtgaaagtga ttggccgccg cagcctgggc gtgcagcgca tttttgatat tggcctgcgc
     3841 caggatcata actttctgct ggcgaacggc gcgattgcgg cgaactgctt taacagggat
     3901 cctattactt tgcgtccagt tgaattagac tgtggaccga tcatcgaacc tgccgagtgg
     3961 tacgagttgc aggcctggct tgatggtaga gggcgtggaa agggtttatc tcggggacag
     4021 gcaattttgt cagcaatgga taagctttat tgtgagtgtg gggctgttat gacatcaaaa
     4081 cgtggagaag agtcaatcaa ggattcttac aggtgcagac gtcgtaaagt ggtggaccct
     4141 tctgctccag gacagcatga gggtacatgc aacgtttcta tggctgctct tgataaattt
     4201 gtggccgaac gtattttcaa caagattaga cacgctgaag gggatgagga aactttagct
     4261 cttctttggg aggctgcccg gcgttttggt aagctaacag aggcaccaga aaaaagtggc
     4321 gaaagagcta atttggtagc cgaaagagca gacgccttga atgcattgga agaactttac
     4381 gaagataggg ctgccggagc atatgatgga ccagttggtc ggaagcattt caggaagcaa
     4441 caagctgcac taactttgag acagcaaggc gcagaggagc gactcgcaga gcttgaggcc
     4501 gccgaggccc caaagcttcc actcgatcaa tggtttcctg aggatgccga cgctgatcct
     4561 acaggtccaa agagctggtg gggaagagct agcgtagacg ataagagagt gtttgtgggt
     4621 ctatttgttg acaagattgt agttactaag tccactacgg gtcgaggaca gggcactccc
     4681 atcgaaaaga gagccagtat tacctgggca aagcctccaa ccgatgatga tgaagacgat
     4741 gcacaagatg gtaccgagga tgtggcagca ccaaaaaaaa agaggaaggt gtaagcttct
     4801 ctagctagag tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag
     4861 ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt
     4921 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
     4981 aatccagtac taaaatccag atcgctacca cagaatcagg ggataacgca ggaaagaaca
     5041 tgaaggcctt gacaggatat attggcgggt aaactaagtc gctgtatgtg tttgtttgag
     5101 atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5161 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5221 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5281 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5341 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5401 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5461 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5521 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5581 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5641 ttaccttcgg aagaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5701 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5761 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5821 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5881 ttaaatcaat ctaaagtact gagagatccc ctcataattt ccccaaagcg taaccatgtg
     5941 tgaataaatt ttgagctagt agggttgcag ccacgagtaa gtcttccctt gttattgtgt
     6001 agccagaatg ccgcaaaact tccatgccta agcgaactgt tgagagtacg tttcgatttc
     6061 tgactgtgtt agcctggaag tgcttgtccc aaccttgttt ctgagcatga acgcccgcaa
     6121 gccaacatgt tagttgaagc atcagggcga ttagcagcat gatatcaaaa cgctctgagc
     6181 tgctcgttcg gctatggcgt aggcctagtc cgtaggcagg acttttcaag tctcggaagg
     6241 tttcttcaat ctgcattcgc ttcgaataga tattaacaag ttgtttgggt gttcgaattt
     6301 caacaggtaa gttagttgct agaacccatg gctcctttgc cgacgctgag tagattttag
     6361 gtgacgggtg gtgacaatga gtccgtgtcg agcgctgatt ttttcggcct ttagagcgag
     6421 atttatacaa tagaatttgg catgagattg gattgctttt agtcagcctc ttatagccta
     6481 aagtctttga gtgactagat gacatatcat gtaagttgct gataggtttc cagttttccg
     6541 ctcctaggtc tgcatattgt acttttcctc ttactcgact taaccagtac caacccagct
     6601 tctcaacgga tttataccat ggcactttaa agccagcatc actgacaatg agcggtgtgg
     6661 tgttactcgg tagaatgctc gcaaggtcgg ctagaaattg gtcatgagct ttctttgaac
     6721 attgctctga aagcgggaac gctttctcat aaagagtaac agaacgaccg tgtagtgcga
     6781 ctgaagctcg caataccata agtcgttttt gctcacgaat atcagaccag tcaacaagta
     6841 caatgggcat cgtattgccc gaacagataa agctagcatg ccaacggtat acagcgagtc
     6901 gctctttgtg gaggtgacga ttacctaaca atcggtcgat tcgtttgatg ttatgttttg
     6961 ttctcgcttt ggttggcagg ttacggccaa gttcggtaag agtgagagtt ttacagtcaa
     7021 gtaatgcgtg gcaagccaac gttaagctgt tgagtcgttt taagtgtaat tcggggcaga
     7081 attggtaaag agagtcgtgt aaaatatcga gttcgcacat cttgttgtct gattattgat
     7141 ttttcgcgaa accatttgat catatgacaa gatgtgtatc caccttaact taatgatttt
     7201 taccaaaatc attaggggat tcatcagtct aaagtatata tgtgtaacat tggtctagtg
     7261 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     7321 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     7381 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     7441 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     7501 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     7561 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     7621 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     7681 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     7741 attcttctaa tacctggaat gctgttttcc ctgggatcgc agtggtgagt aaccatgcat
     7801 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     7861 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     7921 acaactctgg cgcatcgggc ttcccataca atcggtagat tgtcgcacct gattgcccga
     7981 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     8041 gccttgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     8101 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     8161 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     8221 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     8281 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     8341 tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag cgggcttttt
     8401 tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta atgccccgca
     8461 aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa
     8521 tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat
     8581 cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca gcttgcaaac
     8641 acccctcgct ccggcaagta gttacagcaa gta
//
