LOCUS       sequence-386881-        6571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 inducibile estradiol promter and XVE activator protein 
            (p35S-XVE gene-tRBCS-tNOS-LexA-p35S with GG0 and GG1 golden gate 
            linkers).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6571)
  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 6571)
  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: L0-I35 p35S-XVE-pEstradiol GG0-GG1
FEATURES             Location/Qualifiers
     source          1..6571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..471
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             502..753
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA BD
                     /note="binds the 16-bp palindromic sequence 
                     5'-CTGTATATATATACAG-3'"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     CDS             769..1002
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1012..1947
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             1060..1953
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      2397..2648
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2969..3015
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(3172..3188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3172..3188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3185..3207)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3196..3212
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3220..3226,3227..3244,3245..3250))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3265..3286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3403..3420)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3574..4162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3654..3673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4300..4674)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      4671..4680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4880..5674)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4880..5674)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4991..5010)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5601..5620
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(6023..6325)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(6125..6144)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     6448..6470
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6462..6479
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6463..6479
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6486..6505
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6486..6504
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactctgtgg tctcaggagc cacgataccg gtggtaccca tgcctgcagg tcaacatggt
       61 ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag aagaccagag
      121 ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc
      181 agctatctgt cacttcatcg aaaggacagt agaaaaggaa gatggcttct acaaatgcca
      241 tcattgcgat aaaggaaagg ctatcgttca agatgcctct accgacagtg gtcccaaaga
      301 tggaccccca cccacgagga acatcgtgga aaaagaagac gttccaacca cgtcttcaaa
      361 gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc
      421 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga cctcctcgag
      481 tacgtaccaa ttccgggcgg aatgaaagcg ttaacggcca ggcaacaaga ggtgtttgat
      541 ctcatccgtg atcacatcag ccagacaggt atgccgccga cgcgtgcgga aatcgcgcag
      601 cgtttggggt tccgttcccc aaacgcggct gaagaacatc tgaaggcgct ggcacgcaaa
      661 ggcgttattg aaattgtttc cggcgcatca cgcgggattc gtctgttgca ggaagaggaa
      721 gaagggttgc cgctggtagg tcgtgtggct gccggtgaac cgtcgagcgc ccccccgacc
      781 gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc
      841 gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga
      901 tttacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag
      961 cagatgttta ccgatgccct tggaattgac gagtacggtg gggatccgtc tgctggagac
     1021 atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc
     1081 ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc
     1141 atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta
     1201 ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca
     1261 ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag
     1321 atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagggaagct actgtttgct
     1381 cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc
     1441 gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt
     1501 gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc
     1561 ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact
     1621 ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc
     1681 cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg
     1741 tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga gatgctggac
     1801 gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa
     1861 agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg
     1921 ggggaggcag agggtttccc tgccacagtc tgagagctcc ctggcggaat tcccagagat
     1981 gttagctgaa atcatcacta atcagatacc aaaatattca aatggaaata tcaaaaagct
     2041 tctgtttcat caaaaatgac tcgacctaac tgagtaagct agcttgttcg agtattatgg
     2101 cattgggaaa actgtttttc ttgtaccatt tgttgtgctt gtaatttact gtgtttttta
     2161 ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg agaagagtta atgaatgata
     2221 tggtcctttt gttcattctc aaattaatat tatttgtttt ttctcttatt tgttgtgtgt
     2281 tgaatttgaa attataagag atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc
     2341 gtggcctcta atgaccgaag ttaatatgag gagtaaaaca tcccaaacgc ggccgcatcg
     2401 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     2461 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     2521 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     2581 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     2641 actagatcgg gaattgatcc cccctcgaca gcttgcatgc cgcttgggct gcaggtcgag
     2701 gctaaaaaac taatcgcatt atcatcccct cgacgtactg tacatataac cactggtttt
     2761 atatacagca gtactgtaca tataaccact ggttttatat acagcagtcg acgtactgta
     2821 catataacca ctggttttat atacagcagt actgtacata taaccactgg ttttatatac
     2881 agcagtcgag gtaagattag atatggatat gtatatggat atgtatatgg tggtaatgcc
     2941 atgtaatatg ctcgactcta ggatcttcgc aagacccttc ctctatataa ggaagttcat
     3001 ttcatttgga gaggacacgc tgaagctagt cgactctagc aatgtgagac catcaacgtc
     3061 ctgaattcca gcacactggc ggccgttact agtggatccg agctcggtac caagcttgat
     3121 gcatagcttg agtattctaa cgcgtcacct aaatagcttg gcgtaatcat ggtcatagct
     3181 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3241 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3301 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3361 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3421 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3481 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3541 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3601 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3661 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3721 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3781 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3841 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3901 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3961 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4021 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4081 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4141 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4201 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4261 ctagatcctt ttaaattaaa aatgaagttt tagcacgtgt cagtcctgct cctcggccac
     4321 gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc
     4381 gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga
     4441 ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg
     4501 caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc
     4561 gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc
     4621 tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatggtggc
     4681 cctcctcacg tgctattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4801 gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     4861 aattgtaagc gttaataatt cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     4921 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     4981 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     5041 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     5101 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg
     5161 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     5221 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     5281 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     5341 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     5401 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     5461 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     5521 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     5581 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     5641 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     5701 gatcagagct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     5761 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg
     5821 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     5881 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     5941 gtcagcaccg tttctgcgga ctggctttct acgtgaaaag gatctaggtg aagatccttt
     6001 ttgataatct catgcctgac atttatattc cccagaacat caggttaatg gcgtttttga
     6061 tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag accggcacac
     6121 tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc accgggtaaa
     6181 gttcacggga gactttatct gacagcagac gtgcactggc cagggggatc accatccgtc
     6241 gccccggcgt gtcaataata tcactctgta catccacaaa cagacgataa cggctctctc
     6301 ttttataggt gtaaacctta aactgccgta cgtataggct gcgcaactgt tgggaagggc
     6361 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     6421 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     6481 aattgtaata cgactcacta tagggcgaat tgggccctct agatgcatgc tcgagcggcc
     6541 gccagtgtga tggatatctg cagaattcag g
//
