LOCUS       sequence-331564-        4605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phytobrick - Estradiol Inducible Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4605)
  AUTHORS   Gonzalez-Grandio E, Demirer GS, Ma W, Brady S, Landry MP
  TITLE     A Ratiometric Dual Color Luciferase Reporter for Fast 
            Characterization of Transcriptional Regulatory Elements in Plants.
  JOURNAL   ACS Synth Biol. 2021 Sep 14. doi: 10.1021/acssynbio.1c00248.
  PUBMED    34520169
REFERENCE   2  (bases 1 to 4605)
  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: "ACS Synth 
            Biol. 2021 Sep 14. doi: 10.1021/acssynbio.1c00248."
COMMENT     Sequence Label: pUPD2+pEstrInd
FEATURES             Location/Qualifiers
     source          1..4605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        208..254
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             283..534
                     /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             550..783
                     /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             793..1728
                     /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             841..1734
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        2458..2504
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      2562..2619
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2643..2660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2814..3402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2894..2913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3585..3679)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3700..4359)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4274..4293
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4521..4539
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4543..4586)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(4589..4605,1..5)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tagagcgatg agtggtctcg ggagatagtt taaactgaag gcgggaaacg acaatctgat
       61 ccaagctcaa gctaagcttg catgcctgca ggatatcgtg gatccaagct tgccacgtgc
      121 cgccacgtgc cgccacgtgc cgccacgtgc ctctagagga tccatctcca ctgacgtaag
      181 ggatgacgca caatcccact atccttcgca agacccttcc tctatataag gaagttcatt
      241 tcatttggag aggacacgct gggatcccca attccgggcg gaatgaaagc gttaacggcc
      301 aggcaacaag aggtgtttga tctcatccgt gatcacatca gccagacagg tatgccgccg
      361 acgcgtgcgg aaatcgcgca gcgtttgggg ttccgttccc caaacgcggc tgaagaacat
      421 ctgaaggcgc tggcacgcaa aggcgttatt gaaattgttt ccggcgcatc acgcgggatt
      481 cgtctgttgc aggaagagga agaagggttg ccgctggtag gtcgtgtggc tgccggtgaa
      541 ccgtcgagcg cccccccgac cgatgtcagc ctgggggacg agctccactt agacggcgag
      601 gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg atctggacat gttgggggac
      661 ggggattccc cgggtccggg atttaccccc cacgactccg ccccctacgg cgctctggat
      721 atggccgact tcgagtttga gcagatgttt accgatgccc ttggaattga cgagtacggt
      781 ggggatccgt ctgctggaga catgagagct gccaaccttt ggccaagccc gctcatgatc
      841 aaacgctcta agaagaacag cctggccttg tccctgacgg ccgaccagat ggtcagtgcc
      901 ttgttggatg ctgagccccc catactctat tccgagtatg atcctaccag acccttcagt
      961 gaagcttcga tgatgggctt actgaccaac ctggcagaca gggagctggt tcacatgatc
     1021 aactgggcga agagggtgcc aggctttgtg gatttgaccc tccatgatca ggtccacctt
     1081 ctagaatgtg cctggctaga gatcctgatg attggtcttg tctggcgctc catggagcac
     1141 ccagggaagc tactgtttgc tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta
     1201 gagggcatgg tggagatctt cgacatgctg ctggctacat catctcggtt ccgcatgatg
     1261 aatctgcagg gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg
     1321 tacacatttc tgtccagcac cctgaagtct ctggaagaga aggaccatat ccaccgagtc
     1381 ctggacaaga tcacagacac tttgatccac ctgatggcca aggcaggcct gaccctgcag
     1441 cagcagcacc agcggctggc ccagctcctc ctcatcctct cccacatcag gcacatgagt
     1501 aacaaaggca tggagcatct gtacagcatg aagtgcaaga acgtggtgcc cctctatgac
     1561 ctgctgctgg agatgctgga cgcccaccgc ctacatgcgc ccactagccg tggaggggca
     1621 tccgtggagg agacagacca aagccacttg gccactgcgg gctctacttc atcgcattcc
     1681 ttgcaaaagt attacatcac gggggaggca gagggtttcc ctgccacagt ctgagagctc
     1741 cctggcggaa ttcccagaga tgttagctga aatcatcact aatcagatac caaaatattc
     1801 aaatggaaat atcaaaaagc ttctgtttca tcaaaaatga ctcgacctaa ctgagtaagc
     1861 tagcttgttc gagtattatg gcattgggaa aactgttttt cttgtaccat ttgttgtgct
     1921 tgtaatttac tgtgtttttt attcggtttt cgctatcgaa ctgtgaaatg gaaatggatg
     1981 gagaagagtt aatgaatgat atggtccttt tgttcattct caaattaata ttatttgttt
     2041 tttctcttat ttgttgtgtg ttgaatttga aattataaga gatatgcaaa cattttgttt
     2101 tgagtaaaaa tgtgtcaaat cgtggcctct aatgaccgaa gttaatatga ggagtaaaac
     2161 atcccaaaca gcttgggctg caggtcgagg ctaaaaaact aatcgcatta tcatcccctc
     2221 gacgtactgt acatataacc actggtttta tatacagcag tactgtacat ataaccactg
     2281 gttttatata cagcagtcga cgtactgtac atataaccac tggttttata tacagcagta
     2341 ctgtacatat aaccactggt tttatataca gcagtcgagg taagattaga tatggatatg
     2401 tatatggata tgtatatggt ggtaatgcca tgtaatatgc tcgactctag gatcttcgca
     2461 agacccttcc tctatataag gaagttcatt tcatttggag aggacacgct gaagctagtc
     2521 aatgtgagac cactcatcgc gactagtagc ggccgctgca gtccggcaaa aaagggcaag
     2581 gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct
     2641 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2701 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2761 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccac
     2821 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2881 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2941 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3001 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3061 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3121 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3181 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3241 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3301 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3361 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3421 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3481 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3541 taaagtatat atgagtaaac ttggtctgac agctcgaggc ttggattctc accaataaaa
     3601 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     3661 ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc cctgccactc
     3721 gtcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc
     3781 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
     3841 catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
     3901 gaaactcacc cagggattgg ctgatacgaa aaacatattc tcaataaacc ctttagggaa
     3961 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
     4021 gaaatcgtcg tggtattcac tccagagtga tgaaaacgtt tcagtttgct catggaaaac
     4081 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
     4141 aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
     4201 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
     4261 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
     4321 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
     4381 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     4441 ggaacctctt acgtgcccga tcaactcgag tgccacctga cgtctaagaa accattatta
     4501 tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat
     4561 ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttc
//
