LOCUS       sequence-331569-        3311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phytobrick - AtHSP18.2t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3311)
  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 3311)
  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: pUPD+AtHSP18.2t
FEATURES             Location/Qualifiers
     source          1..3311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      10..258
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        complement(361..379)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(362..379)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(397..413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(397..413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(410..432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    421..437
                     /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(445..451,452..469,470..475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    490..511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(628..645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(799..1387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(879..898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1558..2349,2350..2418))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2181..2200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2419..2523)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2601..3056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2738..2759)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2950..2969
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3182..3204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3196..3213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3197..3213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3220..3239
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3220..3238
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3269..3278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3273..3282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcggcttat atgaagatga agatgaaata tttggtgtgt caaataaaaa gcttgtgtgc
       61 ttaagtttgt gtttttttct tggcttgttg tgttatgaat ttgtggcttt ttctaatatt
      121 aaatgaatgt aagatctcat tataatgaat aaacaaatgt ttctataatc cattgtgaat
      181 gttttgttgg atctcttctg cagcatataa ctactgtatg tgctatggta tggactatgg
      241 aatatgatta aagataagcg ctcgagacca ctcatcgccc aatcactagt gaattcgcgg
      301 ccgcctgcag gtcgaccata tgggagagct cccaacgcgt tggatgcata gcttgagtat
      361 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      421 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      481 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      541 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      601 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      661 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      721 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      781 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      841 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      901 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      961 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1021 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1081 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1141 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1201 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1261 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1321 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1381 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1441 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1501 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1561 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1621 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1681 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1741 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1801 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1861 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1921 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1981 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2041 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2101 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2161 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2221 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2281 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2341 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2401 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2461 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2521 gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa
     2581 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2641 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2701 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2761 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2821 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2881 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2941 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3001 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     3061 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3121 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3181 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     3241 gaattgggcc cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgattggg
     3301 cgatgagtgg t
//
