LOCUS       sequence-335134-        3287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phytobrick - 35St.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3287)
  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 3287)
  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+35St
FEATURES             Location/Qualifiers
     source          1..3287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(337..355)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(338..355)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(373..389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(373..389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(386..408)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    397..413
                     /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(421..427,428..445,446..451))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    466..487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(604..621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(775..1363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(855..874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1534..2325,2326..2394))
                     /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     2157..2176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2395..2499)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2577..3032)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2714..2735)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2926..2945
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3158..3180
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3172..3189
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3173..3189
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3196..3215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3196..3214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3245..3254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3249..3258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcggcttcg gccatgctag agtccgcaaa aatcaccagt ctctctctac aaatctatct
       61 ctctctattt ttctccagaa taatgtgtga gtagttccca gataagggaa ttagggttct
      121 tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt
      181 aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagtg acctcgctcg
      241 agaccactca tcgcccaatc actagtgaat tcgcggccgc ctgcaggtcg accatatggg
      301 agagctccca acgcgttgga tgcatagctt gagtattcta tagtgtcacc taaatagctt
      361 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
      421 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
      481 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
      541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
      601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1201 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1381 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1441 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1501 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1561 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1621 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     1681 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1741 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1801 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1861 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1921 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1981 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2041 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2101 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2161 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2221 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2281 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2341 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2401 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2461 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2521 acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2581 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2641 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2701 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2761 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2821 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2881 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2941 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3001 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     3061 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3121 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3181 acggccagtg aattgtaata cgactcacta tagggcgaat tgggcccgac gtcgcatgct
     3241 cccggccgcc atggcggccg cgggaattcg attgggcgat gagtggt
//
