LOCUS       sequence-304703-        5464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for STARR-seq in tobacco leaves. Enhancer candidates can be 
            inserted just upstream of the minimal promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5464)
  AUTHORS   Jores T, Tonnies J, Dorrity MW, Cuperus J, Fields S, Queitsch C
  TITLE     Identification of Plant Enhancers and Their Constituent Elements by 
            STARR-seq in Tobacco Leaves.
  JOURNAL   Plant Cell. 2020 May 14. pii: tpc.20.00155. doi: 
            10.1105/tpc.20.00155.
  PUBMED    32409318
REFERENCE   2  (bases 1 to 5464)
  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: "Plant Cell.
            2020 May 14. pii: tpc.20.00155. doi: 10.1105/tpc.20.00155."
COMMENT     Sequence Label: pPSup
FEATURES             Location/Qualifiers
     source          1..5464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     70..86
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     71..87
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(183..185,186..899)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(225..246)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(486..505)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     833..854
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1220..1240)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1220..1238)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1259..1275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1259..1275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1272..1294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1283..1299
                     /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(1307..1313,1314..1331,1332..1337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1352..1373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1490..1507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1612..1636
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1727..2315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1807..1826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2486..3274)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNGTLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3565..4000
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4128..4150
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(4174..4426)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4446..4997)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(5039..5218)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     join(5450..5464,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5464,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
       61 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattccac tccgagacca
      121 caaggcgcgc ctagtggtct ccaggagctg ccgtcttcag tgcctgcagg catagaagac
      181 ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      241 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      301 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      361 tcgtgaccac cttcacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      421 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      481 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      541 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      601 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
      661 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
      721 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      781 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      841 tgctggagtt cgtgaccgcc gccgggatca ctcacggcat ggacgagctg tacaagtaaa
      901 gcggccgcaa attgcccaga attaattccc caactgcaca gaacaaattt ccctttgctt
      961 ttgtgtagac ctcaaaactt tatcccccat ctttgatctt atcccttgtt tttctgcttt
     1021 tttcttcttt cttgggtttt gatttccgga cttaacgttt gttttccggt ttgcgagaca
     1081 tattctatcg gattctcaac tgtctgatga aataaatatg taatgttcta taagtctttc
     1141 aatttgatgg gatgttccac ttctgtatag gagccaattg attgacaaca tgcatcaatc
     1201 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     1261 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1321 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1441 gccaacgctc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1501 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1561 tacggttatc cacagaatca ggggataacg caggaaagaa catgaaggcc ttgacaggat
     1621 atattggcgg gtaaactaag tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag
     1681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaagaagag
     2221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatt agattatcaa
     2401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2461 tatatgtgta acattggtct agtgattatt tgccgactac cttggtgatc tcgcctttca
     2521 cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttgtccaa
     2581 gataagcctg cctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
     2641 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
     2701 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata
     2761 gcgttaaggt tccatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
     2821 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
     2881 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc
     2941 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
     3001 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
     3061 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
     3121 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
     3181 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
     3241 gagtcgatac ttcggcgatc accgcttccc tcataacacc ccttgtatta ctgtttatgt
     3301 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     3361 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     3421 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     3481 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     3541 atgatggggc gattcaggcg atccccatcc aacagcccgc cgtcgagcgg gcttttttat
     3601 ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag
     3661 ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca
     3721 gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc
     3781 taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc
     3841 cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat
     3901 atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc
     3961 gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg
     4021 ccggccgcaa cagatcgttt tataaatttt ttttttttga aaaagaaaaa gcccgaaagg
     4081 cggcaacctc tcgggcttct ggatttccga tccccggaat tagatcttgg caggatatat
     4141 tgtggtgtaa cgttatcagc ttgcatgccg gtcgatctag taacatagat gacaccgcgc
     4201 gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata
     4261 attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa
     4321 ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa
     4381 caggattcaa tcttaagaaa ctttattgcc aaatgtttta acgatctgct tgactctagg
     4441 ggtcatcaga tttcggtgac gggcaggacc ggacggggcg gcaccggcag gctgaagtcc
     4501 agctgccaga aacccacgtc atgccagttc ccgtgcttga agccggccgc ccgcagcatg
     4561 ccgcgggggg catatccgag cgcctcgtgc atgcgcacgc tcgggtcgtt gggcagcccg
     4621 atgacagcga ccacgctctt gaagccctgt gcctccaggg acttcagcag gtgggtgtag
     4681 agcgtggagc ccagtcccgt ccgctggtgg cggggggaga cgtacacggt ggactcggcc
     4741 gtccagtcgt aggcgttgcg tgccttccag ggacccgcgt aggcgatgcc ggcgacctcg
     4801 ccgtccacct cggcgacgag ccagggatag cgctcccgca gacggacgag gtcgtccgtc
     4861 cactcctgcg gttcctgcgg ctcggtacgg aagttgaccg tgcttgtctc gatgtagtgg
     4921 ttgacgatgg tgcagaccgc cggcatgtcc gcctcggtgg cacggcggat gtcggccggg
     4981 cgtcgttctg ggctcatggt agatcccctc gatcgagttg agagtgaata tgagactcta
     5041 attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa gaaatatttg
     5101 ctagcttata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     5161 cttagctcat taaactccag aaacccgcgg ctcagtggct ccttcaacgt tgcggttctg
     5221 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     5281 cttaattctc cgctcatgat cgataacatt aacgcttaca atttccattc gccattcagg
     5341 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     5401 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     5461 cgtt
//
