LOCUS       sequence-304713-        5653 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. The GFP reporter 
            gene harbors the IV2 intron..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  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 5653)
  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_iGFP
FEATURES             Location/Qualifiers
     source          1..5653
                     /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"
     primer_bind     complement(225..246)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          286..474
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     complement(675..694)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1022..1043
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1409..1429)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1409..1427)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1448..1464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1448..1464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1461..1483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1472..1488
                     /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(1496..1502,1503..1520,1521..1526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1541..1562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1679..1696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1801..1825
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1916..2504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1996..2015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2675..3463)
                     /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      3754..4189
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4317..4339
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(4363..4615)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4635..5186)
                     /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(5228..5407)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     join(5639..5653,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5653,1..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 cgaaggtaag tttctgcttc
      301 tacctttgat atatatataa taattatcat taattagtag taatataata tttcaaatat
      361 ttttttcaaa ataaaagaat gtagtatata gcaattgctt ttctgtagtt tataagtgtg
      421 tatattttaa tttataactt ttctaatata tgaccaaaat ttgttgatgt gcaggtgatg
      481 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      541 ggcccaccct cgtgaccacc ttcacctacg gcgtgcagtg cttcagccgc taccccgacc
      601 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      661 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      721 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      781 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      841 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      901 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      961 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1021 acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg gacgagctgt
     1081 acaagtaaag cggccgcaaa ttgcccagaa ttaattcccc aactgcacag aacaaatttc
     1141 cctttgcttt tgtgtagacc tcaaaacttt atcccccatc tttgatctta tcccttgttt
     1201 ttctgctttt ttcttctttc ttgggttttg atttccggac ttaacgtttg ttttccggtt
     1261 tgcgagacat attctatcgg attctcaact gtctgatgaa ataaatatgt aatgttctat
     1321 aagtctttca atttgatggg atgttccact tctgtatagg agccaattga ttgacaacat
     1381 gcatcaatcg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     1441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1621 aatgaatcgg ccaacgctcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgaaggcct
     1801 tgacaggata tattggcggg taaactaagt cgctgtatgt gtttgtttga gatctcatgt
     1861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2401 gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatta
     2581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2641 tctaaagtat atatgtgtaa cattggtcta gtgattattt gccgactacc ttggtgatct
     2701 cgcctttcac gtagtgaaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt
     2761 cttgtccaag ataagcctgc ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
     2821 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
     2881 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
     2941 agttccatag cgttaaggtt ccatttagcg cctcaaatag atcctgttca ggaaccggat
     3001 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
     3061 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
     3121 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
     3181 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
     3241 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
     3301 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
     3361 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
     3421 ctgatagttg agtcgatact tcggcgatca ccgcttccct cataacaccc cttgtattac
     3481 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     3541 aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt
     3601 tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt
     3661 tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt
     3721 tctggctgga tgatggggcg attcaggcga tccccatcca acagcccgcc gtcgagcggg
     3781 cttttttatc cccggaagcc tgtggataga gggtagttat ccacgtgaaa ccgctaatgc
     3841 cccgcaaagc cttgattcac ggggctttcc ggcccgctcc aaaaactatc cacgtgaaat
     3901 cgctaatcag ggtacgtgaa atcgctaatc ggagtacgtg aaatcgctaa taaggtcacg
     3961 tgaaatcgct aatcaaaaag gcacgtgaga acgctaatag ccctttcaga tcaacagctt
     4021 gcaaacaccc ctcgctccgg caagtagtta cagcaagtag tatgttcaat tagcttttca
     4081 attatgaata tatatatcaa ttattggtcg cccttggctt gtggacaatg cgctacgcgc
     4141 accggctccg cccgtggaca accgcaagcg gttgcccacc gtcgagcgcc tttgcccaca
     4201 acccggcggc cggccgcaac agatcgtttt ataaattttt tttttttgaa aaagaaaaag
     4261 cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt agatcttggc
     4321 aggatatatt gtggtgtaac gttatcagct tgcatgccgg tcgatctagt aacatagatg
     4381 acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt
     4441 aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt
     4501 acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa
     4561 gaccggcaac aggattcaat cttaagaaac tttattgcca aatgttttaa cgatctgctt
     4621 gactctaggg gtcatcagat ttcggtgacg ggcaggaccg gacggggcgg caccggcagg
     4681 ctgaagtcca gctgccagaa acccacgtca tgccagttcc cgtgcttgaa gccggccgcc
     4741 cgcagcatgc cgcggggggc atatccgagc gcctcgtgca tgcgcacgct cgggtcgttg
     4801 ggcagcccga tgacagcgac cacgctcttg aagccctgtg cctccaggga cttcagcagg
     4861 tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc ggggggagac gtacacggtg
     4921 gactcggccg tccagtcgta ggcgttgcgt gccttccagg gacccgcgta ggcgatgccg
     4981 gcgacctcgc cgtccacctc ggcgacgagc cagggatagc gctcccgcag acggacgagg
     5041 tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga agttgaccgt gcttgtctcg
     5101 atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg cctcggtggc acggcggatg
     5161 tcggccgggc gtcgttctgg gctcatggta gatcccctcg atcgagttga gagtgaatat
     5221 gagactctaa ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag
     5281 aaatatttgc tagcttatag tgaccttagg cgacttttga acgcgcaata atggtttctg
     5341 acgtatgtgc ttagctcatt aaactccaga aacccgcggc tcagtggctc cttcaacgtt
     5401 gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa
     5461 cgtgactccc ttaattctcc gctcatgatc gataacatta acgcttacaa tttccattcg
     5521 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5581 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5641 cagtcacgac gtt
//
