LOCUS       sequence-304715-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for STARR-seq in tobacco leaves. Enhancer candidates can be 
            inserted into the IV2 intron in the GFP reporter gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5669)
  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 5669)
  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: pPSint
FEATURES             Location/Qualifiers
     source          1..5669
                     /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(199..220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(691..710)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1038..1059
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1425..1445)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1425..1443)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1464..1480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1464..1480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1477..1499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1488..1504
                     /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(1512..1518,1519..1536,1537..1542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1557..1578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1695..1712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1817..1841
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1932..2520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2012..2031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2691..3479)
                     /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      3770..4205
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4333..4355
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(4379..4631)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4651..5202)
                     /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(5244..5423)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     join(5655..5669,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5669,1..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 cgaattccgc ggccgcagga
      121 gctgccgtct tcagtgcctg caggcataga agacccgtga gcaagggcga ggagctgttc
      181 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      241 gtgtccggcg agggcgaagg taagtttctg cttctacctt tgatatatat ataataatta
      301 tcattaatta gtagtaatat aatatttcaa atattttttt caaaataaaa gaatgtagta
      361 tatagcaatt actccgagac cacaaggcgc gcctagtggt ctccaggaaa ttgcttttct
      421 gtagtttata agtgtgtata ttttaattta taacttttct aatatatgac caaaatttgt
      481 tgatgtgcag gtgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      541 aagctgcccg tgccctggcc caccctcgtg accaccttca cctacggcgt gcagtgcttc
      601 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      661 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      721 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      781 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      841 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      901 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      961 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1021 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac
     1081 ggcatggacg agctgtacaa gtaaagcggc cgcaaattgc ccagaattaa ttccccaact
     1141 gcacagaaca aatttccctt tgcttttgtg tagacctcaa aactttatcc cccatctttg
     1201 atcttatccc ttgtttttct gcttttttct tctttcttgg gttttgattt ccggacttaa
     1261 cgtttgtttt ccggtttgcg agacatattc tatcggattc tcaactgtct gatgaaataa
     1321 atatgtaatg ttctataagt ctttcaattt gatgggatgt tccacttctg tataggagcc
     1381 aattgattga caacatgcat caatcggagc tccagctttt gttcccttta gtgagggtta
     1441 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1501 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1561 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1621 tcgtgccagc tgcattaatg aatcggccaa cgctcgggga gaggcggttt gcgtattggg
     1681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1801 aagaacatga aggccttgac aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt
     1861 gtttgagatc tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2341 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2401 aagccagtta ccttcggaag aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2581 gggattttgg tcattagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2641 tgaagtttta aatcaatcta aagtatatat gtgtaacatt ggtctagtga ttatttgccg
     2701 actaccttgg tgatctcgcc tttcacgtag tgaacaaatt cttccaactg atctgcgcgc
     2761 gaggccaagc gatcttcttg tccaagataa gcctgcctag cttcaagtat gacgggctga
     2821 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
     2881 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
     2941 gcccagtcgg gcggcgagtt ccatagcgtt aaggttccat ttagcgcctc aaatagatcc
     3001 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
     3061 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     3121 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     3181 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     3241 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     3301 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     3361 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     3421 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata
     3481 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     3541 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat
     3601 cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc
     3661 gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac
     3721 cgtggctccc tcactttctg gctggatgat ggggcgattc aggcgatccc catccaacag
     3781 cccgccgtcg agcgggcttt tttatccccg gaagcctgtg gatagagggt agttatccac
     3841 gtgaaaccgc taatgccccg caaagccttg attcacgggg ctttccggcc cgctccaaaa
     3901 actatccacg tgaaatcgct aatcagggta cgtgaaatcg ctaatcggag tacgtgaaat
     3961 cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac gtgagaacgc taatagccct
     4021 ttcagatcaa cagcttgcaa acacccctcg ctccggcaag tagttacagc aagtagtatg
     4081 ttcaattagc ttttcaatta tgaatatata tatcaattat tggtcgccct tggcttgtgg
     4141 acaatgcgct acgcgcaccg gctccgcccg tggacaaccg caagcggttg cccaccgtcg
     4201 agcgcctttg cccacaaccc ggcggccggc cgcaacagat cgttttataa attttttttt
     4261 tttgaaaaag aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc
     4321 ggaattagat cttggcagga tatattgtgg tgtaacgtta tcagcttgca tgccggtcga
     4381 tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg
     4441 ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata
     4501 aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat
     4561 atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg
     4621 ttttaacgat ctgcttgact ctaggggtca tcagatttcg gtgacgggca ggaccggacg
     4681 gggcggcacc ggcaggctga agtccagctg ccagaaaccc acgtcatgcc agttcccgtg
     4741 cttgaagccg gccgcccgca gcatgccgcg gggggcatat ccgagcgcct cgtgcatgcg
     4801 cacgctcggg tcgttgggca gcccgatgac agcgaccacg ctcttgaagc cctgtgcctc
     4861 cagggacttc agcaggtggg tgtagagcgt ggagcccagt cccgtccgct ggtggcgggg
     4921 ggagacgtac acggtggact cggccgtcca gtcgtaggcg ttgcgtgcct tccagggacc
     4981 cgcgtaggcg atgccggcga cctcgccgtc cacctcggcg acgagccagg gatagcgctc
     5041 ccgcagacgg acgaggtcgt ccgtccactc ctgcggttcc tgcggctcgg tacggaagtt
     5101 gaccgtgctt gtctcgatgt agtggttgac gatggtgcag accgccggca tgtccgcctc
     5161 ggtggcacgg cggatgtcgg ccgggcgtcg ttctgggctc atggtagatc ccctcgatcg
     5221 agttgagagt gaatatgaga ctctaattgg ataccgaggg gaatttatgg aacgtcagtg
     5281 gagcattttt gacaagaaat atttgctagc ttatagtgac cttaggcgac ttttgaacgc
     5341 gcaataatgg tttctgacgt atgtgcttag ctcattaaac tccagaaacc cgcggctcag
     5401 tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca
     5461 tcggcggggg tcataacgtg actcccttaa ttctccgctc atgatcgata acattaacgc
     5521 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5581 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5641 aacgccaggg ttttcccagt cacgacgtt
//
