LOCUS       sequence-292043-        5654 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmid for STARR-seq in tobacco leaves. The GFP reporter gene is 
            under the control of a 35S minimal promoter and an upstream 35S core
            enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  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 5654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_35Senh(fwd)_35Spr+synJ
FEATURES             Location/Qualifiers
     source          1..5654
                     /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"
     promoter        274..320
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             join(366..368,369..1082)
                     /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(408..429)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(669..688)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1016..1037
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1403..1423)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1403..1421)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1442..1458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1442..1458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1455..1477)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1466..1482
                     /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(1490..1496,1497..1514,1515..1520))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1535..1556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1673..1690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1795..1819
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1910..2498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1990..2009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2669..3457)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3748..4183
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4318..4340
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(4364..4616)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4636..5187)
                     /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(5229..5408)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     join(5640..5654,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5654,1..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 tcagatctct
      121 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
      181 acgttccaac cacgtcttca aagcaagtgg attgatgtga catctccact gacgtaaggg
      241 atgacgcaca atcccactat ccttcaggag ctggcaagac ccttcctcta tataaggaag
      301 ttcatttcat ttggagagga cacgctggaa ttctagtata ctaaaccatg tattcttcct
      361 atttcgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      421 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      481 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      541 ccctcgtgac caccttcacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      601 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      661 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      721 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      781 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      841 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
      901 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      961 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1021 tcctgctgga gttcgtgacc gccgccggga tcactcacgg catggacgag ctgtacaagt
     1081 aaagcggccg caaattgccc agaattaatt ccccaactgc acagaacaaa tttccctttg
     1141 cttttgtgta gacctcaaaa ctttatcccc catctttgat cttatccctt gtttttctgc
     1201 ttttttcttc tttcttgggt tttgatttcc ggacttaacg tttgttttcc ggtttgcgag
     1261 acatattcta tcggattctc aactgtctga tgaaataaat atgtaatgtt ctataagtct
     1321 ttcaatttga tgggatgttc cacttctgta taggagccaa ttgattgaca acatgcatca
     1381 atcggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     1441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1621 tcggccaacg ctcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgaag gccttgacag
     1801 gatatattgg cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa
     1861 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1921 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1981 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2041 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2101 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2161 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2221 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2281 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2341 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa
     2401 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2461 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2521 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2581 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2641 gtatatatgt gtaacattgg tctagtgatt atttgccgac taccttggtg atctcgcctt
     2701 tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc
     2761 caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     2821 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     2881 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     2941 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     3001 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     3061 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     3121 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     3181 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     3241 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     3301 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     3361 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     3421 gttgagtcga tacttcggcg atcaccgctt ccctcataac accccttgta ttactgttta
     3481 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     3541 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     3601 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     3661 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     3721 tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag cgggcttttt
     3781 tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta atgccccgca
     3841 aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa
     3901 tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat
     3961 cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca gcttgcaaac
     4021 acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt ttcaattatg
     4081 aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc
     4141 tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgccagcgcc tttgcccaca
     4201 acccggcggc cggccgcaac agatcgtttt ataaattttt ttttttgaaa aagaaaaagc
     4261 ccgaaaggcg gcaacctctc gggcttctgg atttccgatc cccggaatta gagatcttgg
     4321 caggatatat tgtggtgtaa cgttatcagc ttgcatgccg gtcgatctag taacatagat
     4381 gacaccgcgc gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat
     4441 taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat
     4501 tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca
     4561 agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatctgct
     4621 tgactctagg ggtcatcaga tttcggtgac gggcaggacc ggacggggcg gcaccggcag
     4681 gctgaagtcc agctgccaga aacccacgtc atgccagttc ccgtgcttga agccggccgc
     4741 ccgcagcatg ccgcgggggg catatccgag cgcctcgtgc atgcgcacgc tcgggtcgtt
     4801 gggcagcccg atgacagcga ccacgctctt gaagccctgt gcctccaggg acttcagcag
     4861 gtgggtgtag agcgtggagc ccagtcccgt ccgctggtgg cggggggaga cgtacacggt
     4921 ggactcggcc gtccagtcgt aggcgttgcg tgccttccag ggacccgcgt aggcgatgcc
     4981 ggcgacctcg ccgtccacct cggcgacgag ccagggatag cgctcccgca gacggacgag
     5041 gtcgtccgtc cactcctgcg gttcctgcgg ctcggtacgg aagttgaccg tgcttgtctc
     5101 gatgtagtgg ttgacgatgg tgcagaccgc cggcatgtcc gcctcggtgg cacggcggat
     5161 gtcggccggg cgtcgttctg ggctcatggt agatcccctc gatcgagttg agagtgaata
     5221 tgagactcta attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa
     5281 gaaatatttg ctagcttata gtgaccttag gcgacttttg aacgcgcaat aatggtttct
     5341 gacgtatgtg cttagctcat taaactccag aaacccgcgg ctcagtggct ccttcaacgt
     5401 tgcggttctg tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata
     5461 acgtgactcc cttaattctc cgctcatgat cgataacatt aacgcttaca atttccattc
     5521 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5581 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5641 ccagtcacga cgtt
//
