LOCUS       sequence-401114-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fully assembled Plant STARR-seq plasmid with the 35S terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  AUTHORS   Gorjifard S, Jores T, Tonnies J, Mueth NA, Bubb K, Wrightsman T, 
            Buckler ES, Fields S, Cuperus JT, Queitsch C
  TITLE     Arabidopsis and maize terminator strength is determined by GC 
            content, polyadenylation motifs and cleavage probability.
  JOURNAL   Nat Commun. 2024 Jul 12;15(1):5868. doi: 10.1038/s41467-024-50174-7.
  PUBMED    38997252
REFERENCE   2  (bases 1 to 5667)
  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: "Nat 
            Commun."; date: "2024-07-12"; volume: "15(1):5868. doi"; pages: " 
            10.1038/s41467-024-50174-7"
COMMENT     Sequence Label: pPSt_35Spr-GFP-t35S
FEATURES             Location/Qualifiers
     source          1..5667
                     /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        291..337
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             join(431..433,434..1147)
                     /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(473..494)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(734..753)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1081..1102
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1423..1443)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1423..1441)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1462..1478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1462..1478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1475..1497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1486..1502
                     /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(1510..1516,1517..1534,1535..1540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1555..1576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1693..1710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1815..1839
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1930..2518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2010..2029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2689..3477)
                     /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      3768..4203
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4331..4353
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(4377..4629)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4649..5200)
                     /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(5242..5421)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     join(5653..5667,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5667,1..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 tgactcggaa
      121 aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa aggtggaccc ccacccacga
      181 ggagcatcgt ggaaaacgaa gtcgttccaa ccacggcttc aaagcaagtg gatggatgtg
      241 atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcctgt gcaagaccct
      301 tcctctatat aaggaagttc atttcatttg gagaggacac gcccgtccga actccgaacc
      361 ccagaacaga gcaaagcctc ctcggcctcc ctgtccccag ccttccccga tgaccataga
      421 cgtgatcgac gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      481 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      541 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      601 gcccaccctc gtgaccacct tcacctacgg cgtgcagtgc ttcagccgct accccgacca
      661 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      721 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      781 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      841 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
      901 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
      961 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1021 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1081 catggtcctg ctggagttcg tgaccgccgc cgggatcact cacggcatgg acgagctgta
     1141 caagtaagct ctagctagag tcgatcgaca agctcgagtt tctccataat aatgtgtgag
     1201 tagttcccag ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata
     1261 agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc
     1321 ctaaaaccaa aatccagtac taaaatccag ataggtgttc cacttctgta taggagccaa
     1381 ttgattgaca acatgcatca atcggagctc cagcttttgt tccctttagt gagggttaat
     1441 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1501 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1561 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1621 gtgccagctg cattaatgaa tcggccaacg ctcggggaga ggcggtttgc gtattgggcg
     1681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1801 gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     1861 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2401 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2581 gattttggtc attagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2641 aagttttaaa tcaatctaaa gtatatatgt gtaacattgg tctagtgatt atttgccgac
     2701 taccttggtg atctcgcctt tcacgtagtg aacaaattct tccaactgat ctgcgcgcga
     2761 ggccaagcga tcttcttgtc caagataagc ctgcctagct tcaagtatga cgggctgata
     2821 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     2881 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     2941 ccagtcgggc ggcgagttcc atagcgttaa ggttccattt agcgcctcaa atagatcctg
     3001 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     3061 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     3121 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     3181 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     3241 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     3301 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     3361 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     3421 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataac
     3481 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     3541 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     3601 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     3661 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     3721 tggctccctc actttctggc tggatgatgg gacgattcag gcgatcccca tccaacagcc
     3781 cgccgtcgag cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt
     3841 gaaaccgcta atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac
     3901 tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg
     3961 ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt
     4021 cagatcaaca gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt
     4081 caattagctt ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac
     4141 aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag
     4201 cgcctttgcc cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt
     4261 tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct tctggatttc cgatccccgg
     4321 aattagatct tggcaggata tattgtggtg taacgttatc agcttgcatg ccggtcgatc
     4381 tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt
     4441 ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa
     4501 taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca gaaattatat
     4561 gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt
     4621 ttaacgatct gcttgactct aggggtcatc agatttcggt gacgggcagg accggacggg
     4681 gcggcaccgg caggctgaag tccagctgcc agaaacccac gtcatgccag ttcccgtgct
     4741 tgaagccggc cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg tgcatgcgca
     4801 cgctcgggtc gttgggcagc ccgatgacag cgaccacgct cttgaagccc tgtgcctcca
     4861 gggacttcag caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg tggcgtggag
     4921 atacgtacac ggtggactcg gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg
     4981 cgtaggcgat gccggcgacc tcgccgtcca cctcggcgac gagccaggga tagcgctccc
     5041 gcagacggac gaggtcgtcc gtccactcct gcggttcctg cggctcggta cggaagttga
     5101 ccgtgcttgt ctcgatgtag tggttgacga tggtgcagac cgccggcatg tccgcctcgg
     5161 tggcacggcg gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc ctcgatcgag
     5221 ttgagagtga atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga
     5281 gcatttttga caagaaatat ttgctagctt atagtgacct taggcgactt ttgaacgcgc
     5341 aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg cggctcagtg
     5401 gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc
     5461 ggcgggggtc ataacgtgac tcccttaatt ctccgctcat gatcgataac attaacgctt
     5521 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5581 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     5641 cgccagggtt ttcccagtca cgacgtt
//
