LOCUS       sequence-401115-        4527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Base plasmid for Plant STARR-seq terminator libraries..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4527)
  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 4527)
  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
FEATURES             Location/Qualifiers
     source          1..4527
                     /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(283..303)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(283..301)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(322..338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(322..338)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(335..357)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    346..362
                     /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(370..376,377..394,395..400))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    415..436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(553..570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    675..699
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(790..1378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(870..889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1549..2337)
                     /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      2628..3063
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    3191..3213
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(3237..3489)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(3509..4060)
                     /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(4102..4281)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     join(4513..4527,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4527,1..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 tgccgtcttc
      121 agtgcctgca ggcatagaag acccgtgacg agaccacaag gcgcgcctag tggtctccta
      181 agggagacgg gatcctacag ctagccgtct cgaggtgttc cacttctgta taggagccaa
      241 ttgattgaca acatgcatca atcggagctc cagcttttgt tccctttagt gagggttaat
      301 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      361 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      421 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      481 gtgccagctg cattaatgaa tcggccaacg ctcggggaga ggcggtttgc gtattgggcg
      541 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      661 gaacatgaag gccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
      721 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1201 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1261 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1321 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1381 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1441 gattttggtc attagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1501 aagttttaaa tcaatctaaa gtatatatgt gtaacattgg tctagtgatt atttgccgac
     1561 taccttggtg atctcgcctt tcacgtagtg aacaaattct tccaactgat ctgcgcgcga
     1621 ggccaagcga tcttcttgtc caagataagc ctgcctagct tcaagtatga cgggctgata
     1681 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     1741 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     1801 ccagtcgggc ggcgagttcc atagcgttaa ggttccattt agcgcctcaa atagatcctg
     1861 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     1921 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     1981 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     2041 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     2101 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     2161 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     2221 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     2281 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataac
     2341 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     2401 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     2461 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     2521 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     2581 tggctccctc actttctggc tggatgatgg gacgattcag gcgatcccca tccaacagcc
     2641 cgccgtcgag cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt
     2701 gaaaccgcta atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac
     2761 tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg
     2821 ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt
     2881 cagatcaaca gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt
     2941 caattagctt ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac
     3001 aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag
     3061 cgcctttgcc cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt
     3121 tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct tctggatttc cgatccccgg
     3181 aattagatct tggcaggata tattgtggtg taacgttatc agcttgcatg ccggtcgatc
     3241 tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt
     3301 ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa
     3361 taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca gaaattatat
     3421 gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt
     3481 ttaacgatct gcttgactct aggggtcatc agatttcggt gacgggcagg accggacggg
     3541 gcggcaccgg caggctgaag tccagctgcc agaaacccac gtcatgccag ttcccgtgct
     3601 tgaagccggc cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg tgcatgcgca
     3661 cgctcgggtc gttgggcagc ccgatgacag cgaccacgct cttgaagccc tgtgcctcca
     3721 gggacttcag caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg tggcgtggag
     3781 atacgtacac ggtggactcg gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg
     3841 cgtaggcgat gccggcgacc tcgccgtcca cctcggcgac gagccaggga tagcgctccc
     3901 gcagacggac gaggtcgtcc gtccactcct gcggttcctg cggctcggta cggaagttga
     3961 ccgtgcttgt ctcgatgtag tggttgacga tggtgcagac cgccggcatg tccgcctcgg
     4021 tggcacggcg gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc ctcgatcgag
     4081 ttgagagtga atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga
     4141 gcatttttga caagaaatat ttgctagctt atagtgacct taggcgactt ttgaacgcgc
     4201 aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg cggctcagtg
     4261 gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc
     4321 ggcgggggtc ataacgtgac tcccttaatt ctccgctcat gatcgataac attaacgctt
     4381 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4441 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     4501 cgccagggtt ttcccagtca cgacgtt
//
