LOCUS       sequence-245055-        3390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For run-off transcription of yeast RPL41A mRNA with and without an 
            82 nucleotide poly(A) tail..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3390)
  AUTHORS   Liu X, Schuessler PJ, Sahoo A, Walker SE
  TITLE     Reconstitution and analyses of RNA interactions with eukaryotic 
            translation initiation factors and ribosomal preinitiation 
            complexes.
  JOURNAL   Methods. 2019 Mar 26. pii: S1046-2023(18)30395-5. doi: 
            10.1016/j.ymeth.2019.03.024.
  PUBMED    30926531
REFERENCE   2  (bases 1 to 3390)
  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: "Methods. 
            2019 Mar 26. pii: S1046-2023(18)30395-5. doi: 
            10.1016/j.ymeth.2019.03.024."
COMMENT     Sequence Label: pSW104
FEATURES             Location/Qualifiers
     source          1..3390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(445..461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(445..461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(458..480)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    469..485
                     /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(493..499,500..517,518..523))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    538..559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(676..693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(847..1435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(927..946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1606..2397,2398..2466))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2229..2248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2467..2571)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2639..2657
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2695..2717)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2817..2836
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3050..3069
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3359..3377
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3359..3376
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taatacgact cactatagga gaccacatcg attcaatcga aatgagagcc aagtggagaa
       61 agaagagaac tagaagactt aagagaaaga gacggaaggt gagagccaga tccaaataag
      121 cggattatga gtaaataact ctaattttgt tttaaattct ttcaagagta tcgtaatgtc
      181 attgatgaat taacatgtta gtttctattc tacctcataa tggatctaaa ttgcatacta
      241 atctcacggt ggggtgtaaa ccattgccta ctatttatat agtgctttat atatgtctca
      301 catagtttaa tcaattgtcc gtttttttga aaaggatccc tgcagaaaaa aaaaaaaaaa
      361 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      421 aaaaaaaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2581 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2641 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     2701 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     2761 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     2821 atcagagcag attgtactga gagtgcacca ttcgacgctc tcccttatgc gactcctgca
     2881 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     2941 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     3001 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     3061 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     3121 cgtagaggat ctggctagcg atgaccctgc tgattggttc gctgaccatt tccgggtgcg
     3181 ggacggcgtt accagaaact cagaaggttc gtccaaccaa accgactctg acggcagttt
     3241 acgagagaga tgatagggtc tgcttcagta agccagatgc tacacaatta ggcttgtaca
     3301 tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat
     3361 ttaggtgaca ctatagaata cacggaattc
//
