LOCUS       sequence-273868-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  low-copy URA-selectable plasmid, RPL27B intron inserted into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  AUTHORS   Morgan JT, Fink GR, Bartel DP
  TITLE     Excised linear introns regulate growth in yeast.
  JOURNAL   Nature. 2019 Jan;565(7741):606-611. doi: 10.1038/s41586-018-0828-1. 
            Epub 2019 Jan 16.
  PUBMED    30651636
REFERENCE   2  (bases 1 to 5175)
  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"; doi: 
            "10.1038/s41586-018-0828-1"; journalName: "Nature"; date: "2019-01";
            volume: "565"; issue: "7741"; pages: "606-611"
COMMENT     Sequence Label: pRS416-RPL27B_intron
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(14..118)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    155..658
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     699..717
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(759..781)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     884..903
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        927..1142
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(2461..2916)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2598..2619)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2810..2829
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2893..3171)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPNSPYSESYYARSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3042..3064
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3056..3073
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3057..3073
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3083..3102
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3083..3101
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3128..3148)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3128..3146)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3167..3183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3167..3183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3180..3202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3191..3207
                     /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(3215..3221,3222..3239,3240..3245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3260..3281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3398..3415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4328..5119,5120..5175,1..13))
                     /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     4951..4970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
       61 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
      121 catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa
      181 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
      241 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
      301 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
      361 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
      421 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
      481 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
      541 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
      601 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg
      661 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
      721 ttcgtcgaat tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      781 gagatctcgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      841 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      901 gtaccgagag tgcaccatac caccttttca attcatcatt ttttttttat tctttttttt
      961 gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     1021 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     1081 attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa
     1141 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttggtacgt
     1201 aaatttaaca gagcaaacgc tctaattaag atatcgaata aaaggggact ttcaggtgca
     1261 taagatggga aattgtacaa tctgagggac aaacaatatg gaaagcagta catgattgta
     1321 ttgtcattgt aggaggtacc aattgtgcca gtaaaaaaga gagggcccgg ggagcaccac
     1381 attcacgacc aatcgtgtcg atctgcaaaa aaggcacatg gaaaatgtaa tttatcagtg
     1441 ttcaacactg agttgaacag atagctatct aatcaatgtt aactttccag ggaacaaaaa
     1501 gctaaatcca cttctctttt atttcaaaat atcattagaa atagaaaaat ttactaacaa
     1561 attttcgtat tatcgtagct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     1621 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     1681 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     1741 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
     1801 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     1861 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     1921 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     1981 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     2041 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     2101 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggtgatg
     2161 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     2221 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     2281 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     2341 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     2401 tattacccta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2461 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2521 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2581 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2641 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2701 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2761 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2821 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2881 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
     2941 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3001 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3061 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtacccagct
     3121 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     3181 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3241 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3301 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3361 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgaacaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4321 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4381 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4441 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4501 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4561 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4621 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4681 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4741 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4801 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4861 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4921 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4981 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5041 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5101 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5161 gggcgacacg gaaat
//
