LOCUS       sequence-274480-        6068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  high-copy URA-selectable plasmid, Mut ECM33 intron (unstable) 
            inserted into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6068)
  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 6068)
  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: pRS426-ECM_unstable
FEATURES             Location/Qualifiers
     source          1..6068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        157..372
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(1656..2111)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1793..1814)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2005..2024
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2237..2259
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2251..2268
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2252..2268
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2278..2296
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2298..2318)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2298..2316)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2337..2353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2337..2353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2350..2372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2361..2377
                     /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(2385..2391,2392..2409,2410..2415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2431..2451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2568..2585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2739..3327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2819..2838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3498..4289,4290..4358))
                     /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     4121..4140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4359..4463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5083..5960
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     5997..6015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6057..6068,1..11))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcagctcccg gagatctcgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
       61 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
      121 agagcagatt gtaccgagag tgcaccatac caccttttca attcatcatt ttttttttat
      181 tctttttttt gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
      241 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
      301 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa
      361 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
      421 gttggtatgt acacattctc cttttttttc atcttttttt ctttatttgc ctctttcctc
      481 acaaatctcg agtagattcg tggtcctctt cattcttttc ttttttcttt gtcgattact
      541 gggctttttt tcataggtct cgattgacgc ggacggacaa tgcgaaaaaa aaaatttcca
      601 aaagaggaaa cgggtttcga gatctcgtga ggatggtttt ggaatattct tcaatatttc
      661 tgcctgtcat accattaggg cgagtaatga aacagaataa tacatgtata aatcgatcgg
      721 gaatactaac actacttttc tttatctaac actacttttc tttatctaag cagctgccaa
      781 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac
      841 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac
      901 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt
      961 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac
     1021 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa
     1081 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt
     1141 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc
     1201 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat
     1261 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat
     1321 gacacccggt gtgggtttag atgacaaggg tgatgcattg ggtcaacagt atagaaccgt
     1381 ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa
     1441 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt
     1501 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa
     1561 ctcacaaatt agagcttcaa tttaattata tcagttatta ccctatgcgg tgtgaaatac
     1621 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     1681 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     1741 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     1801 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     1861 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     1921 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     1981 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2041 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2101 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2161 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2221 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
     2281 tacgactcac tataggccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     2341 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2401 agcataaagt gtaaagcctg gggtgcctag tgagtgagct aactcacatt aattgcgttg
     2461 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgc ttatcgatga
     4501 taagctgtca aacatgagaa ttaattccac ggactataga ctatacctag tatactccgt
     4561 ctactgtacg atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt
     4621 tgttactcta ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat
     4681 gtagtaaaac tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg
     4741 ctgccatcat tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt
     4801 gaggagatac agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac
     4861 ccatcattga attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt
     4921 gaacctgtat aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt
     4981 cctggagaaa ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat
     5041 tttctgcgtt tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt
     5101 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     5161 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     5221 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     5281 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     5341 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     5401 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     5461 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     5521 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     5581 tttttcgaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     5641 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     5701 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     5761 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     5821 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     5881 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     5941 gtggaagcgg tattcgcaat taagaaacca ttattatcat aacattaacc tataaaaata
     6001 ggcgtatcac gaggcccttt cgtcgaattc gcgcgtttcg gtgatgacgg tgaaaacctc
     6061 tgacacat
//
