LOCUS       sequence-273866-        4868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  low-copy URA-selectable plasmid, APS3 intron inserted into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4868)
  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 4868)
  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-APS3_intron
FEATURES             Location/Qualifiers
     source          1..4868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     502..520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(562..584)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     687..706
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        730..945
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(1957..2412)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2094..2115)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2306..2325
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2389..2667)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPNSPYSESYYARSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2538..2560
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2552..2569
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2553..2569
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2579..2598
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2579..2597
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2624..2644)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2624..2642)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2663..2679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2663..2679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2676..2698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2687..2703
                     /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(2711..2717,2718..2735,2736..2741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2756..2777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2894..2911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3065..3653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3145..3164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3824..4615,4616..4684))
                     /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     4447..4466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4685..4789)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(4826..4868,1..461)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
       61 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
      121 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
      181 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
      241 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
      301 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
      361 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
      421 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccattatt
      481 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtcg aattcgcgcg
      541 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagatct cggtcacagc
      601 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      661 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtaccga gagtgcacca
      721 taccaccttt tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga
      781 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
      841 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
      901 acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca
      961 tataaggaac gtgctgctac tcatcctagt cctgttggta tgtaagattt ctcaagggtc
     1021 ttttcttctg ccaaaattat acattactaa cataattcat catttttggt atagctgcca
     1081 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     1141 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     1201 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     1261 tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata
     1321 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     1381 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     1441 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     1501 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     1561 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     1621 tgacacccgg tgtgggttta gatgacaagg gtgatgcatt gggtcaacag tatagaaccg
     1681 tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     1741 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     1801 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     1861 actcacaaat tagagcttca atttaattat atcagttatt accctatgcg gtgtgaaata
     1921 ccgcacagat gcgtaagtag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     1981 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2041 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2101 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2161 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2221 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2281 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2341 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2401 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2461 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2521 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     2581 atacgactca ctatagggcg aattgggtac ccagcttttg ttccctttag tgagggttaa
     2641 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2701 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2761 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4801 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     4861 tttaatat
//
