LOCUS       sequence-273734-        5100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  low-copy URA-selectable plasmid, ACT1 intron inserted into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5100)
  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 5100)
  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-ACT1_intron
FEATURES             Location/Qualifiers
     source          1..5100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        139..354
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(1598..2053)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1735..1756)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1947..1966
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2030..2308)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPNSPYSESYYARSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2179..2201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2193..2210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2194..2210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2220..2239
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2220..2238
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2265..2285)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2265..2283)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2304..2320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2304..2320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2317..2339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2328..2344
                     /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(2352..2358,2359..2376,2377..2382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2397..2418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2535..2552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2706..3294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2786..2805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3465..4256,4257..4325))
                     /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     4088..4107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4326..4430)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4467..4970
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5011..5029
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5071..5093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
       61 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtaccgag
      121 agtgcaccat accacctttt caattcatca tttttttttt attctttttt ttgatttcgg
      181 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
      241 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
      301 gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg
      361 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttggtat gttctagcgc
      421 ttgcaccatc ccatttaact gtaagaagaa ttgcacggtc ccaattgctc gagagatttc
      481 tcttttacct ttttttacta tttttcactc tcccataacc tcctatattg actgatctgt
      541 aataaccacg atattattgg aataaatagg ggcttgaaat ttggaaaaaa aaaaaaaact
      601 gaaatatttt cgtgataagt gatagtgata ttcttctttt atttgctact gttactaagt
      661 ctcatgtact aacatcgatt gcttcattct ttttgttgct atattatatg tttagctgcc
      721 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
      781 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
      841 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
      901 ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat
      961 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     1021 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     1081 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     1141 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     1201 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     1261 atgacacccg gtgtgggttt agatgacaag ggtgatgcat tgggtcaaca gtatagaacc
     1321 gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg actatttgca
     1381 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     1441 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     1501 aactcacaaa ttagagcttc aatttaatta tatcagttat taccctatgc ggtgtgaaat
     1561 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     1621 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1681 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1741 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1801 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1861 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1921 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1981 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2041 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     2101 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     2161 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     2221 aatacgactc actatagggc gaattgggta cccagctttt gttcccttta gtgagggtta
     2281 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2341 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2401 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2461 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2521 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2881 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3121 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3361 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3421 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3721 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3901 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4201 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4321 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4441 cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat aatttaaatt
     4501 ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata
     4561 gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt
     4621 atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa
     4681 gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct
     4741 atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt
     4801 ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta
     4861 ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt
     4921 attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtctaagaa
     4981 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcga
     5041 attcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagatctc
//
