LOCUS       sequence-273737-        4880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  low-copy URA-selectable plasmid, HNT2 intron inserted into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4880)
  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 4880)
  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-HNT2_intron
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(241..345)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    382..885
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     926..944
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(986..1008)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1111..1130
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1154..1369
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(2393..2848)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2530..2551)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2742..2761
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2825..3103)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPNSPYSESYYARSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2974..2996
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2988..3005
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2989..3005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3015..3034
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3015..3033
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3060..3080)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3060..3078)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3099..3115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3099..3115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3112..3134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3123..3139
                     /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(3147..3153,3154..3171,3172..3177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3192..3213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3330..3347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3501..4089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3581..3600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4260..4880,1..171,172..240))
                     /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"
ORIGIN
        1 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
       61 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
      121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
      181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
      241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
      301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      361 agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta
      421 atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt
      481 tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt
      541 gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca
      601 cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta
      661 atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta
      721 aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc
      781 taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc
      841 atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtct aagaaaccat
      901 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcgaattcg
      961 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag atctcggtca
     1021 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     1081 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ccgagagtgc
     1141 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
     1201 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     1261 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     1321 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
     1381 tacatataag gaacgtgctg ctactcatcc tagtcctgtt ggtatgcact ctcatatgtt
     1441 tttttgtacc ccattcgcac cattgcgtta ctaactatat ggagacctta taatctctgc
     1501 ttctttacag ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     1561 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     1621 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     1681 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt
     1741 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     1801 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     1861 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     1921 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     1981 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     2041 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggtga tgcattgggt
     2101 caacagtata gaaccgtgga tgatgtggtc tctacaggat ctgacattat tattgttgga
     2161 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     2221 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     2281 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     2341 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     2401 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2461 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2521 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2581 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2641 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2701 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2761 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2821 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     2881 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2941 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     3001 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtacccag cttttgttcc
     3061 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     3121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3241 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3301 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3361 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3421 ggggataacg caggaaagaa catgtgaaca aaaggccagc aaaaggccag gaaccgtaaa
     3481 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3541 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3601 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3661 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3721 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3781 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3841 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3901 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3961 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4021 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4081 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4141 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4201 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4261 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4321 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4381 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4441 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4501 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4561 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4621 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4681 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4741 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4801 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4861 tcttgcccgg cgtcaatacg
//
