LOCUS       sequence-393231-        5857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pol II expression of U6 Lsm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5857)
  AUTHORS   Lipinski KA, Chi J, Chen X, Hoskins AA, Brow DA
  TITLE     Yeast U6 snRNA made by RNA polymerase II is less stable but 
            functional.
  JOURNAL   RNA. 2022 Dec;28(12):1606-1620. doi: 10.1261/rna.079328.122. Epub 
            2022 Oct 4.
  PUBMED    36195346
REFERENCE   2  (bases 1 to 5857)
  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: 
            "10.1261/rna.079328.122"; journalName: "RNA"; date: "2022-12"; 
            volume: "28"; issue: "12"; pages: "1606-1620"
COMMENT     Sequence Label: RS314-U6(PolII-Lsm)
FEATURES             Location/Qualifiers
     source          1..5857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     162..181
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(400..504)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    541..1044
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     1085..1103
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1141..1163)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1263..1282
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1303..1583
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1584..2258
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(2361..2816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2498..2519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2710..2729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2942..2964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2956..2973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2957..2973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2980..2999
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2980..2998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3031..3047
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3031..3047
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3081..3097)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3082..3098)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3255..3372
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        complement(4196..4214)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4235..4251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4235..4251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4248..4270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4259..4275
                     /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(4283..4289,4290..4307,4308..4313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4328..4349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4466..4483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4637..5225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4717..4736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5396..5857,1..330,331..399))
                     /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 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
       61 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      121 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
      181 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      241 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      301 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      361 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      421 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      481 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc
      541 atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa
      601 atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag
      661 actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat
      721 taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt
      781 tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata
      841 aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt
      901 cttcattccg taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat
      961 aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg
     1021 cgtgtaagtt acaggcaagc gatccgtcta agaaaccatt attatcatga cattaaccta
     1081 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1141 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1201 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1261 tgcggcatca gagcagattg tactgagagt gcaccataga tcacgacatt actatatata
     1321 taatatagga agcatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac
     1381 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
     1441 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
     1501 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
     1561 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
     1621 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
     1681 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
     1741 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
     1801 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
     1861 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
     1921 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
     1981 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
     2041 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
     2101 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     2161 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     2221 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     2281 ttaagtattg tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa
     2341 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2401 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2461 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2521 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2581 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2641 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2701 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2761 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     2821 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2881 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2941 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     3001 gaattggagc tccaccgcgg tggcggccgc tctagaacta gtggatcccc cgggctgcag
     3061 gaattcgata tcaagcttat cgataccgtc gacctcgaga tcatattaca tggcattacc
     3121 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
     3181 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
     3241 gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg acggaagact
     3301 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
     3361 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
     3421 aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta acaaccatag
     3481 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
     3541 atttttgatc tattaacaga tataaaagga acagaatatt agttagcttc attgtccaat
     3601 tactattgtt acttacgttt aatgcatttt cttttctttc acttccttcc ttaatactcc
     3661 gttcgcgaag taacccttcg tggacatttg gtcaatttga aacaatacag agatgatcag
     3721 cagttcccct gcataaggat gaaccgtttt acaaagagat ttatttcgtt ttaattctct
     3781 aatgtgagtt cacgttatag aagctggaat tgtaagtaat acagaaatgg caactaaaat
     3841 ggtaggcatg agtgttctcc ttcttgtgtg tctttctttt cacccgatac agtaaacttt
     3901 gtataggaga gttgaccaat tacctgacag tgccgtctat aaacgagacc tcaacttcac
     3961 ctcaaaaacc ataattttat ctattgataa ttatcattat ggtcattttc cactaaattc
     4021 atagttacct tactcttatg ctttgtattt gtaggtgtaa aaatgacgaa aaatttaatt
     4081 atctgaaaaa ttattgtgtg aaaaaacttg attttgctat aaccttatgt aaaaccatat
     4141 aaaagacaaa gacggtaaac taaaacagca gagaggaagg taccagcttt tgttcccttt
     4201 agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4261 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4321 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4381 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4441 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5041 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5281 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5341 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5461 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5521 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5581 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5641 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5701 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5761 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5821 gctccttcgg tcctccgatc gttgtcagaa gtaagtt
//
