LOCUS       sequence-217641-        7010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6 snRNA coding sequence in pSE360(Ura) plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7010)
  AUTHORS   Madhani HD, Bordonne R, Guthrie C
  TITLE     Multiple roles for U6 snRNA in the splicing pathway.
  JOURNAL   Genes Dev. 1990 Dec;4(12B):2264-77.
  PUBMED    2149118
REFERENCE   2  (bases 1 to 7010)
  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"; journalName: "Genes 
            Dev."; date: "1990-12"; volume: "4(12B)"; pages: "2264-77"
COMMENT     Sequence Label: pHM4 (U6 in pSE360)
FEATURES             Location/Qualifiers
     source          1..7010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(810..828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(849..865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(849..865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(862..884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    873..889
                     /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(897..903,904..921,922..927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    942..963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1080..1097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1251..1839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1331..1350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2010..2801,2802..2870))
                     /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     2633..2652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2871..2975)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  2974..3741
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     misc_feature    3789..4292
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4333..4351
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4389..4411)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4511..4530
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4554..4769
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4770..5573
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5704..6159)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5841..5862)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6053..6072
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6285..6307
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6299..6316
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6300..6316
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6323..6342
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6323..6341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6374..6390
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6374..6390
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 actactgaat cgatgatact ttagagtttc attgcaacag tttcaacaca gcctggcatg
       61 aacagtggta aaagtatttc gtccactatt ttcggctact ataaataaat gtttttttcg
      121 catgcatgtg ttcgcgaagt aacccttcgt ggacatttgg tcaatttgaa acaatacaga
      181 gatgatcagc agttcccctg cataaggatg aaccgtttta caaagagatt tatttcgttt
      241 tttttttatc ctcgagtgaa gtatcgtgac ttgtacattt gaagataccc agcgtacagc
      301 agtgtatctt tatcttcctg tatgatatag ataactaaca tctcgaatag aaaattgtct
      361 cgcgttcgaa cctaagcctt tgctatcggt tgaacataga agaccgcgtg attcatttta
      421 gtgcgtaaga atgctattag tacttatgtg ctttatgaat gtgaatattg ttacagctat
      481 gcaatggtac gatcatactc tgaacgatcc atttaaatta tatatgtcaa acagaagaga
      541 aaggtttttt tattgaagat ttcacagaaa atcaatcatt ttgcaatatt ccttcgtccc
      601 agagaaggtt tcggctttta cagcaacgag aatgattctt ccattatttg tacctagctt
      661 ccacatggat ttcgataagt gtagtttcaa accatgaacg caattcaaaa ctttcaaatt
      721 ggtgcaatcc aagaccctcc ctttacattt ggagtcgtta tcaagcttat cgataccgtc
      781 gagggggggc ccggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
      841 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      901 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      961 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1021 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1321 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1381 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1441 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1501 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1561 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1621 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1681 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1741 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1801 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1861 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1921 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1981 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2041 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2101 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2161 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2221 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2281 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2341 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2401 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2461 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2521 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2581 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2641 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2701 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2761 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2821 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2881 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2941 tgtatttaga aaaataaaca aataggggtt ccgggtgatg ctgccaactt actgatttag
     3001 tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg
     3061 ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctatct
     3121 ctgctctcac tgccgtaaaa catggcaact gcagttcact tacaccgctt ctcaacccgg
     3181 tacgcaccag aaaatcattg atatggccat gaatggcgtt ggatgccggg caacagcccg
     3241 cattatgggc gttggcctca acacgatttt acgtcactta aaaaactcag gccgcagtcg
     3301 gtaacctcgc gcatacagcc gggcagtgac gtcatcgtct gcgcggaaat ggacgaacag
     3361 tggggctatg tcggggctaa atcgcgccag cgctggctgt tttacgcgta tgacagtctc
     3421 cggaagacgg ttgttgcgca cgtattcggt gaacgcacta tggcgacgct ggggcgtctt
     3481 atgagcctgc tgtcaccctt tgacgtggtg atatggatga cggatggctg gccgctgtat
     3541 gaatcccgcc tgaagggaaa gctgcacgta atcagcaagc gatatacgca gcgaattgag
     3601 cggcataacc tgaatctgag gcagcacctg gcacggctgg gacggaagtc gctgtcgttc
     3661 tcaaaatcgg tggagctgca tgacaaagtc atcgggcatt atctgaacat aaaacactat
     3721 caataagttg gagtcattac cggggttccg cgcacatttc cccgaaaagt gccacctggg
     3781 tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata
     3841 aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga
     3901 tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc
     3961 tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc
     4021 tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt
     4081 gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat
     4141 ttttttttct tcattccgta actcttctac cttctttatt tactttctaa aatccaaata
     4201 caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat
     4261 acgaggcgcg tgtaagttac aggcaagcga tccgtctaag aaaccattat tatcatgaca
     4321 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4381 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4441 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4501 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataccac cttttcaatt
     4561 catcattttt tttttattct tttttttgat ttcggtttcc ttgaaatttt tttgattcgg
     4621 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     4681 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     4741 caaaaacctg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     4801 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     4861 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     4921 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     4981 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag
     5041 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     5101 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     5161 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     5221 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     5281 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     5341 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     5401 acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta
     5461 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     5521 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     5581 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     5641 agttattacc ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     5701 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5761 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5821 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5881 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5941 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6001 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6061 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6121 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     6181 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6241 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     6301 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
     6361 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg atatcgtcca
     6421 ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt
     6481 tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct
     6541 cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg
     6601 gatcgataat tctccataat agttctgtta tttataatct ccagcactaa taaatgctat
     6661 acgtatattt gtacacaata taatttcaga atttatattg ctaccatgac tgtctgagaa
     6721 ttgggggaat aacttgataa ttgttgggat tccattgttc gtaaacgcaa taatattagg
     6781 tatatagaag atactaaatg ttctctccga ggatatagga atgctcacaa tggaatcgat
     6841 atatttctac ataatagtat tgagattatt cctcttttag ttttatataa ttcattatcc
     6901 tattacatta tcaatccttg catttcagct tccattagac ttaatgactg tttctcaatt
     6961 tttatgtcat cttcctggac ctcatgtgat actataccag tagcatgaat
//
