LOCUS       sequence-217733-        8164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U2 snRNA with IIc+ mutation on a pRS315 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8164)
  TITLE     pRS315 U2 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8164)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pRS315 U2 IIc+
FEATURES             Location/Qualifiers
     source          1..8164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2115..2131)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2115..2131)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2163..2182)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2164..2182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2189..2206)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2189..2205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2198..2220)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2346..2801
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2643..2664
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3102..3509
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3510..4604
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(5098..5117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5217..5239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5277..5295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5337..5840
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5877..5981
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5982..6050,6051..6842)
                     /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     complement(6200..6219)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7013..7601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7502..7521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7755..7772
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7889..7910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7925..7930,7931..7948,7949..7955)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7963..7979
                     /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)."
     primer_bind     7968..7990
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7987..8003
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7987..8003
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8024..8042
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     8070..8086
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8071..8087
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtgctgcaca acaaggatgg gttcttggcc cggcaagaat ttctgcagcg ggtcgggtct
       61 gcagaggacg agagatacaa agaattgcgc cggcaacaac ttgctcagca gttgcagcag
      121 gatagcgaag cttcatagac aaatattaca tgtattaaat aacatgcttt tctgctcaaa
      181 tttttttaac cgccgaggcc tgcacacttt cactacgtgt ataacgatgc tgcagacaaa
      241 gtaatggggt gtatgggtgt gggtggcaaa aaaaacctag caacgctcta tgtttctttt
      301 cttttttctt tctttttttt tttttgccgt ttccgatggg ccactcggtg aaaacatata
      361 aaaagggctc tacattcatt ttttcatcga tgagtacttt acttgttatc agatttattc
      421 attttgtttc tacttgtttt ttttttaaat ccccacgaat ctctttgcct tttggcttag
      481 atcaagtgta gtatctgttc ttttcagtgt aacaactgaa atgacctcaa tgaggctcat
      541 taccttttca gttgttacaa tacacatttt ttggcaccca aaataataaa atggacggga
      601 agagactttt taagcaagtt gttttccgct aatgtcaggt ctcactactt tttgctgcta
      661 tttttcttcg ctcatggttt cttcataagg cgtttttatg atggtttttc gaaattggtt
      721 tttgagacga cggttgctca aggttattgt ttttgttttc ttctggttgt tttctatttt
      781 ctttttttta gctttctgtt tctcccttag tttggctttt tgcttcatac tcttccctgt
      841 ctttccgagc cgtttatgtc caacgcggga tttggttttt ctttatcgat gggaagaaat
      901 ggtgctatag taggttggga gataatattt atggtatggg gtgctagtgc ggatggggcg
      961 ctcttattgt tgatttcttc gctcgtcttc tttttctggt ggcgctgcaa gaggaagttt
     1021 ttcgactttg ttatgatttt tggtttgcaa ggaaaggtgt cttacgattc tttttttgat
     1081 gtaataggat aagcttgctt atcccccaag tatcggccaa agttgttgat tttccttttg
     1141 aagtgtcctc ggtttgaggg ggtgtagggt ggggttggtc tacaataaga gtgttccatt
     1201 gttaacgtgc tggcgtcttt tactatattt tttttcccag tttattttgt gcttattttc
     1261 tcattgagga gaaggagctc ttctcgcagg atataaatgg aggtttgcta aaggggagga
     1321 gatgtgtttg tgagaatact gctgagagag ttctggaaga gaaaaaaagg aggcaatgga
     1381 aggcgtttgc tgggaaaaga gaagagccat gactgcatct gttgtttcaa ggccagtttt
     1441 attaaccgcc tatgtcatag aggcgttttt tttggaggga tttgaagaat gccggcggca
     1501 tcaagaaacg gacttgatgg ttgacgcctg tttttaaagt tagagacgtc gcgaccctcg
     1561 cacttgtgga gtcgttcttg acttttactt tggtcgcttg atgtttctct cgtcttcccg
     1621 ttcgctcttt tattttttta tttttttttt cctttgactt cgcatgaatg agttcattca
     1681 tgaagcgaaa ggagagattc tatataccac gcaaccaaat atgtaagtat ataaaatata
     1741 tgtatgtgtg tgcactaatt ggcggcatac tctttcttga cccgttgtga tagggcctta
     1801 gctgcttccc attgcctgtc ccgagtccag cgactcaaga cgttggacgt gcgcacgtac
     1861 tcacacatgg ccgaattggc ccttacgaat gagctcttgg ctacccctct tgctgcactc
     1921 gccacacacc ataactggta taattcgttt ctcatgctag tctctttttt tttttttact
     1981 tgtttccttt tcctttgttg cttatccgct ttctcgttct ctcctctgtt cttgcctcaa
     2041 ctcctcaaat ctctgctgtc ccacttacgt tttatttttt caccgtcccc tttccctgcc
     2101 ctactacccc gggggatcca ctagttctag agcggccgcc accgcggtgg agctccaatt
     2161 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     2221 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2281 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2341 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2401 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttccttttt
     2461 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2521 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2581 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2641 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2701 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2761 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     2821 cttacgcatc tgtgcggtat ttcacaccgc atatcgaccg gtcgaggaga acttctagta
     2881 tatctacata cctaatatta ttgccttatt aaaaatggaa tcccaacaat tacatcaaaa
     2941 tccacattct cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg
     3001 ttatatttat aggataatta tactctattt ctcaacaagt aattggttgt ttggccgagc
     3061 ggtctaaggc gcctgattca agaaatatct tgaccgcagt taactgtggg aatactcagg
     3121 tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt atttttttcc tcaacataac
     3181 gagaacacac aggggcgcta tcgcacagaa tcaaattcga tgactggaaa ttttttgtta
     3241 atttcagagg tcgcctgacg catatacctt tttcaactga aaaattggga gaaaaaggaa
     3301 aggtgagagc gccggaaccg gcttttcata tagaatagag aagcgttcat gactaaatgc
     3361 ttgcatcaca atacttgaag ttgacaatat tatttaagga cctattgttt tttccaatag
     3421 gtggttagca atcgtcttac tttctaactt ttcttacctt ttacatttca gcaatatata
     3481 tatatatatt tcaaggatat accattctaa tgtctgcccc taagaagatc gtcgttttgc
     3541 caggtgacca cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt
     3601 ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta
     3661 tcgatgctac aggtgttcca cttccagatg aggcgctgga agcctccaag aaggctgatg
     3721 ccgttttgtt aggtgctgtg ggtggtccta aatggggtac cggtagtgtt agacctgaac
     3781 aaggtttact aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact
     3841 ttgcatccga ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg
     3901 acttcgttgt tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaagacg
     3961 atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca
     4021 caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg
     4081 ataaagctaa tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca
     4141 agaacgaatt ccctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc
     4201 tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata
     4261 tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct
     4321 tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg
     4381 ctccagattt gccaaagaat aaggtcaacc ctatcgccac tatcttgtct gctgcaatga
     4441 tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa
     4501 aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag
     4561 tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt
     4621 tatgatattt gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac
     4681 aaaatggaat atgttcatag ggtagacgaa actatatacg caatctacat acatttatca
     4741 agaaggagaa aaaggaggat gtaaaggaat acaggtaagc aaattgatac taatggctca
     4801 acgtgataag gaaaaagaat tgcactttaa cattaatatt gacaaggagg agggcaccac
     4861 acaaaaagtt aggtgtaaca gaaaatcatg aaactatgat tcctaattta tatattggag
     4921 gattttctct aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac ctgaccattt
     4981 gatggagttt aagtcaatac cttcttgaac catttcccat aatggtgaaa gttccctcaa
     5041 gaattttact ctgtcagaaa cggccttaac gacgtagtcg atatggtgca ctctcagtac
     5101 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5161 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5221 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5281 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt aggacggatc
     5341 gcttgcctgt aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta
     5401 ctctgtgttt atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt
     5461 agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa
     5521 gcgtacttta catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga
     5581 ttaacgataa gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa
     5641 gatgaaacaa ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt
     5701 gttggcgatc cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa
     5761 tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa
     5821 ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg
     5881 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5941 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6001 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6061 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6121 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6181 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6241 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6301 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6361 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6421 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6481 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6541 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6601 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6661 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6721 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6781 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6841 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6901 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6961 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7021 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7081 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7141 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7201 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7261 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7321 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7381 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7441 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7501 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7561 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7621 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7681 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7741 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7801 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7861 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7921 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7981 ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa
     8041 gggaacaaaa gctggtaccg ggccccccct cgaggtcgac ggtatcgata agcttgatat
     8101 cgaattcctg cagcccctgg actgggctag ttatgtagac cgtgctggac ttaacgacga
     8161 attg
//
