LOCUS       sequence-334799-        6409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT U6 with a U57C mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6409)
  AUTHORS   van der Feltz C, Nikolai B, Schneider C, Paulson JC, Fu X, Hoskins 
            AA
  TITLE     Saccharomyces cerevisiae Ecm2 Modulates the Catalytic Steps of 
            pre-mRNA Splicing.
  JOURNAL   RNA. 2021 Feb 5. pii: rna.077727.120. doi: 10.1261/rna.077727.120.
  PUBMED    33547186
REFERENCE   2  (bases 1 to 6409)
  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: "RNA. 2021 
            Feb 5. pii: rna.077727.120. doi: 10.1261/rna.077727.120."
COMMENT     Sequence Label: U6-U57C
FEATURES             Location/Qualifiers
     source          1..6409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     182..198
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     183..199
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(517..533)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(517..533)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(565..584)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(566..584)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(591..608)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(591..607)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(600..622)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      748..1203
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(835..854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1045..1066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1306..1980)
                     /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"
     promoter        complement(1981..2261)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2282..2301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2401..2423
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2461..2479)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2520..3023
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4398..4502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4503..4571,4572..5363)
                     /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(4721..4740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5534..6122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6023..6042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6276..6293
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggtac cgggcccccc
      181 ctcgaggtcg acggtatcga taagcttgat atcgaattct tcggctacta taaataaatg
      241 tttttttcgc aactatgtgt tcgcgaagta acccttcgtg gacatttggt caatttgaaa
      301 caatacagag atgaccagca gttcccctgc ataaggatga accgttttac aaagagattt
      361 atttcgtttt tttttatcat cgagtgaagt atcgtgactt gtacatttga agatacccag
      421 cgtacagcag tgtatcttta tcttcctgta tgatatagat aactaacatc tcgaatagaa
      481 aattgtctcg cgttcgaacc taagcctttg ctatcggatc cactagttct agagcggccg
      541 ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
      601 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      661 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      721 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
      781 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      841 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      901 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      961 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1021 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1081 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1141 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1201 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagatcg
     1261 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     1321 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     1381 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     1441 tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa cccagtcaga
     1501 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
     1561 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
     1621 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
     1681 atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg
     1741 ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt
     1801 tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac
     1861 acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg cctctgtgct
     1921 ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat
     1981 actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg
     2041 ccctccctct tggccctctc cttttctttt ttcgaccgaa ttaattctta atcggcaaaa
     2101 aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa gctatttttc aataaagaat
     2161 atcttccact actgccatct ggcgtcataa ctgcaaagta cacatatatt acgatgctgt
     2221 tctattaaat gcttcctata ttatatatat agtaatgtcg tgatctatgg tgcactctca
     2281 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     2341 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2401 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     2461 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgg
     2521 atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat
     2581 ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa
     2641 ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa
     2701 aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata gatatacatt
     2761 cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga
     2821 caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta
     2881 tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt
     2941 taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa atttaaatta
     3001 taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc
     3061 gcggaaccct gatgaatccc ctaatgattt ttatcaaaat cattaagtta aggtagatac
     3121 acatcttgtc atatgatcaa atggtttcgc caaaaatcaa taatcagaca acaaaatgtg
     3181 cgaactcgat attttacacg actctcttta ccaattctgc cccgaattac acttaaaacg
     3241 actcaacagc ttaacgttgg cttgccacgc cttacttgac tgtaaaactc tcactcttac
     3301 cgaacttggc cgtaacctgc caaccaaagc gagaacaaaa cataacatca aacgaatcga
     3361 ccgattgtta ggtaatcgtc acctccacaa agagcgactc gctgtatacc gttggcatgc
     3421 tagctttatc tgttcgggca atacgatgcc cattgtactt gttgactggt ctgatatccg
     3481 tgagcaaaaa cggcttatgg tattgcgagc ttcagtcgca ctacacggtc gttctgttac
     3541 tctttatgag aaagcgttcc cgctttcaga gcaatgttca aagaaagctc atgaccaatt
     3601 tctagccgac cttgcgagca ttctaccgag taacaccaca ccgctcattg tcagtgatgc
     3661 tggctttaaa gtgccatggt ataaatccgt tgagaagctg ggttggtact ggttaagtcg
     3721 agtaagagga aaagtacaat atgcagacct aggagcggaa aactggaaac ctatcagcaa
     3781 cttacatgat atgtcatcta gtcactcaaa gactttaggc tataagaggc tgactaaaag
     3841 caatccaatc tcatgccaaa ttctattgta taaatctcgc tctaaaggcc gaaaaaatca
     3901 gcgctcgaca cggactcatt gtcaccaccc gtcacataaa atctactcag cgtcggcaaa
     3961 ggagccatgg attctagcaa ctaacttacc tgttgaaatt cgaacaccca aacaacttgt
     4021 taatatctat tcgaagcgaa tgcagattga agaaaccttc cgagacttga aaagtcctgc
     4081 ctacggacta ggcctacgcc atagccgaac gagcagctca gagcgttttg atatcatgct
     4141 gctaatcgcc ctgatgcttc aactaacatg ttggcttgcg ggcgttcatg ctcagaaaca
     4201 aggttgggac aagcacttcc aggctaacac agtcagaaat cgaaacgtac tctcaacagt
     4261 tcgcttaggc atggaagttt tgcggcattc tggctacaca ataacaaggg aagacttact
     4321 cgtggctgca accctactag ctcaaaattt attcacacat ggttacgctt tggggaaatt
     4381 atgaggggat ctctcagcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4441 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4501 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5041 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5161 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5221 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5341 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5401 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6061 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6121 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6181 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6241 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6301 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6361 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaat
//
