LOCUS       sequence-213671-        8211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human HEATs 5-20 have been inserted into Hsh155 3xFLAG..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8211)
  AUTHORS   Carrocci TJ, Paulson JC, Hoskins AA
  TITLE     Functional Analysis of Hsh155/SF3b1 Interactions with the U2 
            snRNA/Branch Site Duplex.
  JOURNAL   RNA. 2018 May 11. pii: rna.065664.118. doi: 10.1261/rna.065664.118.
  PUBMED    29752352
REFERENCE   2  (bases 1 to 8211)
  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. 2018 
            May 11. pii: rna.065664.118. doi: 10.1261/rna.065664.118."
COMMENT     Sequence Label: Hsh155 Hs 5-20 3xFLAG
FEATURES             Location/Qualifiers
     source          1..8211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2777..2842
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3114..3134)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3114..3132)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3153..3169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3153..3169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3166..3188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3177..3193
                     /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(3201..3207,3208..3225,3226..3231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3246..3267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3384..3401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3555..4143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3635..3654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4314..5105,5106..5174))
                     /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     4937..4956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5175..5279)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5316..5819
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5860..5878
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5916..5938)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6038..6057
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6078..6358
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6359..7033
                     /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(7136..7591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7273..7294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7485..7504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7717..7739
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7731..7748
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7732..7748
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7758..7777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7758..7776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaaactagg acagtgcaaa atcgagagga ttcataccac aaaagaaggt ttgatatgaa
       61 gtttgagccg gattccgata cacaaactgt tacgtcttca gaaaataccc aagacgctgt
      121 cgtgccaagg aaaagaaaat ctcgttggga tgtaaaaggg tacgagccac ctgatgaatc
      181 ttcaacagcg gtcaaagaaa actctgatag tgctttggtt aatgtagaag gaatccatga
      241 tttgatgttt tttaaacctt ctgaccataa atatttcgcc gacgttatat ccaaaaaacc
      301 catagatgag ttaaataagg atgaaaagaa agaaagaacc ctgtcgatgt tacttctgaa
      361 aataaaaaat ggcaatacag cgagtaggag gacttctatg cggattttaa ccgataaagc
      421 tgttacattt ggtccagaaa tgatatttaa ccgcttattg cccatattac tagacagaag
      481 tttggaagac caggaaagac atctaatgat caagaccata gatcgcgtcc tttaccagct
      541 aggagattta acaaaacctt acgttcacaa aatacttgta gtagctgccc cattgctaat
      601 agatgaagat ccaatggttc gttcaacagg ccaggagata atcaccaatc tgtctacagt
      661 cgctggccta aaaaccattc taacagtaat gagacctgat attgagaatg aggatgagta
      721 tgttagaaat gtaacttcaa gagcggcggc tgtggtggca aaagcccttg gtgttaatca
      781 attgcttcct ttcatcaatg cagcatgcaa aagtaaaaaa agttggcagg cccgtcacac
      841 gggcataaag atcgtgcaac agatagccat cttgatggga tgcgcgattc ttccacacct
      901 gaggtcctta gtcgagatca ttgaacatgg actggtggat gagcaacaga aagtacgtac
      961 gattagtgcg ctggctatcg ccgcccttgc tgaagcagct accccttatg gaattgagag
     1021 ctttgatagt gtcctgaaac ccctgtggaa aggtattaga cagcacaggg gaaaaggact
     1081 ggctgccttc cttaaagcaa taggatatct gataccgcta atggacgccg aatacgctaa
     1141 ctattacacg agggaagtta tgctaatcct tatccgtgag tttcagagtc cggacgaaga
     1201 gatgaaaaag atagtattga aggttgtgaa acagtgctgt ggcaccgatg gcgtggaggc
     1261 taattatatc aagacagaga ttctaccacc attctttaag catttttggc aacatagaat
     1321 ggcattggac aggagaaact acagacaact agttgacact acggtcgagt tagctaataa
     1381 ggtgggtgct gccgaaatta ttagccgtat agttgatgac cttaaggatg aggctgaaca
     1441 ataccgtaaa atggttatgg aaaccattga aaagattatg ggtaatctag gtgcggctga
     1501 catcgaccat aaactagagg aacagttaat tgacggtatc ctgtacgctt tccaagagca
     1561 gactacagag gattccgtga tgctgaacgg ttttgggact gtcgtaaatg cattgggtaa
     1621 acgtgtaaaa ccgtacttac ctcaaatttg tggcacggtt ttatggagat taaataacaa
     1681 aagtgcaaag gttcgtcaac aagcagcaga tctaatctcc agaactgccg ttgtcatgaa
     1741 gacctgtcag gaagagaagc ttatgggcca tttgggtgtc gtcttgtatg aatatttagg
     1801 cgaagaatat ccagaggtat taggttctat attaggcgct cttaaagcca ttgtcaacgt
     1861 aataggtatg cacaaaatga caccacctat caaggacctt ttaccaaggc tgacaccaat
     1921 cttaaagaac agacatgaaa aggtacaaga aaactgtatc gacttggtcg gaaggatagc
     1981 tgatagaggc gccgaatatg tgtcagccag ggagtggatg aggatttgct tcgaattact
     2041 ggagcttctt aaagcgcaca agaaggctat cagaagggcg acggtaaata cttttgggta
     2101 cattgcaaaa gccataggcc cgcatgacgt cctggcgact ctactaaaca atttgaaggt
     2161 acaggaaaga cagaatagag tttgtactac tgtagcaata gccatcgttg ccgagacgtg
     2221 tagcccgttc acggtgttac ccgccctaat gaacgagtat agggtcccgg aattaaatgt
     2281 ccaaaatggt gtacttaaat cactatcctt tctgtttgaa tacataggcg aaatggggaa
     2341 agattacatc tacgcagtaa ctccattact ggaggatgcc cttatggacc gtgatttggt
     2401 ccatagacag actgcttcag ccgtggtgca gcatatgtcc ttaggggtgt acggttttgg
     2461 ttgtgaagat agtctaaacc accttctgaa ctacgtttgg cctaacgtct tcgagacgag
     2521 cccgcatgtt atccaagcag taatgggggc gttggagggc ttgagagtgg cgataggccc
     2581 ctgcagaatg ctgcagtatt gtttgcaagg attgttccat cctgctcgta aagtcagaga
     2641 tgtgtattgg aaaatctata atagcatata tataggatca caagatgcca tggtaccttt
     2701 ttaccccgtt acaccagaca acaatgaaga atatatagaa gaactggatt tagttctggg
     2761 tggttctggt ggttctgatt acaaagacca cgatggtgac tacaaagacc atgatataga
     2821 ttacaaggat gatgatgata aatgaatata tatagagcat ataaataacc acaacttgta
     2881 agaatatttt acttgacaca cctatatcat cacgtcacga tgccttcata gattatgatg
     2941 aaatataaac agaacctgct attggaaaaa aaagggcagg aaaaaatttt gtgaaagtga
     3001 gtaaaagaaa gatgaagagc gagacatgga taataagaaa tagatgtcac taatggacaa
     3061 ttggaagact gatatggaaa gttacgatga aggaggagct ccagcttttg ttccctttag
     3121 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3181 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3241 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3301 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3361 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3421 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3481 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3541 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3601 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3721 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3961 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4321 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4381 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4441 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4501 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4561 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4621 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4681 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4741 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4801 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4861 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4921 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4981 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5041 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5101 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5161 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5221 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5281 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata
     5341 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
     5401 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
     5461 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
     5521 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
     5581 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
     5641 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     5701 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     5761 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     5821 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5881 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5941 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6001 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6061 agagtgcacc atagatcacg acattactat atatataata taggaagcat ttaatagaac
     6121 agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag tggaagatat
     6181 tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc ttttcttttt
     6241 ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca
     6301 ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat
     6361 gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag
     6421 cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt
     6481 gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa
     6541 agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc
     6601 taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg
     6661 agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt
     6721 atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat
     6781 tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc
     6841 tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact
     6901 gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag
     6961 cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa
     7021 tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgccg
     7081 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     7141 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7201 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7261 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7321 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7381 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7441 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7501 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7561 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     7621 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     7681 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     7741 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     7801 ctcgagcatg tactttccgc ctaactacca tcacctgtct ttatttattc cgctgacaga
     7861 tatgcacaac caattatcgt ctcttgaaag ctttttagac gtgattgctg ttatttggca
     7921 cgttgttagt atttaagctt attcgggttt gcattgatat ttgcccgagc agaaatctcc
     7981 ttctcgatga gttagaactg aaacatctct tcaattactt atacataata aacatagaaa
     8041 gccaggacct gtcatcatga gtcatccgat tcaatttgta aacgctaata atagcgataa
     8101 gagccaccaa ttaggcggtc aatattctat tcctcaggat ttgagggaaa acctgcagaa
     8161 agaagctgca cgtattggtg agaacgaaaa agatgtgcta caggaaaaaa t
//
