LOCUS       sequence-210612-        8208 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 8208)
  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 8208)
  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..8208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /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(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1378..1399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1590..1609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1826..1848
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1840..1857
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1867..1886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5096..5161
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5434..5454)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5434..5452)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5473..5489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5473..5489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5486..5508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5497..5513
                     /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(5521..5527,5528..5545,5546..5551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5704..5721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5875..6463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5955..5974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6634..7425,7426..7494))
                     /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     7257..7276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7495..7599)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7636..8139
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8181..8199
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgagcatgta
     1921 ctttccgcct aactaccatc acctgtcttt atttattccg ctgacagata tgcacaacca
     1981 attatcgtct cttgaaagct ttttagacgt gattgctgtt atttggcacg ttgttagtat
     2041 ttaagcttat tcgggtttgc attgatattt gcccgagcag aaatctcctt ctcgatgagt
     2101 tagaactgaa acatctcttc aattacttat acataataaa catagaaagc caggacctgt
     2161 catcatgagt catccgattc aatttgtaaa cgctaataat agcgataaga gccaccaatt
     2221 aggcggtcaa tattctattc ctcaggattt gagggaaaac ctgcagaaag aagctgcacg
     2281 tattggtgag aacgaaaaag atgtgctaca ggaaaaaatg gaaactagga cagtgcaaaa
     2341 tcgagaggat tcataccaca aaagaaggtt tgatatgaag tttgagccgg attccgatac
     2401 acaaactgtt acgtcttcag aaaataccca agacgctgtc gtgccaagga aaagaaaatc
     2461 tcgttgggat gtaaaagggt acgagccacc tgatgaatct tcaacagcgg tcaaagaaaa
     2521 ctctgatagt gctttggtta atgtagaagg aatccatgat ttgatgtttt ttaaaccttc
     2581 tgaccataaa tatttcgccg acgttatatc caaaaaaccc atagatgagt taaataagga
     2641 tgaaaagaaa gaaagaaccc tgtcgatgtt acttctgaaa ataaaaaatg gcaatacagc
     2701 gagtaggagg acttctatgc ggattttaac cgataaagct gttacatttg gtccagaaat
     2761 gatatttaac cgcttattgc ccatattact agacagaagt ttggaagacc aggaaagaca
     2821 tctaatgatc aagaccatag atcgcgtcct ttaccagcta ggagatttaa caaaacctta
     2881 cgttcacaaa atacttgtag tagctgcccc attgctaata gatgaagatc caatggttcg
     2941 ttcaacaggc caggagataa tcaccaatct gtctacagtc gctggcctaa aaaccattct
     3001 aacagtaatg agacctgata ttgagaatga ggatgagtat gttagaaatg taacttcaag
     3061 agcggcggct gtggtggcaa aagcccttgg tgttaatcaa ttgcttcctt tcatcaatgc
     3121 agcatgcaaa agtaaaaaaa gttggcaggc ccgtcacacg ggcataaaga tcgtgcaaca
     3181 gatagccatc ttgatgggat gcgcgattct tccacacctg aggtccttag tcgagatcat
     3241 tgaacatgga ctggtggatg agcaacagaa agtacgtacg attagtgcgc tggctatcgc
     3301 cgcccttgct gaagcagcta ccccttatgg aattgagagc tttgatagtg tcctgaaacc
     3361 cctgtggaaa ggtattagac agcacagggg aaaaggactg gctgccttcc ttaaagcaat
     3421 aggatatctg ataccgctaa tggacgccga atacgctaac tattacacga gggaagttat
     3481 gctaatcctt atccgtgagt ttcagagtcc ggacgaagag atgaaaaaga tagtattgaa
     3541 ggttgtgaaa cagtgctgtg gcaccgatgg cgtggaggct aattatatca agacagagat
     3601 tctaccacca ttctttaagc atttttggca acatagaatg gcattggaca ggagaaacta
     3661 cagacaacta gttgacacta cggtcgagtt agctaataag gtgggtgctg ccgaaattat
     3721 tagccgtata gttgatgacc ttaaggatga ggctgaacaa taccgtaaaa tggttatgga
     3781 aaccattgaa aagattatgg gtaatctagg tgcggctgac atcgaccata aactagagga
     3841 acagttaatt gacggtatcc tgtacgcttt ccaagagcag actacagagg attccgtgat
     3901 gctgaacggt tttgggactg tcgtaaatgc attgggtaaa cgtgtaaaac cgtacttacc
     3961 tcaaatttgt ggcacggttt tatggagatt aaataacaaa agtgcaaagg ttcgtcaaca
     4021 agcagcagat ctaatctcca gaactgccgt tgtcatgaag acctgtcagg aagagaagct
     4081 tatgggccat ttgggtgtcg tcttgtatga atatttaggc gaagaatatc cagaggtatt
     4141 aggttctata ttaggcgctc ttaaagccat tgtcaacgta ataggtatgc acaaaatgac
     4201 accacctatc aaggaccttt taccaaggct gacaccaatc ttaaagaaca gacatgaaaa
     4261 ggtacaagaa aactgtatcg acttggtcgg aaggatagct gatagaggcg ccgaatatgt
     4321 gtcagccagg gagtggatga ggatttgctt cgaattactg gagcttctta aagcgcacaa
     4381 gaaggctatc agaagggcga cggtaaatac ttttgggtac attgcaaaag ccataggccc
     4441 gcatgacgtc ctggcgactc tactaaacaa tttgaaggta caggaaagac agaatagagt
     4501 ttgtactact gtagcaatag ccatcgttgc cgagacgtgt agcccgttca cggtgttacc
     4561 cgccctaatg aacgagtata gggtcccgga attaaatgtc caaaatggtg tacttaaatc
     4621 actatccttt ctgtttgaat acataggcga aatggggaaa gattacatct acgcagtaac
     4681 tccattactg gaggatgccc ttatggaccg tgatttggtc catagacaga ctgcttcagc
     4741 cgtggtgcag catatgtcct taggggtgta cggttttggt tgtgaagata gtctaaacca
     4801 ccttctgaac tacgtttggc ctaacgtctt cgagacgagc ccgcatgtta tccaagcagt
     4861 aatgggggcg ttggagggct tgagagtggc gataggcccc tgcagaatgc tgcagtattg
     4921 tttgcaagga ttgttccatc ctgctcgtaa agtcagagat gtgtattgga aaatctataa
     4981 tagcatatat ataggatcac aagatgccat ggtacctttt taccccgtta caccagacaa
     5041 caatgaagaa tatatagaag aactggattt agttctgggt ggttctggtg gttctgatta
     5101 caaagaccac gatggtgact acaaagacca tgatatagat tacaaggatg atgatgataa
     5161 atgaatatat atagagcata taaataacca caacttgtaa gaatatttta cttgacacac
     5221 ctatatcatc acgtcacgat gccttcatag attatgatga aatataaaca gaacctgcta
     5281 ttggaaaaaa aagggcagga aaaaattttg tgaaagtgag taaaagaaag atgaagagcg
     5341 agacatggat aataagaaat agatgtcact aatggacaat tggaagactg atatggaaag
     5401 ttacgatgaa ggaggagctc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     5461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5521 caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact
     5581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7621 acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
     7681 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
     7741 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
     7801 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
     7861 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
     7921 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
     7981 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
     8041 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
     8101 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa accattatta
     8161 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
