LOCUS       sequence-190770-        6484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human SF3B1 (AA 1-500) with N-term GST tag for inducible 
            expression in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6484)
  AUTHORS   Murthy T, Bluemn T, Gupta A, Reimer M, Rao S, Pillai MM, Minella AC
  TITLE     Cyclin-dependent kinase 1 (CDK1) and CDK2 have opposing roles in 
            regulating interactions of splicing factor 3B1 with chromatin.
  JOURNAL   J Biol Chem. 2018 May 15. pii: RA118.001654. doi: 
            10.1074/jbc.RA118.001654.
  PUBMED    29764937
REFERENCE   2  (bases 1 to 6484)
  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: "J Biol 
            Chem. 2018 May 15. pii: RA118.001654. doi: 
            10.1074/jbc.RA118.001654."
COMMENT     Sequence Label: pGEX-SF3B1-NTerm
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(52..57,58..75,76..82)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    90..106
                     /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     114..130
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     114..130
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             126..653
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(146..163)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(146..162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(155..177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(755..760,761..776,777..783)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    791..807
                     /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)."
     CDS             830..1483
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1441..1463
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1490..1513
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3106..3128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3273..3291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3359..3463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3464..3532,3533..4324)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3682..3701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4495..5083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4984..5003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5237..5254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5327..5404
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5405..6484,1..3)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5424..5443)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
       61 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      121 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      181 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      241 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      301 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
      361 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
      421 ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
      481 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
      541 agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg
      601 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
      661 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
      721 ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc ggctcgtata
      781 atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat
      841 actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct
      901 tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa
      961 aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa
     1021 attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg
     1081 ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata
     1141 cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag
     1201 caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa
     1261 tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata
     1321 catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga
     1381 agctatccca caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca
     1441 gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggaagttct
     1501 gttccagggg cccctgggat ccatggctaa gatcgctaaa acccacgagg acattgaggc
     1561 acagattcgg gagattcagg gcaagaaggc agcactggac gaggcacagg gcgtggggct
     1621 ggatagcacc ggctactatg accaggaaat ctacggcggg tctgacagta gattcgccgg
     1681 ctacgtgacc agcatcgccg ctaccgagct ggaagacgat gacgatgact atagctcctc
     1741 tacttcactg ctggggcaga agaaacctgg ataccacgca ccagtggccc tgctgaacga
     1801 tatcccacag tccaccgagc agtatgaccc cttcgctgaa caccggcccc ctaagattgc
     1861 agatagagag gacgaataca agaaacatcg gagaacaatg atcatttctc cagagcggct
     1921 ggaccccttc gccgacggag gcaaaacacc tgatccaaag atgaacgcca ggacttacat
     1981 ggacgtgatg cgcgaacagc atctgaccaa ggaggaacgg gagatcagac agcagctggc
     2041 cgagaaggca aaagccggag aactgaaagt cgtcaatggc gcagccgctt ctcagccacc
     2101 cagtaagagg aaaaggcgct gggatcagac cgctgaccag acacctggag caactccaaa
     2161 gaaactgagt tcatgggatc aggcagagac cccaggacac acacctagcc tgcgatggga
     2221 cgaaacccct ggacgagcaa agggctccga gactccagga gctaccccag gcagcaagat
     2281 ttgggacccc acaccttctc atactccagc tggcgcagca accccaggaa gaggcgacac
     2341 accaggacac gcaactccag gacatggagg agctacaagc tccgcaagaa aaaatcgctg
     2401 ggatgagacc cctaagacag aaagggacac tccagggcac ggatcaggct gggccgaaac
     2461 tccacgaacc gatcgaggag gggacagcat cggggagact ccaacccccg gagcttcaaa
     2521 gagaaaaagc agatgggatg agacacccgc atcccagatg ggaggctcta cccctgtgct
     2581 gacaccagga aagactccca tcggcacccc tgccatgaac atggctacac ctactccagg
     2641 ccatattatg tctatgaccc cagaacagct ccaggcctgg cgatgggagc gagaaatcga
     2701 cgagcgcaat cgacctctga gtgatgagga actggacgct atgttcccag agggctataa
     2761 ggtgctgcct ccacctgccg gctacgtccc aatcagaact cccgccagga aactgaccgc
     2821 tacacccact cctctggggg gaatgaccgg gtttcacatg cagacagagg atcgcactat
     2881 gaagagcgtg aacgaccagc cttccggaaa tctgcctttc ctgaagccag atgacatcca
     2941 gtactttgat aagctgctgg tggacgtgga cgaatccaca ctgtctccag aggaacagaa
     3001 ggagcgcaaa atcatgaagc tgtgagaatt cccgggtcga ctcgagcggc cgcatcgtga
     3061 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3121 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3181 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3241 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     3301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3421 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     4021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4621 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     5281 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     5341 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     5401 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     5461 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     5521 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5581 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     5641 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     5701 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     5761 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     5821 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     5881 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     5941 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     6001 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     6061 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     6121 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     6181 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     6241 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     6301 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     6361 gcccgtctca ctggtgaaaa gaaaaaccac gctagcgccc aatacgcaaa ccgcctctcc
     6421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6481 gcag
//
