LOCUS       sequence-348816-        7133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral construct to change K to E at 700th amino acid (K700E) in
            human SF3B1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7133)
  AUTHORS   Boddu PC, Gupta AK, Kim JS, Neugebauer KM, Waldman T, Pillai MM
  TITLE     Generation of scalable cancer models by combining AAV-intron-trap, 
            CRISPR/Cas9, and inducible Cre-recombinase.
  JOURNAL   Commun Biol. 2021 Oct 13;4(1):1184. doi: 10.1038/s42003-021-02690-1.
  PUBMED    34645977
REFERENCE   2  (bases 1 to 7133)
  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: "Commun 
            Biol."; date: "2021-10-13"; volume: "4(1):1184. doi"; pages: " 
            10.1038/s42003-021-02690-1"
COMMENT     Sequence Label: pAAV_K700E_hSF3B1
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     primer_bind     1272..1288
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1272..1288
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(1290..1323)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          1361..1590
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1573..1592
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1719..2181
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1775..1792)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2002..2021
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2195..2989
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2249..2268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2859..2878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3011..3145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3035..3054)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3089..3108
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(3242..3275)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   4407..4536
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4598..5186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4678..4697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5357..6148,6149..6217))
                     /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     5980..5999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6218..6322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6390..6408
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6446..6468)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6568..6587
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6604..7059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6741..6762)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6953..6972
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca ccggtgcata aggtttgttc tccttaatag
      181 tgtttctgat agtttaaagt tttaactttt tcagttaatc aatgttaaat ttacatattc
      241 tcctagacct tttcctttgc acaagtacac accatacatg tacaaataca gttttataca
      301 gatgtacaga tgttagtttt tgtcatcttt gccgtaagag gttcatatta tttaattttt
      361 tcatttgctt ttattaaaat tagattagga tgagaacata tgagaactaa cgccagttag
      421 tattaagaac cagttttttc accgatggta attatggtgc agtatgttca tagggtgtat
      481 aagttatttc tttaaaaaat attgaatatt agtgtgtcac taatagctct cttaaatttt
      541 catctctgtc tggagtttga acttgtataa ttaaaattct taaacagttc gtcccttgat
      601 taacaaaagt cctgataatt ctaaatttct tatgtatgtt taattctgta catgagcatt
      661 tcatcagtaa ttgatgtgaa agtgtagctt cttctctttt ctctttttca gatcctcgtg
      721 gtcattgaac cgctattgat tgatgaagat tactatgcta gagtggaagg ccgagagatc
      781 atttctaatt tggcaaaggt atttacattt aatttctaga gaagaaaatt tatatctgtt
      841 tatggaattg attatggaaa gaaatggttg aagattaata ttaccaactc atgactgtcc
      901 tttctttgtt tacattttag gctgctggtc tggctactat gatctctacc atgagacctg
      961 atatagataa catggatgag tatgtccgta acacaacagc tagagctttt gctgttgtag
     1021 cctctgccct gggcattcct tctttattgc ccttcttaaa agctgtgtgc aaaagcaaga
     1081 agtcctggca agcgagacac actggtatta agattgtaca acagatagct attcttatgg
     1141 gctgtgccat cttgccacat cttagaagtt tagttgaaat cattgaacat ggtaagttgt
     1201 aatgtaactt tgtctttttt ttttttttct ttaggagaca gggtcttact atgttgccca
     1261 gactggagct ctctagaact agtggatcca taacttcgta tagcatacat tatacgaagt
     1321 tatggaatta attcgctgtc tgcgagggcc agctgttggg gtgagtactc cctctcaaaa
     1381 gcgggcatga cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc
     1441 acctggcccg cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga aaagacaatc
     1501 tttttgttgt caagcttgag gtgtggcagg cttgagatct ggccatacac ttgagtgaca
     1561 atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcaggtcg
     1621 agcgtcgaga acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1681 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1741 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1801 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1861 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1921 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     1981 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2041 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     2101 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2161 ctttgaaaaa cacgatgata agcttgccac aaccatggct gaacaagatg gattgcacgc
     2221 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     2281 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     2341 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     2401 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     2461 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     2521 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     2581 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     2641 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     2701 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     2761 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     2821 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     2881 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     2941 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     3001 gggttcgggg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3061 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3121 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3181 tcagggggag gtgtgggagg ttttttctag ggggccccgg cgaggccggg atgagttggg
     3241 aataacttcg tatagcatac attatacgaa gttatccaaa ttttaaggta ccactgtgca
     3301 tatgtatgaa ttcgatatca agcttatcga tggtcattga accgctattg gactcaaacc
     3361 tttgggctca agtgatcctc ctgctcagcc tccttagtag ttgggactag aggtacacac
     3421 acagcctgtc catgtttaat aggacagctg tcctaaaatt tgggctactg atttggggag
     3481 ataaatggaa aggcatagct ctacaaacta tagattttat gatgggtttg ttatattatc
     3541 tgctgacagg ctatggttca tgttttgctt ttacctaatt ttgtttaatg tgaacatatt
     3601 ctgcagtttg gctaaatagt tgatatattg agagaatctg gatgatattg tgtaacttag
     3661 gtaatgttgg ggcatagtta aaacctgtgt ttggttttgt aggtcttgtg gatgagcagc
     3721 aggaagttcg gaccatcagt gctttggcca ttgctgcctt ggctgaagca gcaactcctt
     3781 atggtatcga atcttttgat tctgtgttaa agcctttatg gaagggtatc cgccaacaca
     3841 gaggaaaggt aaatccacca attacctttt gatttatctt cattaaagtt aaggcgacat
     3901 aaatctaaat tactaaagta catatatttt ttatttaaaa atagggtttg gctgctttct
     3961 tgaaggctat tgggtatctt attcctctta tggatgcaga atatgccaac tactatacta
     4021 gagaagtgat gttaatcctt attcgagaat tccagtctcc tgatgaggaa atgaaaaaaa
     4081 ttgtgctgaa ggtaattatt ccagatttgt taatgtaaac tggatatgtt tcatggttct
     4141 aacagaattt taaagtgttt ttaaaaatca acaaaaaaaa ctgtatgtta cacgaatact
     4201 gtttgatttg tttttaaatt ttatttttta aaatcacgta atcagcaatg agtattctct
     4261 tcatttcagg tcagttgatt tattattact atttcttaac cttttaggtg gtaaaacagt
     4321 gttgtgggac agatggtcga cctcaatcac tagtgaattc gcggccgcct gcaggtcgac
     4381 ctcgaggggc cccggccggc ggccgcagga acccctagtg atggagttgg ccactccctc
     4441 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg
     4501 cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa
     4561 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4621 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4681 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4741 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4801 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4861 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4921 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4981 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5041 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5101 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5161 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5221 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5281 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5341 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5401 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5461 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5521 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5581 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5641 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5701 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5761 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5821 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5881 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5941 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6001 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6061 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6121 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6181 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6241 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6301 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6361 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     6421 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     6481 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6541 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6601 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6661 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
     6721 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6781 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6841 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6901 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6961 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     7021 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
     7081 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcc
//
