LOCUS       sequence-348817-        7138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral construct to insert a N-terminal FLAG to one allele of 
            human SF3B1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  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 7138)
  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_FLAG_hSF3B1
FEATURES             Location/Qualifiers
     source          1..7138
                     /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
     CDS             1162..1185
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1186..1233
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     primer_bind     1320..1336
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(1338..1371)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          1409..1638
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1621..1640
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1767..2229
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1823..1840)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2050..2069
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2243..3037
                     /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(2297..2316)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2907..2926
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3059..3193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3083..3102)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3137..3156
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(3290..3323)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   4412..4541
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4603..5191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4683..4702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5362..6153,6154..6222))
                     /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     5985..6004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6223..6327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6395..6413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6451..6473)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6573..6592
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6609..7064)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6746..6767)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6958..6977
                     /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 ccggtcctcg gctgcttccg ggacccatct
      181 ttctgagcct ttctttcttt ttcatcgtcg aagtcaaaga ccagtgtgac agtacaaggt
      241 tatattataa ttgatgccca ggaaaacaga ctgccgtatt ctgatctatt aaaattcctg
      301 atttaggaat accgtctagg atttaagaga aacatctaat gaatgttaat aatgacgatg
      361 agtggttttc taggaaccca taaagtaaaa ggctttaaaa gggtggcaag attgtcactg
      421 tgcaactcag caacgatttg tgaggctcaa cgaaattcct acccaaatta tttgcaatta
      481 ttaaacagaa agaatgaaaa atgaccttga attacagggc gaaagatggg actgttaaga
      541 ctgtcttcgt aactaagcac tgtaaagact gagacagctg caagggaaag ttttcctcga
      601 gctcttccag tgaattttgg gaaaagggat tgaatgtctg aggtctcttc cagctctgaa
      661 accgcagcgt ttcgtacagt gttcattttt ttttaatggg aggggtctcg gggtcttgct
      721 ctgtcgcccg gtctggaggc ttgagtacag tggcgcgaca cgattacggc tcacggctgc
      781 ctcgaactcc tgggctcaag cgatgctccc gcctcagcct ccagagttgc tgggactcag
      841 tgtctgttct taactccctt cgtagaatcc actttcctcc agggcaaagt tactcactta
      901 aggaccttac ggctcggctc ggggatttcc tgggtcacca cgcaggcgac ttcagggccc
      961 aaccgtgtct cctcccccac ccacaatgca cctcgcggtt ccctctaggt ggcagttctg
     1021 tcacgtgtca cccgacggag cgcgctcccg tttggcgcac gcgcagagtg cagcccccag
     1081 ctatttttct ccgtggcggc ggcgacgagc ggaagttctt gggagcgcca gttccgtctg
     1141 tgtgttcgag tggacaaaat ggactacaag gacgatgacg acaagggttt gtaccgtaca
     1201 ccactgagac cgcggtggtt gaccagacaa accgcgaaga tcgccaagac tcacgaaggt
     1261 aagcggtctt tccctgctta cgtgttttct tcgttgctag cgtggggaga aggtctcact
     1321 ctagaactag tggatccata acttcgtata gcatacatta tacgaagtta tggaattaat
     1381 tcgctgtctg cgagggccag ctgttggggt gagtactccc tctcaaaagc gggcatgact
     1441 tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg
     1501 gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca
     1561 agcttgaggt gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact
     1621 ttgcctttct ctccacaggt gtccactccc aggtccaact gcaggtcgag cgtcgagaac
     1681 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     1741 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     1801 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     1861 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     1921 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     1981 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     2041 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     2101 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     2161 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     2221 cgatgataag cttgccacaa ccatggctga acaagatgga ttgcacgcag gttctccggc
     2281 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2341 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2401 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     2461 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2521 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2581 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2641 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     2701 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2761 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2821 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2881 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     2941 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3001 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcggggga
     3061 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3121 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3181 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3241 gtgggaggtt ttttctaggg ggccccggcg aggccgggat gagttgggaa taacttcgta
     3301 tagcatacat tatacgaagt tatccaaatt ttaaggtacc actgtgcata tgtatgaatt
     3361 cgatatcaag cttatcgata aaagcctttt ttcctaggat tctgcctttt tctatggttt
     3421 ctttcgtagg agctgagccc gcgtttcggg gccaagggtc gtctcctccg cgccagcggc
     3481 gcaggccact gccccgcgta atggcgtcct cctcttacac ccccgacagg cgctttgttc
     3541 agtgtttggg acctgctact tcagagtagc ttctcttctc tgaccagagg gcctcgttta
     3601 agcggcttcc tcagtaggat tcgtgggcaa gggatcgggg aagcgggcgt ggggagaagg
     3661 tctcactcgc ctctcgacac tgtcgtcccc atagtttgtg cagaagaggg tcagaaaagc
     3721 actgaaatac ggcagccgct tttccggcag actgaatcga atttggttaa accgagattc
     3781 cgttactcct acccttgtgt gtcttcaagc gtcagttact tgtttatttt tcttgcacga
     3841 aacgatttgg ggataaaatc tggtagttgt tttccctccc ttagtcactt ttagtataat
     3901 attgatcttt ttctttggag actggttatt ggaggagcag atggaagagg aaaacataaa
     3961 ttactttcat taatggccga agctttcaag ctagttgatc gttttatgga atacagagtg
     4021 tgcaaattag gaaaacgttg ggaatagact ggagttcatt aggaagtatc cagagatgac
     4081 agaaaatggg atcacttttc agaaaagacg aggcatgaga agttggaact gcattctttg
     4141 atttctgact tggaatccga gattttcact gagaacgaga cttctaagtc acttttgttt
     4201 catatgttct ttctgggtat ttggctcggt attagtctgt gagaggtaag agactgaggc
     4261 catccaaaat tttacagtct gataggagaa acagatgtgt atgttctcat tctctcatga
     4321 ggccagatgc atgcttaagt agaagaaaaa gcagcatgtt ctgggaacct gggtaacggg
     4381 tcgacctcga ggggccccgg ccggcggccg caggaacccc tagtgatgga gttggccact
     4441 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4501 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca
     4561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6361 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6421 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6481 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6541 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6601 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6661 cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata
     6721 gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6781 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6841 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     6901 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     6961 gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt
     7021 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     7081 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcc
//
