LOCUS       sequence-348819-        7011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral construct to change S to F at 34th amino acid (S34F) in 
            human U2AF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7011)
  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 7011)
  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_S34F_U2AF1
FEATURES             Location/Qualifiers
     source          1..7011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   225..354
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    1427..1460
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1557..1691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1594..1613)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1648..1667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1713..2507)
                     /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(1824..1843)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2434..2453
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_feature    2521..2983
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2681..2700)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2910..2927
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3110..3129)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3112..3341
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     protein_bind    3379..3412
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3414..3430)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3414..3430)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     repeat_region   4498..4638
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4498..4627
                     /label=AAV2 ITR
     rep_origin      4713..5168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4800..4819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5010..5031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5185..5204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5304..5326
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5364..5382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5450..5554
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5555..5623,5624..6415)
                     /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     complement(5773..5792)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6586..7011,1..163)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
       61 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
      121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
      241 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      301 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
      361 gccggccggg gcccctcgag gtcgacaatc catggccact ggtttagtta ggggagattc
      421 ggttcataaa tcaaaacatt aaagaaaaaa caattgcaat acacagaaaa tgtaaagcta
      481 gttctccaag attttgaaat ttgtaaaaag aaactcatgt tcagagctat caaaagacag
      541 actgctatta tactaaccac ttggtaatta gcataatgga gggaaaagcc atcatcctct
      601 tcccttctaa aaacactgtg tgctgtgggg tcgtccatgg gcttcaaccg gaaaaaaccc
      661 atgtttcctt ctcccagctc acttacacaa agtggtagat tttcttaaat gcttacagga
      721 aaacgtatac agacagatgt gagctcaaag gcagagcctg gcagctccca gggaccacgc
      781 ataatagcta ctcaacagct cagaggctgc acatttagac ttcaagggtg ggttggaagg
      841 agttccccag ccctccatct ggaaagactt gggcatcgat tttcaaattt caaatgttct
      901 catcaacgtg ggcacaagtt ttctgctccg attagaggtg ccttctccct ttaatccgta
      961 agtgatcctt tatagtgtac caacctttgc cttttcttca actaaccaaa aataaaacta
     1021 tcaaaactca caacttttga ttgaactgaa ctcagagcgg atctaagccc aagataacag
     1081 gcactcctca attaaccaaa cagacccgct aagagcaatt tacggcccat tttctacctt
     1141 tcagcttaat ttttcatgtt tccatgacgt gatgcgactc gctgtttcct cagctaggga
     1201 tgtgaaagtg aggacgcagc cagagaagct cgagcagcat gatgggcctg gtgacaatca
     1261 cgctaacttg gaaggtggct ccatcacctc tactgtctgg agacagcaaa gagcagaaag
     1321 cccactccgt gtccactggt gcctggagga ggctaaccca aggctggcac ctgtctccat
     1381 gacgacatga attcatacat atgcacagtg gtaccttaaa atttggataa cttcgtataa
     1441 tgtatgctat acgaagttat tcccaactca tcccggcctc gccggggccc cctagaaaaa
     1501 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1561 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1621 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1681 atgtctggat cccccgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     1741 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     1801 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     1861 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     1921 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     1981 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     2041 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     2101 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     2161 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     2221 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     2281 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     2341 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     2401 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     2461 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt cagccatggt tgtggcaagc
     2521 ttatcatcgt gtttttcaaa ggaaaaccac gtccccgtgg ttcggggggc ctagacgttt
     2581 ttttaacctc gactaaacac atgtaaagca tgtgcaccga ggccccagat cagatcccat
     2641 acaatggggt accttctggg catccttcag ccccttgttg aatacgcttg aggagagcca
     2701 tttgactctt tccacaacta tccaactcac aacgtggcac tggggttgtg ccgcctttgc
     2761 aggtgtatct tatacacgtg gcttttggcc gcagaggcac ctgtcgccag gtggggggtt
     2821 ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt caagaagctt ccagaggaac
     2881 tgcttccttc acgacattca acagaccttg cattcctttg gcgagagggg aaagacccct
     2941 aggaatgctc gtcaagaaga cagggccagg tttccgggcc ctcacattgc caaaagacgg
     3001 caatatggtg gaaaataaca tatagacaaa cgcacaccgg ccttattcca agcggcttcg
     3061 gccagtaacg ttctcgacgc tcgacctgca gttggacctg ggagtggaca cctgtggaga
     3121 gaaaggcaaa gtggatgtca ttgtcactca agtgtatggc cagatctcaa gcctgccaca
     3181 cctcaagctt gacaacaaaa agattgtctt ttctgaccag atggacgcgg ccaccctcaa
     3241 aggcatcacc gcgggccagg tgaatatcaa atcctcctcg tttttggaaa ctgacaatct
     3301 tagcgcagaa gtcatgcccg cttttgagag ggagtactca ccccaacagc tggccctcgc
     3361 agacagcgaa ttaattccat aacttcgtat aatgtatgct atacgaagtt atggatccac
     3421 tagttctaga ggccaccccc tctggaccca tgtctatcag agggggtagg gaagatggca
     3481 gggcagcaaa tatcaggcac agccaacaca caaacatcaa aaactgattt ttaaagtttc
     3541 tctaatgata aaaacaagga gtggtggtct cagaccttcc actggaagtc gatcacctgc
     3601 ctcactatta tttaaataaa agaacacact tatgaacaca aatggaaaat acaactacga
     3661 gagaaaaaat gacttgctta atatgtagaa attaactgtc tttgaaaaga acatgaagtt
     3721 tttataattt acatgaaaaa aaggcaaaca aacctggcta aacgtcggtt tattgtgcaa
     3781 ccgaaagcac ctgtctccat gacgacatgc tccaattttg aaataaaatg aacagttgac
     3841 tctgtaaggg aaaatgagag ctgattattt tgctgggaag atatcaaaca catggaatat
     3901 gtcagcagca tgacatacac tatcaaatta cttttaacct cacagaaaca tgacacagac
     3961 ttttttccgt ggtattaagc acctcctctt ccttgctgta ttttaggaca acgtaacttc
     4021 aacttccctt tctgcaaaca cactacttgt tcagcaaaga cggttctaac tctaacccgc
     4081 ctcctcctca ttcaaatccc ctccactttc ctaactgcac agctgttctg tttgtttctt
     4141 aaatggagta ttaaattagt ctaaaactaa atgtaactta ggcagtatct gttctccagt
     4201 tctccacgac caagttctct cttgaaaaca ccggtgtggt gagggttaag atcccaactc
     4261 tgctaccagc tgcttgggct tgggcaagtc accctagctc tcagatgtca tctgtaaatg
     4321 atgacaatgc caatgtggca ctgttctgag agtcagacag aacgtatgtg tgcttcacat
     4381 atggtgctca tgaagtgcta tcattatcta aggaaaacag aaaacgaagt tcagagtctc
     4441 tctaaacgca tgacaccaga ccaacgcgtt gatcaggcgc gccaccggtg cggccgcagg
     4501 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4561 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4621 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4681 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4741 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     4801 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4861 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4921 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4981 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5041 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     5101 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     5161 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5221 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5281 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5341 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5401 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5461 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5521 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5581 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5641 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5701 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5761 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5821 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5881 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5941 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6001 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6061 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6121 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6181 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6241 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6301 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6361 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6421 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6481 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6541 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6601 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6661 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6721 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6781 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6841 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6901 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6961 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa a
//
