LOCUS       sequence-163083-        7039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rAAV-based template for genome engineering of the IQGAP1 C-terminus 
            containing PQS1 and 3xFLAG tags and a selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7039)
  AUTHORS   Vandemoortele G, Staes A, Gonnelli G, Samyn N, De Sutter D, 
            Vandermarliere E, Timmerman E, Gevaert K, Martens L, Eyckerman S
  TITLE     An extra dimension in protein tagging by quantifying universal 
            proteotypic peptides using targeted proteomics.
  JOURNAL   Sci Rep. 2016 Jun 6;6:27220. doi: 10.1038/srep27220.
  PUBMED    27264994
REFERENCE   2  (bases 1 to 7039)
  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: "Sci Rep."; 
            date: "2016-06-6"; volume: "6:27220. doi"; pages: " 
            10.1038/srep27220"
COMMENT     Sequence Label: pAav-IQGAP1-PQS1-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7039
                     /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
     protein_bind    1180..1213
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     CDS             1284..1349
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1377..1410)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1422..1922
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1457..1480)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1855..1874
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1936..2739
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVNQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1999..2018)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2609..2628
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2771..2788)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2777..3001
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(3026..3059)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   4302..4442
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4302..4431
                     /label=AAV2 ITR
     rep_origin      4517..4972
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4604..4623)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4814..4835
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4989..5008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5108..5130
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5168..5186)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5254..5358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5359..5427,5428..6219)
                     /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(5577..5596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6390..6978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6879..6898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca aacgcgtaaa gggccccttg tggggtcctt
      181 tcaagacttt tgacccttgc tttctcatca ccactcttat aaagttcaga agtgagacca
      241 ctcagatttc ttaaacttgt tcaggacaat ggtgaaagta gcaggaaaag cgggagcatt
      301 gcagtcagat ctggatttat cgcctccatc tctgtcctgt tgtgcaagtc atttaacatc
      361 tctgagtctc agtagctcat ctttaaagtt agaataatcg cacctcataa aaatttttgt
      421 gaggacttta aaagatacta tgtaatgtgc ttacacaatg ccaagcacat ggcagacacc
      481 taataaaaca tcaagttctc tcttttcccc ttgccttcgg gatagaaagt gttctcatat
      541 ggaatagaaa ttaagaatct atcagatagt attaaggtga cattactagc cttatctcat
      601 ttccccgtgc ttagtatatt tcatcatagg tttaaatgct tcaccttttc cagatttatt
      661 aaaactagtg acccataaca tggatgaaat gggaaaatgt cattgtgact tctgaaataa
      721 aaagcttaga tgtgtgtggc aattgaacat gtttcttttt ctttccatgt atgatgaatt
      781 ttgatatgaa caacatgact tcctaactcc aaatcaacca gaagcctccc ttggccaatg
      841 tgccctattg ctccagctct caggctggtt gtgcagtggg tagttgccta gcgtggaact
      901 tttctttcgc cccagttcac atctggcaca caaggcaggc ctcagtccat ctggatccct
      961 ccctacattg tgtccttttt ccttgttcca cagacttgat taactgtcaa aatgttttgt
     1021 tttgttttgt tttgttttgt taataggacc tgctgcagct acagtatgaa ggagttgcag
     1081 tcatgaaatt atttgataga gctaaagtaa atgtcaacct cctgatcttc cttctcaaca
     1141 aaaagttcta cgggaaggaa ttcgggaagt tcctattccg aagttcctat tcttcaaata
     1201 gtataggaac ttcactgacc ctgatcttca gagaggccgt gagcgaaatc ctggagacct
     1261 ccagattcgc ctacctgtac gacgactaca aagaccatga cggtgattat aaagatcatg
     1321 atattgatta caaggatgac gatgacaagg gcgagcccga ggcttgaaag cttcggataa
     1381 cttcgtatag catacattat acgaagttat cgcgccctac cgggtagggg aggcgctttt
     1441 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     1501 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     1561 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     1621 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     1681 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     1741 gctttggctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
     1801 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaagc ccggcattct
     1861 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     1921 cgacctgcag ccaatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
     1981 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2041 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2101 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2161 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2221 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2281 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2341 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2401 gtcaatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2461 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2521 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2581 ggtgtggcgg atcgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2641 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     2701 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag gggatcaatt ctctagagct
     2761 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2821 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2881 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2941 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3001 gcttctgagg cggaaagaac cagctataac ttcgtatagc atacattata cgaagttata
     3061 gtaatcgata actcgagtaa ttgatcgttt gctgccagcc cagaaggatg aaggaaagaa
     3121 gcacctcaca gctcctttct aggtccttct ttcctcattg gaagcaaaga cctagccaac
     3181 aacagcacct caatctgata cactcccgat gccacatttt taactcctct cgctctgatg
     3241 ggacatttgt tacccttttt tcatagtgaa attgtgtttc aggcttagtc tgacctttct
     3301 ggtttcttca ttttcttcca ttacttagga aagagtggaa actccactaa aatttctctg
     3361 tgttgttaca gtcttagagg ttgcagtact atattgtaag ctttggtgtt tgtttaatta
     3421 gcaataggga tggtaggatt caaatgtgtg tcatttagaa gtggaagcta ttagcaccaa
     3481 tgacataaat acatacaaga cacacaacta aaatgtcatg ttattaacag ttattaggtt
     3541 gtcatttaaa aataaagttc ctttatattt ctgtcccatc aggaaaactg aaggatatgg
     3601 ggaatcattg gttatcttcc attgtgtttt tctttatgga caggagctaa tggaagtgac
     3661 agtcatgttc aaaggaagca tttctagaaa aaaggagata atgtttttaa atttcattat
     3721 caaacttggg caattctgtt tgtgtaactc cccgactagt ggatgggaga gtcccattgc
     3781 taaaattcag ctactcagat aaattcagaa tgggtcaagg cacctgcctg tttttgttgg
     3841 tgcacagaga ttgacttgat tcagagagac aattcactcc atccctatgg cagaggaatg
     3901 ggttagccct aatgtagaat gtcattgttt ttaaaactgt tttatatctt aagagtgcct
     3961 tattaaagta tagatgtatg tcttaaaatg tgggtgatag gaattttaaa gatttatata
     4021 atgcatcaaa agccttagaa taagaaaagc tttttttaaa ttgctttatc tgtatatctg
     4081 aactcttgaa acttatagct aaaacactag gatttatctg cagtgttcag ggagataatt
     4141 ctgcctttaa ttgtctaaaa caaaaacaaa accagccaac ctatgttaca cgtgagatta
     4201 aaaccaattt tttccccatt ttttctcctt ttttctcttg ctgcccacat tgtgccttta
     4261 ttttatgagc cccagttttc tgggctagca aaagcggccg caggaacccc tagtgatgga
     4321 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4381 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     4441 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4501 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4561 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4621 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4681 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     4741 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4801 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     4861 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4921 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4981 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5041 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5101 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5161 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5221 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5341 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5401 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5461 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5521 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5581 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5641 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5701 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5761 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5821 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5881 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5941 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6001 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6061 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6121 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     6541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7021 tggccttttg ctcacatgt
//
