LOCUS       sequence-451272-        4467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty C-terminus donor cassette. Integrate homology arms to target 
            the C-terminus insertion of a mScarlet-EFS-Blast cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4467)
  AUTHORS   Philip R, Sharma A, Matellan L, Erpf AC, Hsu WH, Tkach JM, Wyatt 
            HDM, Pelletier L
  TITLE     qTAG: an adaptable plasmid scaffold for CRISPR-based endogenous 
            tagging.
  JOURNAL   EMBO J. 2024 Dec 12. doi: 10.1038/s44318-024-00337-5.
  PUBMED    39668248
REFERENCE   2  (bases 1 to 4467)
  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: "EMBO J. 
            2024 Dec 12. doi: 10.1038/s44318-024-00337-5."
COMMENT     Sequence Label: qTAG-C-mScarlet-EFS-Blast
FEATURES             Location/Qualifiers
     source          1..4467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             444..1139
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(577..595)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     845..864
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1155..1196
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(1218..1251)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1373..1494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1410..1429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1464..1483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1544..1755
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             1766..2161
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(2168..2201)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2246..2262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2246..2262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2259..2281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2270..2286
                     /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)."
     promoter        complement(join(2294..2300,2301..2318,2319..2324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2339..2360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2477..2494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2648..3236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2728..2747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3407..4198,4199..4267))
                     /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     4030..4049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4268..4372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4440..4458
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cggaggaagc ggaggatctg
      421 gcggtaccgg tggaggcgga tctatggtga gcaagggcga ggcagtgatc aaggagttca
      481 tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
      541 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      601 gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct
      661 tcaccaagca ccccgccgac atccccgact actataagca gtccttcccc gagggcttca
      721 agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc caggacacct
      781 ccctggagga cggcaccctg atctacaagg tgaagctccg cggcaccaac ttccctcctg
      841 acggccccgt aatgcagaag aagacaatgg gctgggaagc gtccaccgag cggttgtacc
      901 ccgaggacgg cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc
      961 gctacctggc ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg
     1021 cctacaacgt cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg
     1081 aacagtacga acgctccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg
     1141 gaggtggcgg ttccggcaag cccatcccca accccctgct gggcctggac agcaccggag
     1201 gtggcggatc aacgcgttac cgttcgtata gcatacatta tacgaagtta tctggatcct
     1261 gataatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1321 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttactagat
     1501 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     1561 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg
     1621 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     1681 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     1741 gccgccagaa cacagggcct ctagaatggc caagcctttg tctcaagaag aatccaccct
     1801 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     1861 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     1921 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     1981 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     2041 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     2101 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     2161 ccccgggata acttcgtata gcatacatta tacgaacggt agcctcgaga ccggtaagag
     2221 caagtgaaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2281 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2341 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2401 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2461 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2521 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2581 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2641 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2701 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2761 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2821 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2881 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2941 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3001 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3061 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3121 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3181 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3241 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3301 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3361 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3421 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3481 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3541 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3601 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3661 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3721 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3781 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3841 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3901 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3961 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4021 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4081 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4141 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4201 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4261 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4321 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4381 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4441 ataggcgtat cacgaggccc tttcgtc
//
