LOCUS       sequence-413768-        4048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty C-terminus donor cassette. Integrate homology arms to target 
            the C-terminus insertion of a moxGFP-2A-Blast cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4048)
  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 4048)
  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-moxGFP-Blast
FEATURES             Location/Qualifiers
     source          1..4048
                     /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             join(444..446,447..449,450..1160)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(489..510)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1097..1118
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1176..1217
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(1239..1272)
                     /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)."
     CDS             1284..1340
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1347..1742
                     /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(1749..1782)
                     /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(1827..1843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1827..1843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1840..1862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1851..1867
                     /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(1875..1881,1882..1899,1900..1905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1920..1941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2058..2075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2229..2817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2309..2328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2988..3779,3780..3848))
                     /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     3611..3630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3849..3953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4021..4039
                     /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 ggagctgttc accggggtgg
      481 tgcccatcct ggtcgagctg gacggcgacg tgaacggcca caagttctcc gtgcggggcg
      541 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatcagc accaccggca
      601 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagagcttct
      661 cccgctaccc cgaccacatg aagcgccacg acttcttcaa gagcgccatg cccgaaggct
      721 acgtccagga gcgcaccatc tccttcaagg acgacggcac ctacaagacc cgcgccgagg
      781 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      841 aggacggcaa catcctgggg cacaagctgg agtacaactt caactcccac aacgtctata
      901 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cataacgtgg
      961 aggacggctc cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1021 ccgtgctgct gcccgacaac cactacctgt ccacccagtc caagctgtcc aaagacccca
     1081 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactcacg
     1141 gcatggacga gctgtacaag ggaggtggcg gttccggcaa gcccatcccc aaccccctgc
     1201 tgggcctgga cagcaccgga ggtggcggat caacgcgtta ccgttcgtat agcatacatt
     1261 atacgaagtt atctggatcc ggagctacta acttcagcct gctgaagcag gccggagacg
     1321 tggaggagaa ccctggacct tctagaatgg ccaagccttt gtctcaagaa gaatccaccc
     1381 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     1441 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     1501 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     1561 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     1621 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     1681 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     1741 gccccgggat aacttcgtat agcatacatt atacgaacgg tagcctcgag accggtaaga
     1801 gcaagtgaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1861 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1921 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1981 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2041 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2101 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2161 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2221 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2281 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2341 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2401 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2461 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2521 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2581 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2641 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2701 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2761 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2821 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2881 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2941 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3001 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3061 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3121 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3181 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3241 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3301 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3361 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3421 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3481 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3541 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3601 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3661 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3721 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3781 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3841 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3901 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3961 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4021 aataggcgta tcacgaggcc ctttcgtc
//
