LOCUS       sequence-413771-        4108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty C-terminus donor cassette. Integrate homology arms to target 
            the C-terminus insertion of a miniTurbo-2A-Blast cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4108)
  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 4108)
  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-miniTurbo-Blast
FEATURES             Location/Qualifiers
     source          1..4108
                     /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             450..1220
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             1236..1277
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(1299..1332)
                     /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             1344..1400
                     /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             1407..1802
                     /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(1809..1842)
                     /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(1887..1903)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1887..1903)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1900..1922)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1911..1927
                     /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(1935..1941,1942..1959,1960..1965))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1980..2001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2118..2135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2289..2877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2369..2388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3048..3839,3840..3908))
                     /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"
     promoter        complement(3909..4013)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4081..4099
                     /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 tctatggcga tcccgctgct gaacgctaaa cagattctgg
      481 gacagctgga cggcgggagc gtggcagtcc tgcctgtggt cgactccacc aatcagtacc
      541 tgctggatcg aatcggcgag ctgaagagtg gggatgcttg cattgcagaa tatcagcagg
      601 cagggagagg aagcagaggg aggaaatggt tctctccttt tggagctaac ctgtacctga
      661 gtatgttttg gcgcctgaag cggggaccag cagcaatcgg cctgggcccg gtcatcggaa
      721 ttgtcatggc agaagcgctg cgaaagctgg gagcagacaa ggtgcgagtc aaatggccca
      781 atgacctgta tctgcaggat agaaagctgg caggcatcct ggtggagctg gccggaataa
      841 caggcgatgc tgcacagatc gtcattggcg ccgggattaa cgtggctatg aggcgcgtgg
      901 aggaaagcgt ggtcaatcag ggctggatca cactgcagga agcagggatt aacctggaca
      961 ggaatactct ggccgctatg ctgatccgag agctgcgggc agccctggaa ctgttcgagc
     1021 aggaaggcct ggctccatat ctgtcacggt gggagaagct ggataacttc atcaatagac
     1081 ccgtgaagct gatcattggg gacaaagaga ttttcgggat tagccggggg attgataaac
     1141 agggagccct gctgctggaa caggacggag ttatcaaacc ctggatgggc ggagaaatca
     1201 gtctgcggtc tgccgaaaag ggaggtggcg gttccggcaa gcccatcccc aaccccctgc
     1261 tgggcctgga cagcaccgga ggtggcggat caacgcgtta ccgttcgtat agcatacatt
     1321 atacgaagtt atctggatcc ggagctacta acttcagcct gctgaagcag gccggagacg
     1381 tggaggagaa ccctggacct tctagaatgg ccaagccttt gtctcaagaa gaatccaccc
     1441 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     1501 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     1561 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     1621 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     1681 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     1741 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     1801 gccccgggat aacttcgtat agcatacatt atacgaacgg tagcctcgag accggtaaga
     1861 gcaagtgaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1921 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1981 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2101 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4021 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4081 aataggcgta tcacgaggcc ctttcgtc
//
