LOCUS       sequence-477851-        4329 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  Gateway entry clone with TurboID tagged with superfolder GFP 
            (contains stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4329)
  TITLE     Perrimon Lab Plasmids 2025
REFERENCE   2  (bases 1 to 4329)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pENTR_sfGFP-TurboID
FEATURES             Location/Qualifiers
     source          1..4329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1475..2431
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     protein_bind    complement(2454..2553)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2570..2589)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2571..2589)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2594..2610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2594..2610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2712..2731)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2723..3532
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2790..2809)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3679..4267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4168..4187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat gtccaagggc gaggagctgt tcaccggcgt
      721 ggtgcccatc ctggtggagc tggatggcga cgtgaacggc cacaagttca gcgtgcgcgg
      781 cgagggcgag ggcgacgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
      841 caagctgccc gtgccctggc ccaccctggt gaccaccctg acctacggcg tgcagtgctt
      901 cagccgctac cccgatcaca tgaagcagca cgatttcttc aagagcgcca tgcccgaggg
      961 ctacgtgcag gagcgcacca tcagcttcaa ggatgacggc acctacaaga cccgcgccga
     1021 ggtgaagttc gagggcgata ccctggtgaa ccgcatcgag ctgaagggca tcgatttcaa
     1081 ggaggatggc aacatcctgg gccacaagct ggagtacaac ttcaacagcc acaacgtgta
     1141 catcaccgcc gataagcaga agaacggcat caaggccaac ttcaagatcc gccacaatgt
     1201 ggaggatggc tccgtgcagc tggccgatca ctaccagcag aacaccccca tcggcgacgg
     1261 cccagtgctg ctgcccgata accactacct gagcacccag agcgtgctgt ccaaggaccc
     1321 caacgagaag cgcgatcaca tggtgctgct ggagttcgtg accgccgccg gcatcaccct
     1381 gggcatggat gagctgtaca agaccggtgg ttcaggcggt tctggtggta gcggcggtag
     1441 cggtggtagc ggtggtagcg gtggcagcgg cggtaaagac aatactgtgc ctctgaagct
     1501 gatcgctctc ctggctaatg gcgagttcca tagtggcgaa cagctgggag aaaccctggg
     1561 catgtccagg gccgctatca acaagcacat tcagactctg cgcgactggg gcgtggacgt
     1621 gttcaccgtg cccggaaagg gctactctct gcccgagcct atcccgctgc tgaacgctaa
     1681 acagattctg ggacagctgg acggcgggag cgtggcagtc ctgcctgtgg tcgactccac
     1741 caatcagtac ctgctggatc gaatcggcga gctgaagagt ggggatgctt gcattgcaga
     1801 atatcagcag gcagggagag gaagcagagg gaggaaatgg ttctctcctt ttggagctaa
     1861 cctgtacctg agtatgtttt ggcgcctgaa gcggggacca gcagcaatcg gcctgggccc
     1921 ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg ggagcagaca aggtgcgagt
     1981 caaatggccc aatgacctgt atctgcagga tagaaagctg gcaggcatcc tggtggagct
     2041 ggccggaata acaggcgatg ctgcacagat cgtcattggc gccgggatta acgtggctat
     2101 gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc acactgcagg aagcagggat
     2161 taacctggac aggaatactc tggccgctac gctgatccga gagctgcggg cagccctgga
     2221 actgttcgag caggaaggcc tggctccata tctgccacgg tgggagaagc tggataactt
     2281 catcaataga cccgtgaagc tgatcattgg ggacaaagag attttcggga ttagccgggg
     2341 gattgataaa cagggagccc tgctgctgga acaggacgga gttatcaaac cctggatggg
     2401 cggagaaatc agtctgcggt ctgccgaaaa gtaatagaag ggtgggcgcg ccgacccagc
     2461 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2521 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2581 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2641 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2701 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2761 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2821 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2881 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2941 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3001 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3061 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3121 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3181 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3241 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3301 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3361 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3421 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3481 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3541 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3601 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3661 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3721 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3781 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3841 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3901 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3961 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4021 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4081 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4141 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4201 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4261 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4321 tcacatgtt
//
