LOCUS       sequence-477848-        4394 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  Gateway entry clone with ER-localized TurboID tagged with 
            superfolder GFP (contains stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4394)
  TITLE     Perrimon Lab Plasmids 2025
REFERENCE   2  (bases 1 to 4394)
  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_BIP-sfGFP-TurboID-KDEL
FEATURES             Location/Qualifiers
     source          1..4394
                     /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     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"
     sig_peptide     689..742
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1529..2485
                     /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(2519..2618)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2635..2654)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2636..2654)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2659..2675)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2659..2675)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2777..2796)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2788..3597
                     /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(2855..2874)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3744..4332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4233..4252
                     /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 gaagttatgc atattactgg ccgtcgtggc
      721 ctttgttggc ctctcgctcg gggaatccaa gggcgaggag ctgttcaccg gcgtggtgcc
      781 catcctggtg gagctggatg gcgacgtgaa cggccacaag ttcagcgtgc gcggcgaggg
      841 cgagggcgac gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      901 gcccgtgccc tggcccaccc tggtgaccac cctgacctac ggcgtgcagt gcttcagccg
      961 ctaccccgat cacatgaagc agcacgattt cttcaagagc gccatgcccg agggctacgt
     1021 gcaggagcgc accatcagct tcaaggatga cggcacctac aagacccgcg ccgaggtgaa
     1081 gttcgagggc gataccctgg tgaaccgcat cgagctgaag ggcatcgatt tcaaggagga
     1141 tggcaacatc ctgggccaca agctggagta caacttcaac agccacaacg tgtacatcac
     1201 cgccgataag cagaagaacg gcatcaaggc caacttcaag atccgccaca atgtggagga
     1261 tggctccgtg cagctggccg atcactacca gcagaacacc cccatcggcg acggcccagt
     1321 gctgctgccc gataaccact acctgagcac ccagagcgtg ctgtccaagg accccaacga
     1381 gaagcgcgat cacatggtgc tgctggagtt cgtgaccgcc gccggcatca ccctgggcat
     1441 ggatgagctg tacaagaccg gtggttcagg cggttctggt ggtagcggcg gtagcggtgg
     1501 tagcggtggt agcggtggca gcggcggtaa agacaatact gtgcctctga agctgatcgc
     1561 tctcctggct aatggcgagt tccatagtgg cgaacagctg ggagaaaccc tgggcatgtc
     1621 cagggccgct atcaacaagc acattcagac tctgcgcgac tggggcgtgg acgtgttcac
     1681 cgtgcccgga aagggctact ctctgcccga gcctatcccg ctgctgaacg ctaaacagat
     1741 tctgggacag ctggacggcg ggagcgtggc agtcctgcct gtggtcgact ccaccaatca
     1801 gtacctgctg gatcgaatcg gcgagctgaa gagtggggat gcttgcattg cagaatatca
     1861 gcaggcaggg agaggaagca gagggaggaa atggttctct ccttttggag ctaacctgta
     1921 cctgagtatg ttttggcgcc tgaagcgggg accagcagca atcggcctgg gcccggtcat
     1981 cggaattgtc atggcagaag cgctgcgaaa gctgggagca gacaaggtgc gagtcaaatg
     2041 gcccaatgac ctgtatctgc aggatagaaa gctggcaggc atcctggtgg agctggccgg
     2101 aataacaggc gatgctgcac agatcgtcat tggcgccggg attaacgtgg ctatgaggcg
     2161 cgtggaggaa agcgtggtca atcagggctg gatcacactg caggaagcag ggattaacct
     2221 ggacaggaat actctggccg ctacgctgat ccgagagctg cgggcagccc tggaactgtt
     2281 cgagcaggaa ggcctggctc catatctgcc acggtgggag aagctggata acttcatcaa
     2341 tagacccgtg aagctgatca ttggggacaa agagattttc gggattagcc gggggattga
     2401 taaacaggga gccctgctgc tggaacagga cggagttatc aaaccctgga tgggcggaga
     2461 aatcagtctg cggtctgccg aaaagaagga cgagctgtaa tagaagggtg gggcgccgac
     2521 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2581 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     2641 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2701 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2761 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2821 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2881 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2941 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     3001 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     3061 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     3121 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     3181 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     3241 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     3301 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     3361 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     3421 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     3481 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     3541 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     3601 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     3661 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4381 tttgctcaca tgtt
//
