LOCUS       sequence-435118-        5096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-TurboID fusion for proximity labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5096)
  TITLE     Expresses Flag-TurboID fusion for proximity labeling
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5096)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Flag-TurboID
FEATURES             Location/Qualifiers
     source          1..5096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1069..1092
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1099..2055
                     /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"
     polyA_signal    2126..2260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2150..2169)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2204..2223
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2373..2393)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2373..2391)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2412..2428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2412..2428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2425..2447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2436..2452
                     /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(2460..2466,2467..2484,2485..2490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2505..2526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2643..2660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2814..3402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2894..2913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3573..4364,4365..4433))
                     /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(4434..4538)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4564..5019)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4701..4722)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4913..4932
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccaccatgga ttacaaggat
     1081 gacgacgata agatccgcaa agacaatact gtgcctctga agctgatcgc tctcctggct
     1141 aatggcgagt tccatagtgg cgaacagctg ggagaaaccc tgggcatgtc cagggccgct
     1201 atcaacaagc acattcagac tctgcgcgac tggggcgtgg acgtgttcac cgtgcccgga
     1261 aagggctact ctctgcccga gcctatcccg ctgctgaacg ctaaacagat tctgggacag
     1321 ctggacggcg ggagcgtggc agtcctgcct gtggtcgact ccaccaatca gtacctgctg
     1381 gatcgaatcg gcgagctgaa gagtggggat gcttgcattg cagaatatca gcaggcaggg
     1441 agaggaagca gagggaggaa atggttctct ccttttggag ctaacctgta cctgagtatg
     1501 ttttggcgcc tgaagcgggg accagcagca atcggcctgg gcccggtcat cggaattgtc
     1561 atggcagaag cgctgcgaaa gctgggagca gacaaggtgc gagtcaaatg gcccaatgac
     1621 ctgtatctgc aggatagaaa gctggcaggc atcctggtgg agctggccgg aataacaggc
     1681 gatgctgcac agatcgtcat tggcgccggg attaacgtgg ctatgaggcg cgtggaggaa
     1741 agcgtggtca atcagggctg gatcacactg caggaagcag ggattaacct ggacaggaat
     1801 actctggccg ctacgctgat ccgagagctg cgggcagccc tggaactgtt cgagcaggaa
     1861 ggcctggctc catatctgcc acggtgggag aagctggata acttcatcaa tagacccgtg
     1921 aagctgatca ttggggacaa agagattttc gggattagcc gggggattga taaacaggga
     1981 gccctgctgc tggaacagga cggagttatc aaaccctgga tgggcggaga aatcagtctg
     2041 cggtctgccg aaaagctgca gggatctcag atctcgaatt caaggcctct cgagcctcta
     2101 gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg
     2161 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2221 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2281 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc
     2341 aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     2401 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2461 aacatacgag ccggragcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2521 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2581 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2641 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2701 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2761 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2821 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2881 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2941 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3001 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3061 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3121 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3181 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3241 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3301 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3361 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3421 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3481 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3541 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3601 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3661 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3721 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3781 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3841 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3901 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3961 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4021 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4081 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4141 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4201 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4261 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4321 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4381 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4441 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4501 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4561 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4621 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4681 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4741 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4801 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4861 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4921 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4981 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     5041 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgacc
//
