LOCUS       sequence-393507-        6473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transiently expresses golgi localized TurboID with HA tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6473)
  AUTHORS   Yan T, Julio AR, Villanueva M, Jones AE, Ball AB, Boatner LM, Turmon
            AC, Nguyen KB, Yen SL, Desai HS, Divakaruni AS, Backus KM
  TITLE     Proximity-labeling chemoproteomics defines the subcellular 
            cysteinome and inflammation-responsive mitochondrial redoxome.
  JOURNAL   Cell Chem Biol. 2023 Jul 20;30(7):811-827.e7. doi: 
            10.1016/j.chembiol.2023.06.008. Epub 2023 Jul 6.
  PUBMED    37419112
REFERENCE   2  (bases 1 to 6473)
  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"; doi: 
            "10.1016/j.chembiol.2023.06.008"; journalName: "Cell Chem Biol"; 
            date: "2023-07-20- 20"; volume: "30"; issue: "7"; pages: 
            "811-827.e7"
COMMENT     Sequence Label: Golgi-TurboID
FEATURES             Location/Qualifiers
     source          1..6473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        129..508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        509..712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     663..683
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     757..776
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             804..830
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             834..860
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             864..890
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             897..1853
                     /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"
     promoter        complement(1920..1938)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1921..1938)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1958..1975)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1964..2188
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2234..2662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2321..2340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2531..2552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2671..2691)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2676..3005
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2856..2991
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2918..2937
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3072..3866
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3126..3145)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3736..3755
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4040..4161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4131..4150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4210..4226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4210..4226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4223..4245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4234..4250
                     /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(4258..4264,4265..4282,4283..4288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4303..4324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4441..4458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4612..5200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4692..4711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5371..6162,6163..6231))
                     /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(6232..6336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6411..6430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
       61 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      121 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      481 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      541 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      601 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      661 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      721 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      781 ccaagcttgc ggccgccacc atgtacccgt atgatgttcc ggattacgct ggctatccct
      841 acgacgtgcc cgactatgcc gggtacccct atgacgtccc agactacgca gctagcaaag
      901 acaatactgt gcctctgaag ctgatcgctc tcctggctaa tggcgagttc catagtggcg
      961 aacagctggg agaaaccctg ggcatgtcca gggccgctat caacaagcac attcagactc
     1021 tgcgcgactg gggcgtggac gtgttcaccg tgcccggaaa gggctactct ctgcccgagc
     1081 ctatcccgct gctgaacgct aaacagattc tgggacagct ggacggcggg agcgtggcag
     1141 tcctgcctgt ggtcgactcc accaatcagt acctgctgga tcgaatcggc gagctgaaga
     1201 gtggggatgc ttgcattgca gaatatcagc aggcagggag aggaagcaga gggaggaaat
     1261 ggttctctcc ttttggagct aacctgtacc tgagtatgtt ttggcgcctg aagcggggac
     1321 cagcagcaat cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg ctgcgaaagc
     1381 tgggagcaga caaggtgcga gtcaaatggc ccaatgacct gtatctgcag gatagaaagc
     1441 tggcaggcat cctggtggag ctggccggaa taacaggcga tgctgcacag atcgtcattg
     1501 gcgccgggat taacgtggct atgaggcgcg tggaggaaag cgtggtcaat cagggctgga
     1561 tcacactgca ggaagcaggg attaacctgg acaggaatac tctggccgct acgctgatcc
     1621 gagagctgcg ggcagccctg gaactgttcg agcaggaagg cctggctcca tatctgccac
     1681 ggtgggagaa gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag
     1741 agattttcgg gattagccgg gggattgata aacagggagc cctgctgctg gaacaggacg
     1801 gagttatcaa accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aaggaattct
     1861 tcctttggcg gattttctgt tttcggaagt gactcgagca tgcatctaga gggccctatt
     1921 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     1981 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2041 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2101 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2161 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2221 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2281 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2581 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2641 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2821 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2881 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2941 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3001 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3061 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3121 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3181 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3241 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3301 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3361 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3421 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3481 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3541 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3601 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3661 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3721 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3781 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3841 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3901 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3961 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4021 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4081 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4141 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6361 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     6421 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggt
//
