LOCUS       sequence-447896-        4477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mxiE-turboID fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4477)
  AUTHORS   Haidar-Ahmad N, Tomaro K, Lavallee-Adam M, Campbell-Valois F-X
  TITLE     The promiscuous biotin ligase TurboID reveals the proxisome of the 
            T3SS chaperone IpgC in Shigella flexneri.
  JOURNAL   mSphere. 2024 Nov 21;9(11):e0055324. doi: 10.1128/msphere.00553-24. 
            Epub 2024 Oct 31.
  PUBMED    39480076
REFERENCE   2  (bases 1 to 4477)
  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: "mSphere."; 
            date: "2024-11-21"; volume: "9(11):e0055324. doi"; pages: " 
            10.1128/msphere.00553-24. Epub 2024 Oct 31"
COMMENT     Sequence Label: pNHA13
FEATURES             Location/Qualifiers
     source          1..4477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3050..4006
                     /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"
     CDS             4013..4042
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4083..4100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4083..4099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4308..4327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4427..4449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcgattttt tcgcatattt ttcttgcaaa gttgggttga
     2161 gctggctaga ttagccagcc aatcttttgt atgtctgtgc gtacacagga aacagctatg
     2221 accatgtaac tagctaactc gagtaactag ctaagattac gaattcatgg gatcaagtaa
     2281 atataaaggt ctaaatacaa gtaatatgtt ttacatttac tctagtggac atgaaccagt
     2341 taacgttgag cttgtaaaag ataaagaacg taacataatt gaactggctc cagcatggaa
     2401 aggcttcttc ttcgtccgaa atcaaaatat aaaattcagt gataatgtga actatcatta
     2461 tcgctttaac attaatagtt gtgcaaaatt tcttgctttt tgggactatt tcagtggtgc
     2521 tctagtagag catagccatg ctgaaaaatg tattcatttc tatcatgaga atgatcttcg
     2581 ggatagttgt aataccgaat cgatgcttga taagttaatg ttaagattca tcttttcatc
     2641 tgaccagaat gtaagtaatg cactggctat gatacgaatg actgagtcat atcatttggt
     2701 tctttatctt ctgagaacta ttgagaaaga aaaggaggta agaataaaga gtctgacaga
     2761 gcattatggt gtgtcggagg cttattttag atctttatgt agaaaagctt taggggcaaa
     2821 agtcaaagaa caactgaaca cttggcggtt agtaaatggt ttattagatg tatttttgca
     2881 caatcaaaca atcacttcag ctgccatgaa taatgggtat gcatcaacat cccatttttc
     2941 aaatgagata aaaacacgac taggcttttc tgcaagagaa ttatcgaata ttactttttt
     3001 agtgaaaaaa ataaatgaaa aaattgggct cgagggatcc ggcggtggca aagacaatac
     3061 tgtgcctctg aagctgatcg ctctcctggc taatggcgag ttccatagtg gcgaacagct
     3121 gggagaaacc ctgggcatgt ccagggccgc tatcaacaag cacattcaga ctctgcgcga
     3181 ctggggcgtg gacgtgttca ccgtgcccgg aaagggctac tctctgcccg agcctatccc
     3241 gctgctgaac gctaaacaga ttctgggaca gctggacggc gggagcgtgg cagtcctgcc
     3301 tgtggtcgac tccaccaatc agtacctgct ggatcgaatc ggcgagctga agagtgggga
     3361 tgcttgcatt gcagaatatc agcaggcagg gagaggaagc agagggagga aatggttctc
     3421 tccttttgga gctaacctgt acctgagtat gttttggcgc ctgaagcggg gaccagcagc
     3481 aatcggcctg ggcccggtca tcggaattgt catggcagaa gcgctgcgaa agctgggagc
     3541 agacaaggtg cgagtcaaat ggcccaatga cctgtatctg caggatagaa agctggcagg
     3601 catcctggtg gagctggccg gaataacagg cgatgctgca cagatcgtca ttggcgccgg
     3661 gattaacgtg gctatgaggc gcgtggagga aagcgtggtc aatcagggct ggatcacact
     3721 gcaggaagca gggattaacc tggacaggaa tactctggcc gctacgctga tccgagagct
     3781 gcgggcagcc ctggaactgt tcgagcagga aggcctggct ccatatctgc cacggtggga
     3841 gaagctggat aacttcatca atagacccgt gaagctgatc attggggaca aagagatttt
     3901 cgggattagc cgggggattg ataaacaggg agccctgctg ctggaacagg acggagttat
     3961 caaaccctgg atgggcggag aaatcagtct gcggtctgcc gaaaagtccg gagaacagaa
     4021 gcttattagc gaagaggatc tgtaactaat ctagagtcga cctgcaggca tgcaagcttg
     4081 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4141 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4201 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     4261 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4321 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4381 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4441 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
