LOCUS       sequence-464922-        5052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HA-BirA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5052)
  AUTHORS   Mukhopadhyay U, Levantovsky S, Carusone TM, Gharbi S, Stein F, 
            Behrends C, Bhogaraju S
  TITLE     A ubiquitin-specific, proximity-based labeling approach for the 
            identification of ubiquitin ligase substrates.
  JOURNAL   Sci Adv. 2024 Aug 9;10(32):eadp3000. doi: 10.1126/sciadv.adp3000. 
            Epub 2024 Aug 9.
  PUBMED    39121224
REFERENCE   2  (bases 1 to 5052)
  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: "Sci Adv."; 
            date: "2024-08-9"; volume: "10(32):eadp3000. doi"; pages: " 
            10.1126/sciadv.adp3000. Epub 2024 Aug 9"
COMMENT     Sequence Label: pEHAC1-HA-BirAonly
FEATURES             Location/Qualifiers
     source          1..5052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(888..914)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(969..995)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1043..1246)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1072..1092)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1247..1550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1725..2313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1805..1824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2642..2689
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2714..2733
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2921..3715)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(2921..3715)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3032..3051)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3642..3661
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3750..4107)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3764..3899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3916..4107
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4064..4084
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4109..4213)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4240..4695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4537..4558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4702..4823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4713..4732)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4767..4786
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(4953..5052,1..866))
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             complement(join(4956..5052,1..866))
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
ORIGIN
        1 gaaatgccaa atatttcttt atcaccaatg ataagtttca ctgggcgatt aataaaatta
       61 tccagctttt cccagcgcga cagataaggt gccaatcctt cttgttcgaa gagttccaac
      121 gcagcacgta attcacgtat tagcatggcc gccaacgtat tacgatcgag attgatcccc
      181 gcttcctgca gcgtgatcca cccctgatta acgacactct cttcaacacg gcgcattgcc
      241 atgttgatcc cggctccaat gactatttgc gccgcatcgc cagttttgcc agtcagctcc
      301 accagaatgc ctgccagctt gcgatcctgc agatagaggt cattaggcca tttaacacga
      361 actttatctg cacccagctt gcgtaatact tccgccatca cgataccgat aaccagactt
      421 aaaccaatcg ccgccgccgg gccttgttcc agacgccaga acatcgacaa atataagttt
      481 gcgccaaaag gcgaaaacca tttccgaccc cggcgaccac ggccagcctg ctggtattct
      541 gcaatgcaag catcgcccga tttaagctct ccgatacgat caagaaggta ctgattcgtg
      601 gagtcaatca caggcagcac ggctacacta ccgccatcca gctgacccaa tatctgttta
      661 gcattaagta actggatagg ctcaggcagg ctgtatcctt tacccggaac ggtaaagaca
      721 tcaacgcccc agtcacgcag tgtctgaatg tgtttattaa tagccgcccg gctcattccc
      781 agcgtttcac ccaactgctc gccagagtga aattcaccgt tcgctaacag ggcaatcaat
      841 ttcagtggca cggtgttatc cttcatccct cgagatctga gtccggaagc gtagtctggg
      901 acgtcgtatg ggtacatggt accgtcgact gcagaattcg aagcttgagc tcgagatctg
      961 agtccggaag cgtagtctgg gacgtcgtat gggtacatgg tggcgaccgg tagcgctagc
     1021 ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     1081 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     1141 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     1201 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     1261 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     1321 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     1381 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     1441 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     1501 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     1561 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg
     1621 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2161 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2221 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2281 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2341 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2401 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2461 tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg
     2521 ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc
     2581 caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta
     2641 tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
     2701 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag
     2761 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     2821 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg
     2881 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     2941 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     3001 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     3061 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     3121 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     3181 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     3241 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     3301 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     3361 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     3421 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     3481 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt
     3541 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     3601 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     3661 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     3721 acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct
     3781 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     3841 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     3901 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     3961 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     4021 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     4081 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
     4141 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4201 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4261 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4321 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4381 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4441 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4501 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4561 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4621 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4681 taacgcttac aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa
     4741 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4801 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4861 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     4921 attatgatca gttatctaga tccggtggat ccttattttt ctgcactacg cagggatatt
     4981 tcaccgccca tccagggttt tattattcca tcctgctcaa gtaataaagc cccctgtttg
     5041 tctattccgc gt
//
