LOCUS       sequence-464736-        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express BirA tagged to the C-terminus of CHIP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  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 7292)
  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-CHIP-BirA-HA
FEATURES             Location/Qualifiers
     source          1..7292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(272..1063,1064..1132))
                     /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"
     primer_bind     895..914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1133..1237)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1312..1331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1503..1882
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1883..2086
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2037..2057
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2131..2150
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2131..2149
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3131..4090
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             4094..4120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4151..4168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4157..4381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4427..4855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4514..4533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4724..4745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4864..4884)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4869..5198
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5049..5184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5111..5130
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5265..6059
                     /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
                     GLAPAELFARLKARMPNGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5319..5338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5929..5948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6233..6354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6270..6289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6324..6343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6403..6419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6403..6419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6416..6438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6427..6443
                     /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(6451..6457,6458..6475,6476..6481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6496..6517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6634..6651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6805..7292,1..101))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6885..6904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
       61 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
      121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
      181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
      241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
      301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
      361 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      421 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      541 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     1081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     1141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1201 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1261 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     1321 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     1381 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     1441 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     1501 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1561 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1621 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1681 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1741 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1801 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1861 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     1921 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1981 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2041 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
     2101 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     2161 tggctagcgc ttaaggcctg ttaaccggtc gtacatgaag ggcaaggagg agaaggaggg
     2221 cggcgcacgg ctgggcgctg gcggcggaag ccccgagaag agcccgagcg cgcaggagct
     2281 caaggagcag ggcaatcgtc tgttcgtggg ccgaaagtac ccggaggcgg cggcctgcta
     2341 cggccgcgcg atcacccgga acccgctggt ggccgtgtat tacaccaacc gggccttgtg
     2401 ctacctgaag atgcagcagc acgagcaggc cctggccgac tgccggcgcg ccctggagct
     2461 ggacgggcag tctgtgaagg cgcacttctt cctggggcag tgccagctgg agatggagag
     2521 ctatgatgag gccatcgcca atctgcagcg agcttacagc ctggccaagg agcagcggct
     2581 gaacttcggg gacgacatcc ccagcgctct tcgaatcgcg aagaagaagc gctggaacag
     2641 cattgaggag cggcgcatcc accaggagag cgagctgcac tcctacctct ccaggctcat
     2701 tgccgcggag cgtgagaggg agctggaaga gtgccagcga aaccacgagg gtgatgagga
     2761 cgacagccac gtccgggccc agcaggcctg cattgaggcc aagcacgaca agtacatggc
     2821 ggacatggac gagctttttt ctcaggtgga tgagaagagg aagaagcgag acatccccga
     2881 ctacctgtgt ggcaagatca gctttgagct gatgcgggag ccgtgcatca cgcccagtgg
     2941 catcacctac gaccgcaagg acatcgagga gcacctgcag cgtgtgggtc attttgaccc
     3001 cgtgacccgg agccccctga cccaggaaca gctcatcccc aacttggcta tgaaggaggt
     3061 tattgacgca ttcatctctg agaatggctg ggtggaggac tacttagtct ccggattcga
     3121 attcggatcc aaggacaaca ccgtgcccct gaagctgatc gccctgctgg ccaacggcga
     3181 gttccactct ggcgagcagc tgggagagac cctgggaatg agcagagccg ccatcaacaa
     3241 gcacatccag acactgagag actggggagt ggacgtgttc accgtgcctg gcaagggcta
     3301 cagcctgcct gagcctatcc agctgctgaa cgccaagcag atcctgggac agctggatgg
     3361 cggaagcgtg gccgtgctgc ctgtgatcga ctccaccaat cagtacctgc tggacagaat
     3421 cggagagctg aagtccggcg acgcctgcat cgccgagtac cagcaggctg gcagaggacg
     3481 cagaggacgg aagtggttca gcccattcgg agccaacctg tacctgtcca tgttctggag
     3541 actggagcag ggacctgctg ctgccatcgg actgagtctg gtgatcggaa tcgtgatggc
     3601 cgaggtgctg agaaagctgg gagccgacaa ggtgagagtg aagtggccta atgacctgta
     3661 cctccaggac cgcaagctgg ctggcatcct ggtggagctg acaggcaaga caggcgatgc
     3721 cgctcagatc gtgatcggag ccggaatcaa catggccatg agaagagtgg aggagagcgt
     3781 ggtgaaccag ggctggatca ccctgcagga ggctggcatc aacctggacc ggaacaccct
     3841 ggccgccatg ctgatcagag agctgagagc cgctctggag ctgttcgagc aggagggact
     3901 ggctccttac ctgagcagat gggagaagct ggacaacttc atcaacagac ctgtgaagct
     3961 gatcatcggc gacaaggaaa tcttcggcat ctccagagga atcgacaagc agggagctct
     4021 gctgctggag caggacggaa tcatcaagcc ctggatgggc ggagaaatct ccctgagaag
     4081 cgcagagaag gcttaccctt acgatgtacc ggattacgca taggcggcct aagtttaaac
     4141 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4201 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4261 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4321 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4381 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4441 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4501 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4561 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4621 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4681 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4741 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4801 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4861 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4921 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5041 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5101 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5161 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5221 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5281 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5341 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5401 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5461 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5521 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5581 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5641 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5701 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5761 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc caacggcgag gatctcgtcg
     5821 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5881 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5941 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6001 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6061 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6121 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6181 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6241 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6301 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6361 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6421 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6481 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7261 ctgcgctctg ctgaagccag ttaccttcgg aa
//
