LOCUS       sequence-364143-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 ORF3a fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  AUTHORS   May DG, Martin-Sancho L, Anschau V, Liu S, Chrisopulos RJ, Scott KL,
            Halfmann CT, Diaz Pena R, Pratt D, Campos AR, Roux KJ
  TITLE     A BioID-Derived Proximity Interactome for SARS-CoV-2 Proteins.
  JOURNAL   Viruses. 2022 Mar 15;14(3). pii: v14030611. doi: 10.3390/v14030611.
  PUBMED    35337019
REFERENCE   2  (bases 1 to 6634)
  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: "Viruses."; 
            date: "2022-03-15"; volume: "14(3). pii: v14030611. doi"; pages: " 
            10.3390/v14030611"
COMMENT     Sequence Label: ORF3a-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2..471
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    536..735
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             936..1352
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1282..1304
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1320..1336
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1363..1372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2221..2916
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             2917..2943
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2958..2978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2963..3292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3143..3278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3302..3901
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3302..3321)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3798..3818
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4122..4591
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4721..4738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4892..5480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4972..4991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5640..6431,6432..6500))
                     /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     6263..6282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6501..6605)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
       61 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
      121 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
      181 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
      241 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
      301 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
      361 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
      421 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
      481 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggcaagct
      541 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc
      601 tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
      661 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
      721 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
      781 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
      841 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
      901 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
      961 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1021 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1081 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1141 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1201 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1261 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1321 tttatccagc cctcactcct tctctaggcg ccggccggat ccgccaccat ggaccttttt
     1381 atgagaatct ttaccatcgg gactgtcaca ctgaaacagg gagaaattaa ggacgcgacc
     1441 ccgagtgact ttgtgagggc cactgctaca attcccatcc aagcctctct tccattcggc
     1501 tggctcatag tgggtgtggc actgctcgcg gtattccagt cagcgtctaa gatcataacg
     1561 ctgaagaaga gatggcagtt ggctctcagc aaaggtgttc atttcgtctg taacctgctt
     1621 cttcttttcg tcactgtcta ttctcacttg ctcttggttg cagccgggct ggaagccccg
     1681 tttctgtatc tgtacgcgct cgtttacttc ctccagagca taaatttcgt acgaattatt
     1741 atgagactct ggttgtgctg gaagtgtcgg agcaagaatc ctctcttgta tgacgccaat
     1801 tacttccttt gttggcacac taattgttat gattactgca taccttacaa tagcgttacc
     1861 tcatccatag taatcacctc cggtgacgga acaacgtcac caatctccga gcatgattac
     1921 cagattggag gctatacgga aaaatgggag agtggtgtca aggactgtgt ggtgctgcat
     1981 agctatttca caagcgatta ttatcaactg tacagtacac agttgtccac ggataccggg
     2041 gtcgagcatg tcacattttt tatttataac aagatcgttg atgagccaga ggagcatgtg
     2101 cagatccaca cgatagatgg aagctccggt gtagtgaatc cagtcatgga accgatctat
     2161 gacgagccca ccacgaccac atctgtgccc ctgaccggtg gtggaggcgg gtctgaattc
     2221 ttcaagaacc tgatctggct gaaggaggtg gacagcaccc aggagagact gaaggagtgg
     2281 aacgtgagct acggcaccgc cctggtggcc gacagacaga ccaagggcag aggcggcctg
     2341 ggcagaaagt ggctgagcca ggagggcggc ctgtacttca gcttcctgct gaaccccaag
     2401 gagttcgaga acctgctgca gctgcccctg gtgctgggcc tgagcgtgag cgaggccctg
     2461 gaggagatca ccgagatccc cttcagcctg aagtggccca acgacgtgta cttccaggag
     2521 aagaaggtga gcggcgtgct gtgcgagctg agcaaggaca agctgatcgt gggcatcggc
     2581 atcaacgtga accagagaga gatccccgag gagatcaagg acagagccac caccctgtac
     2641 gagatcaccg gcaaggactg ggacagaaag gaggtgctgc tgaaggtgct gaagagaatc
     2701 agcgagaacc tgaagaagtt caaggagaag agcttcaagg agttcaaggg caagatcgag
     2761 agcaagatgc tgtacctggg cgaggaggtg aagctgctgg gcgagggcaa gatcaccggc
     2821 aagctggtgg gcctgagcga gaagggcggc gccctgatcc tgaccgagga gggcatcaag
     2881 gagatcctga gcggcgagtt cagcctgaga agaagctacc cttacgatgt accggattac
     2941 gcatgagtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3001 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3061 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3121 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3181 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3241 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     3301 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3361 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3421 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3481 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     3541 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3601 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3661 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     3721 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     3781 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3841 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     3901 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     3961 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     4021 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     4081 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     4141 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     4201 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4261 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     4321 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4381 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4441 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     4501 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     4561 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     4621 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     4681 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4741 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4801 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4861 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4921 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4981 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5041 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5101 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5161 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5221 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5281 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5341 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5401 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5461 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5521 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5581 agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5761 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6601 ccgcgcacat ttccccgaaa agtgccacct gcag
//
