LOCUS       sequence-364145-        6172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 ORF7a fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6172)
  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 6172)
  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: ORF7a-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    191..390
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             591..1007
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     937..959
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     975..991
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1414..2109
                     /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             2110..2136
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2151..2171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2156..2485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2336..2471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2495..3094
                     /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(2495..2514)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2991..3011
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3315..3784
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3914..3931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4833..5624,5625..5693))
                     /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     5456..5475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5694..5798)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(5829..6172,1..126)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
       61 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
      121 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
      181 accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga
      241 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc
      301 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
      361 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
      421 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
      481 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
      541 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
      601 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
      661 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
      721 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
      781 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
      841 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
      901 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
      961 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc cggatccgcc
     1021 accatgaaga tcatcctgtt cctggccctg atcaccctgg ctacctgcga gctgtaccac
     1081 taccaggaat gcgtgcgcgg taccaccgtg ctgctgaagg agccttgcag ctccggcacc
     1141 tacgaaggaa actccccctt ccaccctctg gccgacaaca agttcgctct gacctgcttc
     1201 tctactcagt tcgccttcgc ttgccctgat ggtgtgaagc acgtgtacca gctgcgcgcc
     1261 aggtctgtga gcccaaagct gttcatcaga caggaggaag tgcaggagct gtacagcccc
     1321 atcttcctga tcgtggccgc tatcgtgttc atcaccctgt gcttcaccct gaagcgtaag
     1381 accgaaaccg gtggtggagg cgggtctgaa ttcttcaaga acctgatctg gctgaaggag
     1441 gtggacagca cccaggagag actgaaggag tggaacgtga gctacggcac cgccctggtg
     1501 gccgacagac agaccaaggg cagaggcggc ctgggcagaa agtggctgag ccaggagggc
     1561 ggcctgtact tcagcttcct gctgaacccc aaggagttcg agaacctgct gcagctgccc
     1621 ctggtgctgg gcctgagcgt gagcgaggcc ctggaggaga tcaccgagat ccccttcagc
     1681 ctgaagtggc ccaacgacgt gtacttccag gagaagaagg tgagcggcgt gctgtgcgag
     1741 ctgagcaagg acaagctgat cgtgggcatc ggcatcaacg tgaaccagag agagatcccc
     1801 gaggagatca aggacagagc caccaccctg tacgagatca ccggcaagga ctgggacaga
     1861 aaggaggtgc tgctgaaggt gctgaagaga atcagcgaga acctgaagaa gttcaaggag
     1921 aagagcttca aggagttcaa gggcaagatc gagagcaaga tgctgtacct gggcgaggag
     1981 gtgaagctgc tgggcgaggg caagatcacc ggcaagctgg tgggcctgag cgagaagggc
     2041 ggcgccctga tcctgaccga ggagggcatc aaggagatcc tgagcggcga gttcagcctg
     2101 agaagaagct acccttacga tgtaccggat tacgcatgag tcgaccctgt ggaatgtgtg
     2161 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2221 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2281 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2341 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2401 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2461 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     2521 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     2581 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     2641 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     2701 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     2761 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     2821 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     2881 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     2941 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3001 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     3061 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     3121 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     3181 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3241 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     3301 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3361 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     3421 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     3481 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     3541 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     3601 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     3661 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     3721 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     3781 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     3841 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     4801 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4861 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4921 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4981 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5041 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5101 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5161 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5221 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5281 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5341 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5401 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5461 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5521 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5581 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5641 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5701 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5761 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5821 cctgcagcct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     5881 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     5941 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     6001 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     6061 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     6121 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga tt
//
