LOCUS       sequence-364142-        6570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 Membrane protein fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6570)
  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 6570)
  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: M-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2068..2763
                     /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             2764..2790
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2805..2825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2810..3139
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2990..3125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3149..3748
                     /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(3149..3168)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3645..3665
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4645..4662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4816..5404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4896..4915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5582..6373,6374..6442))
                     /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     6205..6224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6443..6547)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccgc caccatggct
     1381 gactctaacg gtaccatcac cgtggaggaa ctgaagaagc tgctggagca gtggaacctg
     1441 gtcatcggct tcctgttcct gacctggatc tgcctgctgc agttcgccta cgctaaccgc
     1501 aacaggttcc tgtacatcat caagctgatc ttcctgtggc tgctgtggcc tgtgaccctg
     1561 gcttgcttcg tgctggctgc cgtgtaccgc atcaactgga tcaccggcgg aatcgccatc
     1621 gctatggcct gcctggtggg cctgatgtgg ctgtcttact tcatcgctag cttcaggctg
     1681 ttcgccagaa cccgttccat gtggtctttc aaccccgaga ccaacatcct gctgaacgtg
     1741 cctctgcacg gaaccatcct gaccagacca ctgctggaga gcgaactggt catcggcgct
     1801 gtgatcctga gaggacacct gcgtatcgcc ggacaccacc tgggtcgttg cgacatcaag
     1861 gacctgccca aggaaatcac cgtggctacc agccgcaccc tgtcctacta caagctggga
     1921 gcttctcaga gagtggctgg tgactctggt ttcgctgctt actctcgcta caggatcggt
     1981 aactacaagc tgaacaccga ccacagctcc tctagcgaca acatcgccct gctggtgcag
     2041 accggtggtg gaggcgggtc tgaattcttc aagaacctga tctggctgaa ggaggtggac
     2101 agcacccagg agagactgaa ggagtggaac gtgagctacg gcaccgccct ggtggccgac
     2161 agacagacca agggcagagg cggcctgggc agaaagtggc tgagccagga gggcggcctg
     2221 tacttcagct tcctgctgaa ccccaaggag ttcgagaacc tgctgcagct gcccctggtg
     2281 ctgggcctga gcgtgagcga ggccctggag gagatcaccg agatcccctt cagcctgaag
     2341 tggcccaacg acgtgtactt ccaggagaag aaggtgagcg gcgtgctgtg cgagctgagc
     2401 aaggacaagc tgatcgtggg catcggcatc aacgtgaacc agagagagat ccccgaggag
     2461 atcaaggaca gagccaccac cctgtacgag atcaccggca aggactggga cagaaaggag
     2521 gtgctgctga aggtgctgaa gagaatcagc gagaacctga agaagttcaa ggagaagagc
     2581 ttcaaggagt tcaagggcaa gatcgagagc aagatgctgt acctgggcga ggaggtgaag
     2641 ctgctgggcg agggcaagat caccggcaag ctggtgggcc tgagcgagaa gggcggcgcc
     2701 ctgatcctga ccgaggaggg catcaaggag atcctgagcg gcgagttcag cctgagaaga
     2761 agctaccctt acgatgtacc ggattacgca tgagtcgacc ctgtggaatg tgtgtcagtt
     2821 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3001 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3061 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3121 aggcctaggc ttttgcaaaa agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     3181 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     3241 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     3301 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     3361 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     3421 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     3481 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     3541 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     3601 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     3661 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     3721 gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga
     3781 aaggagcgca cgaccccatg catcgataaa ataaaagatt ttatttagtc tccagaaaaa
     3841 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     3901 caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga
     3961 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     4021 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     4081 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     4141 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4201 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4261 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     4321 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4381 ctgttccttg ggagggtctc ctctgagtga ttgactmcmm kwmascsksk kkcwktyrca
     4441 tscgrcttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4501 agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     4561 ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4621 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4681 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4741 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4801 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4861 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4921 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4981 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5041 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5101 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5161 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5221 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5281 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5341 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5401 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5461 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc
     5521 gcaaatcaat ctaaagtatm wawkmrwwmw mwwgkwtwta tgagtaaact tggtctgaca
     5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6541 ttccgcgcac atttccccga aaagtgccac
//
