LOCUS       Exported                7726 bp ds-DNA     circular SYN 15-JUN-2022
DEFINITION  Expresses SARS-CoV-2 NSP2 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    NSP2-1x-BioID2 pBabe-puro (SARS-CoV-2)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7726)
  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 7726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3319..4014
                     /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             4015..4041
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4056..4076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4061..4390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4241..4376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4303..4322
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4400..4999
                     /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(4400..4419)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4896..4916
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5220..5689
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5819..5836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5990..6578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6070..6089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6738..7598)
                     /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     7361..7380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7599..7703)
                     /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 caccatggcc
     1381 tacacgcgat atgttgacaa caacttttgt ggacctgacg gctaccctct tgaatgtatt
     1441 aaggatcttc tggctcgcgc tggaaaagcc agttgtacac tgtctgagca gctcgacttt
     1501 atagatacaa agcggggggt ctattgttgc cgagagcatg aacatgagat cgcatggtat
     1561 acggagcgct ctgaaaaaag ctatgaattg cagacaccct ttgaaattaa gttggcgaaa
     1621 aagttcgata cattcaatgg tgagtgcccc aactttgtct ttccccttaa cagtatcatt
     1681 aaaacaatcc agccccgggt ggaaaagaag aaattggatg gattcatggg caggatcaga
     1741 agtgtttacc cagtggcatc ccctaatgaa tgcaatcaga tgtgcctgtc cacgctgatg
     1801 aaatgcgacc actgcggtga aacgtcttgg caaacggggg actttgtaaa agcaacgtgc
     1861 gagttttgcg gaactgagaa cttgacaaag gagggagcga ctacatgtgg atacctgcct
     1921 cagaacgcgg tagtgaaaat atactgtcct gcctgtcaca attctgaagt tgggcctgaa
     1981 catagcttgg ccgaatacca taacgaaagt ggattgaaga caattcttcg caaaggaggc
     2041 cggactatcg cgttcggggg ctgcgtgttt agctatgtag gatgtcataa taaatgcgca
     2101 tattgggttc cacgagctag cgcaaacata ggctgcaacc acacaggcgt agttggtgag
     2161 ggatcagaag gtcttaatga taacttgctc gaaatattgc aaaaggaaaa agtcaacata
     2221 aatattgtag gagattttaa attgaatgaa gagattgcga taatccttgc gtcattttcc
     2281 gcaagtacga gtgcgttcgt tgagactgta aaggggctcg actataaagc gtttaagcaa
     2341 atagtagaga gctgcggaaa ctttaaagtt acgaagggaa aagccaaaaa aggtgcatgg
     2401 aacattggcg agcagaagag tatactgtcc cccctgtacg ccttcgcgtc agaagcagct
     2461 cgagtagtta gaagtatatt ctcacgaacg ctcgagacgg cgcagaatag tgtgcgcgtg
     2521 ctccagaagg ctgctattac gatcctcgac ggcatatccc agtattctct tcgactgatt
     2581 gatgctatga tgttcacctc cgacttggcg acgaacaatc tcgtagttat ggcttatatc
     2641 accggcggcg ttgtacaatt gacgtctcag tggctgacga acatcttcgg aaccgtgtac
     2701 gagaagctca aaccagttct tgattggctt gaagaaaagt ttaaggaagg cgtcgagttt
     2761 ctccgggatg ggtgggaaat tgttaagttt atctccacct gcgcgtgtga aattgtggga
     2821 gggcaaattg tcacctgcgc caaggaaatt aaagagtcag ttcagacgtt ttttaaactt
     2881 gtgaacaaat tccttgctct ctgtgcggat agtataatca tcggtggggc gaagttgaaa
     2941 gcactcaatc ttggtgaaac tttcgtgacc cacagcaaag gcctgtatag gaagtgtgtt
     3001 aaaagcaggg aggagaccgg tcttcttatg ccgctgaagg cgcccaaaga gataattttt
     3061 cttgaggggg aaactcttcc gaccgaagtg ctcacggaag aagtagttct gaaaacaggg
     3121 gaccttcaac cactggagca accgacctct gaggcagttg aagcgccgct cgtcggaact
     3181 cctgtatgta ttaacggcct catgctgttg gaaattaagg atacggagaa gtactgtgca
     3241 ctggccccca acatgatggt cacgaacaac acctttaccc ttaagggcgg caccggtggt
     3301 ggaggcgggt ctgaattctt caagaacctg atctggctga aggaggtgga cagcacccag
     3361 gagagactga aggagtggaa cgtgagctac ggcaccgccc tggtggccga cagacagacc
     3421 aagggcagag gcggcctggg cagaaagtgg ctgagccagg agggcggcct gtacttcagc
     3481 ttcctgctga accccaagga gttcgagaac ctgctgcagc tgcccctggt gctgggcctg
     3541 agcgtgagcg aggccctgga ggagatcacc gagatcccct tcagcctgaa gtggcccaac
     3601 gacgtgtact tccaggagaa gaaggtgagc ggcgtgctgt gcgagctgag caaggacaag
     3661 ctgatcgtgg gcatcggcat caacgtgaac cagagagaga tccccgagga gatcaaggac
     3721 agagccacca ccctgtacga gatcaccggc aaggactggg acagaaagga ggtgctgctg
     3781 aaggtgctga agagaatcag cgagaacctg aagaagttca aggagaagag cttcaaggag
     3841 ttcaagggca agatcgagag caagatgctg tacctgggcg aggaggtgaa gctgctgggc
     3901 gagggcaaga tcaccggcaa gctggtgggc ctgagcgaga agggcggcgc cctgatcctg
     3961 accgaggagg gcatcaagga gatcctgagc ggcgagttca gcctgagaag aagctaccct
     4021 tacgatgtac cggattacgc atgagtcgac cctgtggaat gtgtgtcagt tagggtgtgg
     4081 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4141 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4201 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4261 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4321 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4381 cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4441 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4501 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4561 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4621 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4681 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     4741 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4801 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4861 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4921 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4981 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     5041 acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     5101 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     5161 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     5221 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     5281 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5341 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     5401 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     5461 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     5521 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     5581 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     5641 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     5701 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6541 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6601 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6661 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
     6721 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6781 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6841 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6901 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6961 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7021 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7081 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7141 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7201 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7261 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7321 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7381 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7441 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7501 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7561 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7621 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7681 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
