LOCUS       Exported                6682 bp ds-DNA     circular SYN 15-JUN-2022
DEFINITION  Expresses SARS-CoV-2 NSP6 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    NSP6-1x-BioID2 pBabe-puro (SARS-CoV-2)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6682)
  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 6682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6682
                     /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"
     CDS             2275..2970
                     /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             2971..2997
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3012..3032)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3017..3346
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3197..3332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3259..3278
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3356..3955
                     /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(3356..3375)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3852..3872
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4176..4645
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4775..4792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4946..5534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5026..5045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5694..6554)
                     /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     6317..6336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6555..6659)
                     /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 caccatgagc
     1381 gccgtgaaga gaaccatcaa gggaacccac cactggctgc tgctgaccat cctgacctcc
     1441 ctgctggtgc tggtgcagtc tacccagtgg agcctgttct tcttcctgta cgagaacgcc
     1501 ttcctgccct tcgctatggg tatcatcgcc atgtccgcct tcgctatgat gttcgtgaag
     1561 cacaagcacg ctttcctgtg cctgttcctg ctgccatccc tggccaccgt ggcttacttc
     1621 aacatggtgt acatgcccgc ctcttgggtc atgcgcatca tgacctggct ggacatggtg
     1681 gacaccagcc tgtccggctt caagctgaag gactgcgtga tgtacgcctc tgctgtggtg
     1741 ctgctgatcc tgatgaccgc tagaaccgtg tacgatgatg gtgctagaag ggtgtggacc
     1801 ctgatgaacg tgctgaccct ggtgtacaag gtgtactacg gcaacgccct ggaccaggct
     1861 atcagcatgt gggctctgat catctctgtg accagcaact actccggagt ggtgaccacc
     1921 gtgatgttcc tggccagagg tatcgtgttc atgtgcgtgg agtactgccc tatcttcttc
     1981 atcaccggca acaccctgca gtgcatcatg ctggtgtact gcttcctggg ttacttctgc
     2041 acctgctatt tcggcctgtt ctgcctgctg aacagatact tccgtctgac cctgggcgtg
     2101 tacgactacc tggtgtccac ccaggaattc cgttacatga actctcaggg actgctgccc
     2161 cctaagaaca gcatcgacgc tttcaagctg aacatcaagc tgctgggagt gggcggaaag
     2221 ccatgcatca aggtggccac cgtgcagacc ggtggtggag gcgggtctga attcttcaag
     2281 aacctgatct ggctgaagga ggtggacagc acccaggaga gactgaagga gtggaacgtg
     2341 agctacggca ccgccctggt ggccgacaga cagaccaagg gcagaggcgg cctgggcaga
     2401 aagtggctga gccaggaggg cggcctgtac ttcagcttcc tgctgaaccc caaggagttc
     2461 gagaacctgc tgcagctgcc cctggtgctg ggcctgagcg tgagcgaggc cctggaggag
     2521 atcaccgaga tccccttcag cctgaagtgg cccaacgacg tgtacttcca ggagaagaag
     2581 gtgagcggcg tgctgtgcga gctgagcaag gacaagctga tcgtgggcat cggcatcaac
     2641 gtgaaccaga gagagatccc cgaggagatc aaggacagag ccaccaccct gtacgagatc
     2701 accggcaagg actgggacag aaaggaggtg ctgctgaagg tgctgaagag aatcagcgag
     2761 aacctgaaga agttcaagga gaagagcttc aaggagttca agggcaagat cgagagcaag
     2821 atgctgtacc tgggcgagga ggtgaagctg ctgggcgagg gcaagatcac cggcaagctg
     2881 gtgggcctga gcgagaaggg cggcgccctg atcctgaccg aggagggcat caaggagatc
     2941 ctgagcggcg agttcagcct gagaagaagc tacccttacg atgtaccgga ttacgcatga
     3001 gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3181 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3241 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3301 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaccatgac
     3361 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3421 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3481 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3541 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3601 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3661 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3721 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3781 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3841 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3901 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc
     3961 gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgataaaata
     4021 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     4081 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     4141 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     4201 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     4261 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4321 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     4381 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     4441 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4501 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     4561 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     4621 actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt
     4681 tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga
     4741 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4801 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4861 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4921 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5461 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5521 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5581 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5641 ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6661 acatttcccc gaaaagtgcc ac
//
