LOCUS       sequence-255502-        8732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BirA* expressed as a protein fusion with MS2 coat protein (MCP) 
            dimer and mCherry; to visualize any MS2 tagged mRNAs and to identify
            the RNA proximal proteome by their biotinylation. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8732)
  TITLE     Jansen lab plasmids
REFERENCE   2  (bases 1 to 8732)
  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"
COMMENT     Sequence Label: phage - ubc - nls - 2xmcp - mCherry - BirA*
FEATURES             Location/Qualifiers
     source          1..8732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2218..2616
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             2631..2651
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2655..2681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2655..2681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2682..2693
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3429..4136
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3574..3592)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3842..3861
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(4146..4493,4494..4496,4497..5105)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     misc_feature    5122..5710
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5175..5195)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5593..5604)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5785..6018
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6137..6155)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6223..6327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6328..6396,6397..7188)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6546..6565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7359..7947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7848..7867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8101..8118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8358..8379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8394..8399,8400..8417,8418..8424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8432..8448
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8437..8459
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8456..8472
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8456..8472
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8522..8541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
     2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2461 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgcaatgggc ccaaaaaaga
     2641 aaagaaaagt tggctacccc tacgacgtgc ccgactacgc catcgaaggc cgccatatgc
     2701 tagccgttaa aatggcttct aactttactc agttcgttct cgtcgacaat ggcggaactg
     2761 gcgacgtgac tgtcgcccca agcaacttcg ctaacgggat cgctgaatgg atcagctcta
     2821 actcgcgttc acaggcttac aaagtaacct gtagcgttcg tcagagctct gcgcagaatc
     2881 gcaaatacac catcaaagtc gaggtgccta aaggcgcctg gcgttcgtac ttaaatatgg
     2941 aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt aaggcaatgc
     3001 aaggtctcct aaaagatgga aacccgattc cctcagcaat cgcagcaaac tccggcatct
     3061 acgccatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga actggcgacg
     3121 tgactgtcgc cccaagcaac ttcgctaacg ggatcgctga atggatcagc tctaactcgc
     3181 gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag aatcgcaaat
     3241 acaccatcaa agtcgaggtg cctaaaggcg cctggcgttc gtacttaaat atggaactaa
     3301 ccattccaat tttcgccacg aattccgact gcgagcttat tgttaaggca atgcaaggtc
     3361 tcctaaaaga tggaaacccg attccctcag caatcgcagc aaactccggc atctacgcgg
     3421 attctagaat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     3481 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     3541 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     3601 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     3661 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     3721 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     3781 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     3841 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     3901 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     3961 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     4021 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     4081 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagctcg
     4141 aggacaagga caacaccgtg cccctgaagc tgatcgccct gctggccaac ggcgagttcc
     4201 actctggcga gcagctggga gagaccctgg gaatgagcag agccgccatc aacaagcaca
     4261 tccagacact gagagactgg ggagtggacg tgttcaccgt gcctggcaag ggctacagcc
     4321 tgcctgagcc tatccagctg ctgaacgcca agcagatcct gggacagctg gatggcggaa
     4381 gcgtggccgt gctgcctgtg atcgactcca ccaatcagta cctgctggac agaatcggag
     4441 agctgaagtc cggcgacgcc tgcatcgccg agtaccagca ggctggcaga ggaggcagag
     4501 gacggaagtg gttcagccca ttcggagcca acctgtacct gtccatgttc tggagactgg
     4561 agcagggacc tgctgctgcc atcggactga gtctggtgat cggaatcgtg atggccgagg
     4621 tgctgagaaa gctgggagcc gacaaggtga gagtgaagtg gcctaatgac ctgtacctcc
     4681 aggaccgcaa gctggctggc atcctggtgg agctgacagg caagacaggc gatgccgctc
     4741 agatcgtgat cggagccgga atcaacatgg ccatgagaag agtggaggag agcgtggtga
     4801 accagggctg gatcaccctg caggaggctg gcatcaacct ggaccggaac accctggccg
     4861 ccatgctgat cagagagctg agagccgctc tggagctgtt cgagcaggag ggactggctc
     4921 cttacctgag cagatgggag aagctggaca acttcatcaa cagacctgtg aagctgatca
     4981 tcggcgacaa ggaaatcttc ggcatctcca gaggaatcga caagcaggga gctctgctgc
     5041 tggagcagga cggaatcatc aagccctgga tgggcggaga aatctccctg agaagcgcag
     5101 agaagtaaat cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact
     5161 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5221 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5281 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5341 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5401 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5461 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5521 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5581 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5641 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5701 gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5761 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     5821 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5881 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5941 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6001 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     6061 caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat
     6121 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6181 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6241 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6301 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6361 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6421 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6481 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6541 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     6601 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6661 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6721 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6781 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6841 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6901 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6961 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7021 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7081 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7141 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7201 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7261 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7321 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7381 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7441 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7501 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7561 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7621 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7681 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7741 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7801 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7861 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7921 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7981 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8041 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8101 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8161 cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag
     8221 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     8281 gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc
     8341 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8401 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8461 aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt
     8521 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
     8581 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
     8641 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
     8701 atttgttaaa tatgtactac aaacttagta gt
//
