LOCUS       sequence-255499-        8375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BirA* expressed as a protein fusion with MS2 coat protein (MCP) 
            monomer and EGFP; 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 8375)
  TITLE     Jansen lab plasmids
REFERENCE   2  (bases 1 to 8375)
  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 - mcp - egfp - BirA*
FEATURES             Location/Qualifiers
     source          1..8375
                     /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             join(3063..3065,3066..3068,3069..3779)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3063..3065,3066..3068,3069..3779)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3108..3129)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3369..3388)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3716..3737
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(3789..4136,4137..4139,4140..4748)
                     /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    4765..5353
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4818..4838)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5236..5247)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5428..5661
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5780..5798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5866..5970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5971..6039,6040..6831)
                     /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(6189..6208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7002..7590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7491..7510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7744..7761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8001..8022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8037..8042,8043..8060,8061..8067)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8075..8091
                     /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     8080..8102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8099..8115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8099..8115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8165..8184
                     /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 cgccatatga
     2701 tggcttctaa ctttactcag ttcgttctcg tcgacaatgg cggaactggc gacgtgactg
     2761 tcgccccaag caacttcgct aacgggatcg ctgaatggat cagctctaac tcgcgttcac
     2821 aggcttacaa agtaacctgt agcgttcgtc agagctctgc gcagaatcgc aaatacacca
     2881 tcaaagtcga ggtgcctaaa ggcgcctggc gttcgtactt aaatatggaa ctaaccattc
     2941 caattttcgc cacgaattcc gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa
     3001 aagatggaaa cccgattccc tcagcaatcg cagcaaactc cggcatctac gcggattcta
     3061 gaatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3121 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3181 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3241 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3301 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3361 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3421 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3481 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3541 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3601 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3661 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3721 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc
     3781 tcgaggacaa ggacaacacc gtgcccctga agctgatcgc cctgctggcc aacggcgagt
     3841 tccactctgg cgagcagctg ggagagaccc tgggaatgag cagagccgcc atcaacaagc
     3901 acatccagac actgagagac tggggagtgg acgtgttcac cgtgcctggc aagggctaca
     3961 gcctgcctga gcctatccag ctgctgaacg ccaagcagat cctgggacag ctggatggcg
     4021 gaagcgtggc cgtgctgcct gtgatcgact ccaccaatca gtacctgctg gacagaatcg
     4081 gagagctgaa gtccggcgac gcctgcatcg ccgagtacca gcaggctggc agaggaggca
     4141 gaggacggaa gtggttcagc ccattcggag ccaacctgta cctgtccatg ttctggagac
     4201 tggagcaggg acctgctgct gccatcggac tgagtctggt gatcggaatc gtgatggccg
     4261 aggtgctgag aaagctggga gccgacaagg tgagagtgaa gtggcctaat gacctgtacc
     4321 tccaggaccg caagctggct ggcatcctgg tggagctgac aggcaagaca ggcgatgccg
     4381 ctcagatcgt gatcggagcc ggaatcaaca tggccatgag aagagtggag gagagcgtgg
     4441 tgaaccaggg ctggatcacc ctgcaggagg ctggcatcaa cctggaccgg aacaccctgg
     4501 ccgccatgct gatcagagag ctgagagccg ctctggagct gttcgagcag gagggactgg
     4561 ctccttacct gagcagatgg gagaagctgg acaacttcat caacagacct gtgaagctga
     4621 tcatcggcga caaggaaatc ttcggcatct ccagaggaat cgacaagcag ggagctctgc
     4681 tgctggagca ggacggaatc atcaagccct ggatgggcgg agaaatctcc ctgagaagcg
     4741 cagagaagta aatcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg
     4801 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4861 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4921 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4981 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5041 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5101 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5161 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5221 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5281 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5341 gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta
     5401 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     5461 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5521 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5581 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5641 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5701 ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat
     5761 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5821 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5941 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6001 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6061 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6121 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6181 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     6241 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6301 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6361 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6421 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6481 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6541 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6601 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6661 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6721 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6781 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6841 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6901 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6961 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7021 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7081 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7141 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7201 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7261 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7321 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7381 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7441 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7501 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7561 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7621 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7681 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7741 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7801 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg
     7861 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7921 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg
     7981 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8041 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8101 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct
     8161 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     8221 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     8281 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     8341 tgtatttgtt aaatatgtac tacaaactta gtagt
//
