LOCUS       sequence-467980-        8496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cleavable backbone for constitutive expression of TCR alpha and beta
            chains through lentiviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8496)
  AUTHORS   Afeyan AB, Wu CJ, Oliveira G
  TITLE     Rapid parallel reconstruction and specificity screening of hundreds 
            of T cell receptors.
  JOURNAL   Nat Protoc. 2024 Nov 8. doi: 10.1038/s41596-024-01061-4.
  PUBMED    39516267
REFERENCE   2  (bases 1 to 8496)
  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: "Nat Protoc.
            2024 Nov 8. doi: 10.1038/s41596-024-01061-4."
COMMENT     Sequence Label: PEW-pEF1A-inactEGFP
FEATURES             Location/Qualifiers
     source          1..8496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             633..813
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    860..985
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1478..1711
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1896..1940
                     /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    2207..2324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2372..3550
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2603..3541
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3498..3518
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3572..4285
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSA*PEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYI*ADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(3611..3632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3872..3891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4219..4240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4301..4889
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4354..4374)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4772..4783)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4976..5209
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5281..5402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5318..5337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5372..5391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5442..5577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5597..5616)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5598..5616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5626..5643)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5626..5642)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5635..5657)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5784..6239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5871..5890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6081..6102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6265..6369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6370..6438,6439..7230)
                     /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     complement(6588..6607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7401..7989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7890..7909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8143..8160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8277..8298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8313..8318,8319..8336,8337..8343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8351..8367
                     /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     8356..8378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8375..8391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8375..8391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8410..8430
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8412..8430
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctcg tttagtgaac cggggtctct ctggttagac cagatctgag
      661 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      721 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      781 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      841 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      901 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      961 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     1021 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     1081 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     1141 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1201 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1261 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1321 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     1381 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     1441 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     1501 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     1561 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     1621 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     1681 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     1741 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     1801 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     1861 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     1921 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     1981 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2041 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2101 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     2161 acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag
     2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttatcgat cacgagacta
     2341 gcctcgagaa gcttgatatc gaattccacg gggctccggt gcccgtcagt gggcagagcg
     2401 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     2461 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     2521 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     2581 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
     2641 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
     2701 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
     2761 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
     2821 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
     2881 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     2941 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     3001 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     3061 gtctcaagct ggccggcctg ctctggtgcc tggtctcgcg ccgccgtgta tcgccccgcc
     3121 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     3181 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     3241 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     3301 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     3361 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     3421 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     3481 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     3541 ggtgtcgtga ggggatccac cggtcgccac cagcaagggc gaggagctgt tcaccggggt
     3601 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3661 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3721 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3781 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcct aacccgaagg
     3841 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3901 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3961 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4021 tatctaagcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     4081 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4141 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4201 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     4261 cggcatggac gagctgtaca agtaaagcgg ccgcgtcgac aatcaacctc tggattacaa
     4321 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4381 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4441 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4501 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4561 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4621 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4681 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     4741 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4801 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4861 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacc tttaagacca
     4921 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     4981 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     5041 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5101 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5161 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     5221 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     5281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5401 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     5461 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5521 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5581 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     5641 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5701 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5761 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5821 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5881 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5941 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6001 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6061 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6121 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6181 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6241 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6301 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6361 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6421 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6481 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6541 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6601 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6661 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6721 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6781 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6841 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6901 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6961 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7021 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7081 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7141 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7201 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7261 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7321 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7381 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7441 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7501 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7561 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7621 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7681 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7741 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7801 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7861 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7921 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7981 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8041 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8101 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8161 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8221 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8281 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8341 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8401 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttggcca
     8461 ttgcatacgt tgtatccata tcataatatg tacatt
//
