LOCUS       sequence-352113-        8081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  construct expresses tet repressor (tetR) to reduce leakiness 
            together with the fluorochrome iRFP720.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   Liu WH, Mrozek-Gorska P, Wirth AK, Herold T, Schwarzkopf L, Pich D, 
            Volse K, Melo-Narvaez MC, Carlet M, Hammerschmidt W, Jeremias I
  TITLE     Inducible transgene expression in PDX models in vivo identifies KLF4
            as a therapeutic target for B-ALL.
  JOURNAL   Biomark Res. 2020 Sep 16;8:46. doi: 10.1186/s40364-020-00226-z. 
            eCollection 2020.
  PUBMED    32944247
REFERENCE   2  (bases 1 to 8081)
  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: "Biomark 
            Res."; date: "2020-09-16"; volume: "8:46. doi"; pages: " 
            10.1186/s40364-020-00226-z. eCollection 2020"
COMMENT     Sequence Label: pCDH-EF1a-tetR-T2A-iRFP
FEATURES             Location/Qualifiers
     source          1..8081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1966..2177
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2190..2458
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2504..3124
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2577..2596)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             3164..3217
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3224..4174
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     misc_feature    4181..4769
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4234..4254)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4652..4663)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4843..5076
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5148..5269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5185..5204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5239..5258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5288..5423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5456..5472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5456..5472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5469..5491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5480..5496
                     /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)."
     promoter        complement(join(5504..5510,5511..5528,5529..5534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5549..5570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5687..5704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5858..6446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6617..7408,7409..7477))
                     /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     7240..7259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7478..7582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7650..7668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7706..7728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7828..7847
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8041..8063
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8055..8072
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8056..8072
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     1921 tttatcgata ctagtggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     1981 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     2041 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2101 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2161 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     2221 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     2281 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     2341 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     2401 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     2461 agctgtgacc ggcgcctact ctagagctag cgaattcgcc accatgtcta gattagataa
     2521 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac
     2581 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa
     2641 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca
     2701 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt
     2761 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac
     2821 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc
     2881 actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt
     2941 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat
     3001 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt
     3061 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg
     3121 gtccgcgtac agcggatccc gggaattcgg atccgcggcc gctgagggca gaggaagtct
     3181 tctaacatgc ggtgacgtgg aggagaatcc cggccctatg catatggcgg aaggatccgt
     3241 cgccaggcag cctgacctct tgacctgcga cgatgagccg atccatatcc ccggtgccat
     3301 ccaaccgcat ggactgctgc tcgccctcgc cgccgacatg acgatcgttg ccggcagcga
     3361 caaccttccc gaactcaccg gactggcgat cggcgccctg atcggccgct ctgcggccga
     3421 tgtcttcgac tcggagacgc acaaccgtct gacgatcgcc ttggccgagc ccggggcggc
     3481 cgtcggagca ccgatcactg tcggcttcac gatgcgaaag gacgcaggct tcatcggctc
     3541 ctggcatcgc catgatcagc tcatcttcct cgagctcgag cctccccagc gggacgtcgc
     3601 cgagccgcag gcgttcttcc gccgcaccaa cagcgccatc cgccgcctgc aggccgccga
     3661 aaccttggaa agcgcctgcg ccgccgcggc gcaagaggtg cggaagatta ccggcttcga
     3721 tcgggtgatg atctatcgct tcgcctccga cttcagcggg tccgtgatcg cagaggatcg
     3781 gtgcgccgag gtcgagtcaa aactaggcct gcactatcct gcctcattca tcccggcgca
     3841 ggcccgtcgg ctctatacca tcaacccggt acggatcatt cccgatatca attatcggcc
     3901 ggtgccggtc accccagacc tcaatccggt caccgggcgg ccgattgatc ttagcttcgc
     3961 catcctgcgc agcgtctcgc ccaaccatct ggagttcatg cgcaacatag gcatgcacgg
     4021 cacgatgtcg atctcgattt tgcgcggcga gcgactgtgg ggattgatcg tttgccatca
     4081 ccgaacgccg tactacgtcg atctcgatgg ccgccaagcc tgcgagctag tcgcccaggt
     4141 tctggcctgg cagatcggcg tgatggaaga gtgagtcgac aatcaacctc tggattacaa
     4201 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4261 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4321 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4381 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4441 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4501 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4561 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     4621 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4681 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4741 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     4801 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     4861 cccaacgaag ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     4921 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     4981 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5041 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5101 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     5161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     5281 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5341 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5401 gtgaggaggc ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat
     5461 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5521 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5581 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5641 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5701 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5761 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5821 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5881 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6061 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6361 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6661 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6721 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6781 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6841 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6901 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6961 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7021 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7081 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7141 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7201 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7261 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7321 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7381 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7441 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7501 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7561 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7621 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     7681 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     7741 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7801 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7861 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     7921 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7981 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     8041 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
