LOCUS       sequence-177650-        8178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Advanced lentiviral Vector for sgRNA (hU6 Promoter) delivery with 
            dTomato expression (EFS Promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8178)
  AUTHORS   Reimer J, Knoess S, Labuhn M, Charpentier EM, Gohring G, 
            Schlegelberger B, Klusmann JH, Heckl D
  TITLE     CRISPR-Cas9-induced t(11;19)/MLL-ENL translocations initiate 
            leukemia in human hematopoietic progenitor cells in vivo.
  JOURNAL   Haematologica. 2017 Jun 1. pii: haematol.2017.164046. doi: 
            10.3324/haematol.2017.164046.
  PUBMED    28572162
REFERENCE   2  (bases 1 to 8178)
  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: 
            "Haematologica. 2017 Jun 1. pii: haematol.2017.164046. doi: 
            10.3324/haematol.2017.164046."
COMMENT     Sequence Label: SGL40C.EFS.dTomato
FEATURES             Location/Qualifiers
     source          1..8178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    235..256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(271..276,277..294,295..301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    309..325
                     /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     314..336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     333..349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     333..349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     368..388
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        370..388
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        433..812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        869..1060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(892..912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1079..1305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1306..1486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1533..1658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2151..2384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2569..2613
                     /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    2911..3028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3079..3327
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3079..3099
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3250..3269
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3489..3700
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3720..3729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3726..4427
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3859..3877)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4127..4146
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4321..4344
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    4446..5034
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4499..4519)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4917..4928)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5106..5339
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5417..5538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5454..5473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5508..5527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5578..5713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5733..5752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5734..5752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5762..5779)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5762..5778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5771..5793)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5920..6375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6007..6026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6217..6238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6401..6505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6506..6574,6575..7366)
                     /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(6724..6743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7537..8125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8026..8045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
       61 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cggtaccgtc
      421 gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      601 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      781 acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct gaaataacct
      841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
      901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg
     1081 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     1141 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     1201 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     1261 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     1321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     2041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     2101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     2161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     2221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     2281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2761 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2821 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2881 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2941 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     3001 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     3061 ttggttaatt aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga
     3121 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     3181 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     3241 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     3301 tatatcttgt ggaaaggacg aaacaccgga gacgtatgtc gacaactcgt ctctgtttaa
     3361 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa
     3421 aagtggcacc gagtcggtgc ttttttgaat tctcctctag aactcgagtg gctccggtgc
     3481 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3541 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3601 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3661 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg
     3721 ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca
     3781 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3841 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     3901 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     3961 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     4021 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     4081 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     4141 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     4201 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     4261 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     4321 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     4381 agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaagtaa acgcgttaag
     4441 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4501 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4561 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4621 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4681 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4741 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4801 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4861 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4921 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4981 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac
     5041 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     5101 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     5161 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     5221 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     5281 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     5341 tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     5401 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5521 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     5581 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5641 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5701 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     5761 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     5821 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5881 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     5941 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6001 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6061 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6121 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6241 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6301 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6361 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6421 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6481 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6541 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6601 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6661 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6721 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6781 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6841 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6901 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6961 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7021 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7081 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7141 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7201 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7261 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7321 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7381 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7441 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7501 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7561 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7621 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7681 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7741 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7801 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7861 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7921 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7981 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8041 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8101 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8161 cttttgctgg ccttttgc
//
