LOCUS       sequence-237863-        8188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retrovirus for testing CRISPR KO activity (empty) - non-targeting 
            control plasmid contains GFP instead of PuroR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8188)
  AUTHORS   Henriksson J, Chen X, Gomes T, Ullah U, Meyer KB, Miragaia R, Duddy 
            G, Pramanik J, Yusa K, Lahesmaa R, Teichmann SA
  TITLE     Genome-wide CRISPR Screens in T Helper Cells Reveal Pervasive 
            Crosstalk between Activation and Differentiation.
  JOURNAL   Cell. 2019 Jan 7. pii: S0092-8674(18)31569-1. doi: 
            10.1016/j.cell.2018.11.044.
  PUBMED    30639098
REFERENCE   2  (bases 1 to 8188)
  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: "Cell. 2019 
            Jan 7. pii: S0092-8674(18)31569-1. doi: 10.1016/j.cell.2018.11.044."
COMMENT     Sequence Label: pMSCV(W-)U6sgRNA5(BbsI)-PGKpuroBFP
FEATURES             Location/Qualifiers
     source          1..8188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             45..559
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    721..737
                     /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(745..751,752..769,770..775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    790..811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(928..945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1099..1687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1179..1198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1858..2649,2650..2718))
                     /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     2481..2500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2719..2823)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2891..2909
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2947..2969)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3069..3088
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3282..3304
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3432..3451
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             3651..4167
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    4231..4572
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             4639..5055
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4985..5007
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5023..5039
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        5158..5406
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5158..5178
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5329..5348
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        5533..6032
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5568..5591)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5965..5984
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      6056..6065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6062..6763
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             6782..6835
                     /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             join(6842..6844,6845..6847,6848..7561)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6842..6844,6845..6847,6848..7558)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6887..6908)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7148..7167)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7495..7516
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7621..7641)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8039..8050)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
       61 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
      121 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
      181 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
      241 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
      301 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
      361 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
      421 agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
      481 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
      541 gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg
      601 taggagtcga atattaagta atcctgactc aattagccac tgttttgaat ccacatactc
      661 caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa
      721 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      781 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      841 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1501 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2821 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2881 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2941 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3001 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     3061 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     3121 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     3181 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3241 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3301 aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     3361 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     3421 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     3481 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata
     3541 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     3601 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     3661 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     3721 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     3781 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3841 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3901 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     3961 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4021 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct
     4081 tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt
     4141 gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc
     4201 cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc
     4261 tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta
     4321 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc
     4381 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga
     4441 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac
     4501 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg
     4561 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
     4621 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
     4681 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
     4741 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
     4801 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
     4861 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
     4921 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
     4981 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
     5041 ccttctctag gcgccggaat tcagatctta cgtagctagc ggatcccaat tgcacaggga
     5101 cagcagagat ccagtttggt tagtaccggg ccctacgcgt ccaaggtcgg gcaggaagag
     5161 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     5221 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     5281 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     5341 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     5401 aacaccgggt cttcgagaag acacgtttaa gagctatgct ggaaacagca tagcaagttt
     5461 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgga
     5521 tccaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     5581 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     5641 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     5701 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     5761 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     5821 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     5881 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     5941 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     6001 tgttctcctc ttcctcatct ccgggccttt cgacctgcat ccatctagat ctcgagccac
     6061 catgggcagc gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt
     6121 ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
     6181 gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     6241 tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt
     6301 cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat acgaggacgg
     6361 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     6421 caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg
     6481 ctgggaggcc ttcaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagaaacga
     6541 catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga ccacatatag
     6601 atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact
     6661 ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc
     6721 cagatactgc gacctcccta gcaaactggg gcacaagctt aatggagggg gcagcggcgg
     6781 ggaaggaagg ggctccctgc tcacctgcgg cgacgtggaa gagaaccccg gacctgaatt
     6841 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     6901 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     6961 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     7021 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     7081 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     7141 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     7201 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     7261 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     7321 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     7381 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     7441 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     7501 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     7561 agcggccgct caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     7621 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     7681 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     7741 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     7801 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     7861 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     7921 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     7981 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     8041 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     8101 gcgtctacgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcaggt
     8161 acctttaaga ccaatgactt acaaggat
//
