LOCUS       sequence-237867-        8190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retrovirus for delivery of one sgRNA (self-targeting) - 
            GFP-targeting control plasmid contains GFP instead of PuroR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8190)
  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 8190)
  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(gfp)U6sgRNA5(BbsI)-PGKpuroBFP
FEATURES             Location/Qualifiers
     source          1..8190
                     /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        5535..6034
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5570..5593)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5967..5986
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      6058..6067
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6064..6765
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             6784..6837
                     /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(6844..6846,6847..6849,6850..7563)
                     /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(6844..6846,6847..6849,6850..7560)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6889..6910)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7150..7169)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7497..7518
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7623..7643)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8041..8052)
                     /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 aacaccgaag ttcgagggcg acacccgttt aagagctatg ctggaaacag catagcaagt
     5461 ttaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttg
     5521 gatccaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     5581 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     5641 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     5701 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     5761 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     5821 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     5881 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     5941 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     6001 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc atccatctag atctcgagcc
     6061 accatgggca gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     6121 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     6181 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     6241 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     6301 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaggac
     6361 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     6421 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     6481 ggctgggagg ccttcaccga gaccctgtac cccgctgacg gcggcctgga aggcagaaac
     6541 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     6601 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     6661 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     6721 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaatggagg gggcagcggc
     6781 ggggaaggaa ggggctccct gctcacctgc ggcgacgtgg aagagaaccc cggacctgaa
     6841 ttcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     6901 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     6961 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     7021 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     7081 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     7141 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     7201 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     7261 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     7321 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     7381 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     7441 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     7501 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     7561 taagcggccg ctcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     7621 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     7681 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     7741 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     7801 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     7861 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     7921 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     7981 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     8041 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     8101 ccgcgtctac gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcag
     8161 gtacctttaa gaccaatgac ttacaaggat
//
