LOCUS       sequence-441457-       10694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10694)
  AUTHORS   Michlits G, Hubmann M, Wu SH, Vainorius G, Budusan E, Zhuk S, 
            Burkard TR, Novatchkova M, Aichinger M, Lu Y, Reece-Hoyes J, Nitsch 
            R, Schramek D, Hoepfner D, Elling U
  TITLE     CRISPR-UMI: single-cell lineage tracing of pooled CRISPR-Cas9 
            screens.
  JOURNAL   Nat Methods. 2017 Dec;14(12):1191-1197. doi: 10.1038/nmeth.4466. 
            Epub 2017 Oct 16.
  PUBMED    29039415
REFERENCE   2  (bases 1 to 10694)
  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"; doi: 
            "10.1038/nmeth.4466"; journalName: "Nat Methods"; date: "2017-12"; 
            volume: "14"; issue: "12"; pages: "1191-1197"
COMMENT     Sequence Label: pLenti-UMI
FEATURES             Location/Qualifiers
     source          1..10694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             132..312
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(338..355)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    344..568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      614..1042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(701..720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     911..932
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1051..1071)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1056..1385
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1236..1371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1298..1317
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1433..1480
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1499..1873
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2003..2124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2040..2059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2094..2113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2173..2189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2173..2189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2186..2208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2197..2213
                     /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(2221..2227,2228..2245,2246..2251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2266..2287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2404..2421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2575..3163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2655..2674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3334..4125,4126..4194))
                     /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     3957..3976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4195..4299)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4374..4393)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4569..4948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4950..5148
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5103..5123
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5166..5346
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5393..5518
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6011..6244
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6429..6473
                     /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    6771..6888
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7016..7264
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7016..7036
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7187..7206
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        7592..8091
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7627..7650)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8024..8043
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(8113..8115,8116..8118,8119..8829)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8158..8179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8419..8438)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8766..8787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8839..8895
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8890..9690
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8950..8969)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9560..9579
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    9713..10301
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9766..9786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10184..10195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10304..10320)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10305..10321)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
       61 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
      121 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
      181 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
      241 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
      301 aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
      361 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
      421 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
      481 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
      541 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
      601 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      661 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      721 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      781 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      841 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      901 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      961 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1021 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     1081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1141 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1201 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     1261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1321 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1381 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     1441 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     1501 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
     1561 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
     1621 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
     1681 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
     1741 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
     1801 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
     1861 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     1921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1981 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     2041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2101 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     2161 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2221 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2281 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2401 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3001 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3061 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3541 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3601 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3661 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3721 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3781 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3841 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3901 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3961 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4021 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4081 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4141 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4201 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4261 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4321 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     4381 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     4441 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     4501 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     4561 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     4621 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     4681 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     4741 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     4801 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     4861 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     4921 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     4981 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     5041 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     5101 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct
     5161 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5221 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5281 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5341 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     5401 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     5461 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     5521 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     5581 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     5641 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     5701 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     5761 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     5821 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     5881 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     5941 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     6001 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     6061 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     6121 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     6181 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     6241 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     6301 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     6361 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     6421 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     6481 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     6541 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     6601 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     6661 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     6721 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     6781 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     6841 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     6901 agggacaggc tagcctctag gcgccggaat tgaagatctc cgcggttaat taaaatgata
     6961 cggcgaccac cgagatctgt cttgatctag gagctactca tcaacggtca catgtgaggg
     7021 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     7081 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     7141 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
     7201 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
     7261 cacccgagac gtctaccgca cgtcgtctca gtttaagagc tatgctggaa acagcatagc
     7321 aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     7381 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     7441 caattctgca gacaaatggc tctagaggta cccgttaaga tcggaagagc acacgtctga
     7501 actccagtca caaaattatc tcgtatgccg tcttatgctt gttaattaac aattgacgcg
     7561 taattctacc ggctagcgat ctaattctac cgggtagggg aggcgctttt cccaaggcag
     7621 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     7681 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     7741 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     7801 tcgtgcagga cgtgacaaat ggaagtagca cctctcacta gtctcgtgca gatggacagc
     7861 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     7921 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     7981 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     8041 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     8101 cagaattcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     8161 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc
     8221 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     8281 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     8341 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     8401 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     8461 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     8521 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     8581 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc
     8641 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     8701 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaaacgc
     8761 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     8821 ctgtacaagg gcagcggcgc caccaacttc agcctgctga agcaggccgg cgacgtggag
     8881 gagaaccccg gccccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     8941 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     9001 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     9061 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     9121 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     9181 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     9241 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     9301 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     9361 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     9421 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     9481 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     9541 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     9601 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     9661 ttctatcgcc ttcttgacga gttcttctga attcgatatc aagcttatcg gtaatcaacc
     9721 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     9781 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     9841 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     9901 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     9961 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
    10021 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
    10081 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
    10141 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
    10201 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
    10261 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
    10321 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg
    10381 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc
    10441 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
    10501 gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta
    10561 ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata
    10621 tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga
    10681 agccaatgaa ggag
//
