LOCUS       sequence-441592-       10943 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 10943)
  AUTHORS   Uijttewaal ECH, Lee J, Sell AC, Botay N, Vainorius G, Novatchkova M,
            Baar J, Yang J, Potzler T, van der Leij S, Lowden C, Sinner J, 
            Elewaut A, Gavrilovic M, Obenauf A, Schramek D, Elling U
  TITLE     CRISPR-StAR enables high-resolution genetic screening in complex in 
            vivo models.
  JOURNAL   Nat Biotechnol. 2024 Dec 16. doi: 10.1038/s41587-024-02512-9.
  PUBMED    39681701
REFERENCE   2  (bases 1 to 10943)
  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: "Nat 
            Biotechnol. 2024 Dec 16. doi: 10.1038/s41587-024-02512-9."
COMMENT     Sequence Label: CRISPR-StAR4GN (Lenti)
FEATURES             Location/Qualifiers
     source          1..10943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(190..981,982..1050))
                     /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     813..832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1051..1155)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1230..1249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1425..1804
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1806..2004
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1959..1979
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2022..2202
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2249..2374
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2867..3100
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3285..3329
                     /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    3627..3744
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3872..4120
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3872..3892
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4043..4062
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(4164..4197)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4324..4357
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        4405..4904
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4440..4463)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4837..4856
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(4926..4928,4929..4931,4932..5642)
                     /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(4971..4992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5232..5251)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5579..5600
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5652..5708
                     /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             5703..6503
                     /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
                     IALATRDIAEELGGEWAGRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5763..5782)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6373..6392
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(6513..6546)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6626..6659
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    6818..7406
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6871..6891)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7289..7300)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7409..7425)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7410..7426)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7931..8111
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8137..8154)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8143..8367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8413..8841
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8500..8519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8710..8731
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8850..8870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8855..9184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9035..9170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9097..9116
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9232..9279
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9298..9672
                     /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    9802..9923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9839..9858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9893..9912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9972..9988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9972..9988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9985..10007)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9996..10012
                     /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(10020..10026,10027..10044,10045..10050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10065..10086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10203..10220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(10374..10943,1..19))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10454..10473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
       61 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
      121 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
      181 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     1201 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     1261 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     1321 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     1381 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgcg cgttgacatt gattattgac
     1441 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     1501 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1561 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     1621 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     1681 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1741 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1801 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     1861 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1921 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     1981 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     2041 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     2101 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     2161 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     2221 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     2281 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     2341 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     2401 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     2461 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     2521 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     2581 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     2641 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     2701 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     2761 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     2821 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2881 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2941 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     3001 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     3061 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     3121 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     3181 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     3241 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     3301 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     3361 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     3421 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     3481 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     3541 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     3601 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     3661 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     3721 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acaggctagc ctctaggcgc
     3781 cggaattgaa gatctccgcg gttaattaaa atgatacggc gaccaccgag atctgtcttg
     3841 atctaggagc tactcatcaa cggtcacatg tgagggccta tttcccatga ttccttcata
     3901 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
     3961 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     4021 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     4081 tttcttggct ttatatatct tgtggaaagg acgaaacacc cgagacgtct accgcacgtc
     4141 gtctcagttt aagagctatg ctgataactt cgtatagcat acattatacg aagttatttt
     4201 tttttttgcg gccgccaaga gagcagaaat agcaagtaac tcattaggta aataaggtag
     4261 tccgtttagt gccgtagcca attctcagac atttaatggc tctaaacgta acatttacaa
     4321 attataactt cgtataatgt gtactatacg aagttattaa ttaattctgg ttcgaatgta
     4381 gaattgtagc gatctaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     4441 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     4501 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     4561 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     4621 ggacgtgaca aatggaagta gcacctctca ctagtctcgt gcagatggac agcaccgctg
     4681 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     4741 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     4801 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     4861 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccagaatt
     4921 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4981 tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca
     5041 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     5101 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     5161 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5221 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5281 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5341 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5401 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5461 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5521 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaaa cgcgatcaca
     5581 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5641 agggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc
     5701 ccggccccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5761 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5821 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5881 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5941 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6001 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6061 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6121 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6181 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6241 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6301 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6361 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     6421 gccgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6481 gccttcttga cgagttcttc tgaacctgca ggataacttc gtatagcata cattatacga
     6541 agttatcagc atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac
     6601 cgagtcggtg cttttttgtt ttagcataac ttcgtataat gtgtactata cgaagttatc
     6661 tagaggtacc cgttaagatc ggaagagcac acgtctgaac tccagtcaca aaattatctc
     6721 gtatgccgtc ttatgcttgt taattaacaa ttgacgcgta attctaccgg ctagcgagag
     6781 cacacacgaa aggtaattcg atatcaagct tatcggtaat caacctctgg attacaaaat
     6841 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6901 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6961 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7021 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7081 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7141 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7201 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7261 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7321 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7381 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca
     7441 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     7501 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     7561 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     7621 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     7681 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     7741 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     7801 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     7861 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     7921 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     7981 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     8041 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     8101 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     8161 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     8221 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     8281 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     8341 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     8401 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     8461 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     8521 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     8581 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     8641 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     8701 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     8761 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     8821 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     8881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     8941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     9001 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     9061 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     9121 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     9181 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     9241 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     9301 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     9361 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     9421 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     9481 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     9541 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     9601 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     9661 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     9721 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     9781 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     9841 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     9901 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     9961 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    10021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    10081 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    10141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    10201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    10261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    10321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    10381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    10441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    10501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    10561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    10621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    10681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    10741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    10801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    10861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    10921 gtttgcaagc agcagattac gcg
//
