LOCUS       sequence-298974-        8319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of crRNA with GFP and a neomycin resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8319)
  AUTHORS   Gier RA, Budinich KA, Evitt NH, Cao Z, Freilich ES, Chen Q, Qi J, 
            Lan Y, Kohli RM, Shi J
  TITLE     High-performance CRISPR-Cas12a genome editing for combinatorial 
            genetic screening.
  JOURNAL   Nat Commun. 2020 Jul 13;11(1):3455. doi: 10.1038/s41467-020-17209-1.
  PUBMED    32661245
REFERENCE   2  (bases 1 to 8319)
  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 
            Commun."; date: "2020-07-13"; volume: "11(1):3455. doi"; pages: " 
            10.1038/s41467-020-17209-1"
COMMENT     Sequence Label: EFS-EGFP-P2A-Neo-WPRE-U6-AsCas12aDR-AsCas12aDR...
FEATURES             Location/Qualifiers
     source          1..8319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    64..181
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        265..476
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      496..505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             505..528
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(529..531,532..534,535..1245)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(529..531,532..534,535..1245)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(574..595)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(835..854)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1182..1203
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1255..1311
                     /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             1312..2106
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1366..1385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1976..1995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    2122..2710
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2175..2195)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2593..2604)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2866..3114
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2866..2886
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3037..3056
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     LTR             3233..3413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    3491..3612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3528..3547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3582..3601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3652..3787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3808..3826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3836..3853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3836..3852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3845..3867)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3994..4449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4081..4100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4291..4312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4475..4579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4580..4648,4649..5440)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4798..4817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5611..6199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6100..6119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6353..6370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6487..6508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6523..6528,6529..6546,6547..6553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6561..6577
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6566..6588
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6585..6601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6585..6601
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6620..6640
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6622..6640
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6668..6894
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6895..7075
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7122..7247
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7740..7973
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8158..8202
                     /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"
ORIGIN
        1 aggcagggat attcaccatt atcgtttcag aatcttgaga caaatggcag tattcatcca
       61 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
      121 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
      181 tcgggtttat tacagggaca gcagagatcc actttggcgc cggctcgagg gggcccgggt
      241 cgagtggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      301 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
      361 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      421 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtgt
      481 cgtgacgcgg gatccgccac catggattac aaagacgatg acgataagat ggtgagcaag
      541 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      601 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      661 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      721 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      781 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      841 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      901 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      961 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1021 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1081 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1141 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1201 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcag cggcgccacc
     1261 aacttcagcc tgctgaagca ggccggcgac gtggaggaga accccggccc catgcttgaa
     1321 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     1381 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     1441 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     1501 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     1561 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     1621 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     1681 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     1741 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     1801 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     1861 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     1921 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     1981 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     2041 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     2101 ttctgaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     2161 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2221 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2281 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2341 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2401 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2461 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     2521 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2581 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2641 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2701 gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact
     2761 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatttca
     2821 ccattatcgt ttcagaccca cctcccaacc ccgagtggac ccagagaggg cctatttccc
     2881 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tagaattaat
     2941 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     3001 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     3061 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgaattt
     3121 ctactcttgt agatcgagac ggttgtattt gtacgtctca aatttctact cttgtagatt
     3181 ttttttaagc ttggcgtaac tagatcttga gacactgctt tttgcttgta ctgggtctct
     3241 cttgttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     3301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     3361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta
     3421 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     3481 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3601 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     3661 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3721 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3781 ggaggcctag ggacgtaccc aattcgtcct atagtgagtc gtattacgcg cgctcactgg
     3841 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3901 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3961 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     4021 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4081 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4141 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4201 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4261 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4321 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4381 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4441 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4501 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4561 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4621 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4681 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4741 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4801 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4861 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4921 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4981 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5041 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5101 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5161 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5221 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5281 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5341 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5401 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5461 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5521 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6301 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6361 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6421 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6481 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6541 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6601 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     6661 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     6721 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     6781 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     6841 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     6901 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6961 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7021 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     7081 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     7141 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     7201 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     7261 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     7321 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     7381 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     7441 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     7501 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7561 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     7621 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7681 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     7741 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     7801 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     7861 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     7921 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     7981 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     8041 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     8101 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     8161 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     8221 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     8281 agaatagttt ttgctgtact ttctatagtg aatagagtt
//
