LOCUS       sequence-455550-        8941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle-entry vector with dsRed flanked by loxP sites and spCas9-GFP 
            for Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8941)
  AUTHORS   Siniscalco AM, Perera RP, Greenslade JE, Veeravenkatasubramanian H, 
            Masters A, Doll HM, Raj B
  TITLE     Barcoding Notch signaling in the developing brain.
  JOURNAL   Development. 2024 Nov 22:dev.203102. doi: 10.1242/dev.203102.
  PUBMED    39575683
REFERENCE   2  (bases 1 to 8941)
  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: 
            "Development. 2024 Nov 22:dev.203102. doi: 10.1242/dev.203102."
COMMENT     Sequence Label: p155ME-loxP-dsR-loxP-Cas9-t2A-GFP
FEATURES             Location/Qualifiers
     source          1..8941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             573..581
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             676..696
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             712..765
                     /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(772..774,775..777,778..1491)
                     /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(772..774,775..777,778..1488)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(817..838)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1078..1097)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1425..1446
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1528..1548)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1528..1546)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(1575..1674)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1692..1710)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1715..1731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1715..1731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1833..1852)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1844..2653
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1911..1930)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2800..3388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3289..3308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3542..3559
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3718..3745)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3837..3923)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3972..3994
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3987..4003
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4019..4118
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     4120..4136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4146..4164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(4203..4236)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4243..4252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4249..4926
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(4429..4449)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4841..4864
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    5047..5168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5084..5103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5138..5157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5312..5333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    complement(5422..5455)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             5474..5494
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5501..8941,1..660)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes MGAS15252 Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 gtggagaagg gcaaaagcaa gaaactgaaa tctgtcaagg agctgctggg tatcacaatt
       61 atggagagaa gctctttcga gaagaaccca atcgattttc tggaggccaa aggttataag
      121 gaagtgaaga aagacctgat cattaaactg cccaagtaca gtctgtttga gctggaaaac
      181 ggcaggaaac gcatgctggc aagcgctgga gagctgcaga aaggcaatga actggccctg
      241 ccttctaagt acgtgaactt cctgtatctg gcaagccact acgagaagct gaaaggatct
      301 ccagaggata acgaacagaa acagctgttt gtggagcagc acaagcatta tctggacgag
      361 atcattgaac agattagcga gttctctaaa agagtgatcc tggccgacgc aaatctggat
      421 aaggtcctgt ctgcttacaa caaacacaga gataagccca tcagggagca ggccgaaaat
      481 atcattcatc tgttcactct gacaaacctg ggcgcacctg cagctttcaa gtacttcgac
      541 actacaatcg atagaaagag gtacacctcc actaaggagg tgctggacgc tacactgatc
      601 catcagagta ttaccggcct gtacgaaaca aggattgacc tgtctcagct gggtggcgac
      661 tcaccggtgg gatcccctaa gaagaagaga aaggtgaggt ctggtggcgg agagggcaga
      721 ggcagtctgc tgacatgcgg tgacgtggaa gagaatcccg gcccttctag aatggtgagc
      781 aagggcgaag agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      841 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      901 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      961 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1021 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1081 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1141 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1201 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1261 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1321 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1381 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1441 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agcggccgcc
     1501 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     1561 atcatggtca tagcacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1621 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1681 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1741 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1801 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1861 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1921 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     1981 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2041 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2101 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2161 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2221 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2281 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2341 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2401 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2461 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2521 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2581 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2641 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2701 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2941 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3421 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3481 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3541 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3601 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3661 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     3721 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     3781 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     3841 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     3901 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     3961 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
     4021 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
     4081 cttttttata atgccaactt tgtacaaaaa agcaggcttg taaaacgacg gccagtgagc
     4141 gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc gagggccggc
     4201 ctataacttc gtatagcata cattatacga agttatccgg tcgccaccat ggcctcctcc
     4261 gagaacgtca tcaccgagtt catgcgcttc aaggtgcgca tggagggcac cgtgaacggc
     4321 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcca caacaccgtg
     4381 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     4441 ttccagtacg gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag
     4501 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga actttgagga cggcggcgtg
     4561 gcgaccgtga cccaggactc ctccctgcag gacggctgct tcatctacaa ggtgaagttc
     4621 atcggcgtga acttcccctc cgacggcccc gtgatgcaga agaagaccat gggctgggag
     4681 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagac ccacaaggcc
     4741 ctgaagctga aggacggcgg ccactacctg gtggagttca agtccatcta catggccaag
     4801 aagcccgtgc agctgcccgg ctactactac gtggacgcca agctggacat cacctcccac
     4861 aacgaggact acaccatcgt ggagcagtac gagcgcaccg agggccgcca ccacctgttc
     4921 ctgtagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     4981 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5041 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5101 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5161 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5221 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5281 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5341 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5401 tgaaccatca ccctaatcaa gataacttcg tatagcatac attatacgaa gttatcccgt
     5461 catggcttct ccacctaaga agaagagaaa ggtgggaagc atggataaga agtatagcat
     5521 cggcctggat attggaacta actccgtggg ttgggcagtg attacagacg actacaaggt
     5581 ccctagcaag aaatttaagg tgctgggtaa caccgacagg cacagcatca agaaaaatct
     5641 gattggagcc ctgctgttcg gttctggaga gactgccgaa gcaacacgcc tgaaaagaac
     5701 agcaagaagg cgctatacca gaaggaagaa tagaatctgt tacctgcagg agattttctc
     5761 taacgaaatg gctaaggtgg acgattcatt ctttcatagg ctggaggaaa gtttcctggt
     5821 cgaggaagat aagaaacacg agcgccatcc tatctttgga aacattgtgg acgaggtcgc
     5881 ctatcacgaa aaatacccaa ccatctatca tctgcgcaag aaactggctg actctactga
     5941 taaagccgac ctgagactga tctatctggc tctggcccac atgattaagt tcaggggtca
     6001 ttttctgatc gagggcgatc tgaaccccga caattccgat gtggacaagc tgttcatcca
     6061 gctggtccag atttacaatc agctgtttga ggaaaaccct attaatgctt ccagagtgga
     6121 cgcaaaagct atcctgtcag ccaggctgtc caagtcacgc agactggaga acctgattgc
     6181 acagctgccc ggagaaaaga ggaacggtct gtttggaaat ctgatcgctc tgagtctggg
     6241 cctgactcct aacttcaaaa gcaattttga tctggctgag gacgccaaac tgcagctgtc
     6301 aaaggacaca tatgacgatg acctggataa cctgctggca cagatcggag atcagtacgc
     6361 tgacctgttc ctggctgcca aaaatctgtc cgacgcaatc ctgctgtcag atattctgag
     6421 agtgaacagc gagattacaa aagcacctct gagtgccagc atgatcaaga gatatgacga
     6481 gcaccatcag gatctgaccc tgctgaaggc tctggtcagg cagcagctgc cagagaagta
     6541 caaggaaatt ttctttgatc agtccaagaa cggctacgcc ggttatatcg acggaggcgc
     6601 atcacaggag gaattctaca agtttatcaa acctattctg gagaagatgg acggaactga
     6661 ggaactgctg gtgaaactga atagagagga cctgctgagg aagcagcgca catttgataa
     6721 cggttccatc ccacaccaga ttcatctggg agagctgcac gctatcctga ggcgccagga
     6781 agacttctac ccctttctga aagataaccg cgagaagatc gaaaaaattc tgaccttcag
     6841 aatcccttac tatgtgggtc cactggctcg cggaaacagc agatttgcct ggatgactcg
     6901 caaatccgag gaaaccatta ctccttggaa cttcgaggaa gtggtcgata agggcgcctc
     6961 tgcacagtcc ttcatcgaga gaatgactaa ttttgacaaa aacctgccca atgagaaagt
     7021 gctgcctaag cactccctgc tgtacgagta tttcactgtc tataacgaac tgacaaaggt
     7081 gaaatacgtc accgagggca tgagaaagcc agccttcctg tcaggagagc agaagaaagc
     7141 aatcgtggat ctgctgttta aaaccaacag gaaagtgact gtcaagcagc tgaaggagga
     7201 ctacttcaag aaaattgaat gcttcgattc cgtggagatc agcggagtcg aagacagatt
     7261 taacgcaagc ctgggcgctt accacgatct gctgaagatc attaaggata aagacttcct
     7321 ggacaacgag gaaaatgagg atatcctgga agacattgtg ctgacactga ccctgtttga
     7381 ggacagagga atgatcgagg aaagactgaa aacctatgct catctgttcg atgacaaggt
     7441 gatgaaacag ctgaagagaa ggcgctacac tggctggggt agactgagca ggaagctgat
     7501 caacggcatt agggataaac agtcaggaaa gacaatcctg gactttctga aaagtgatgg
     7561 cttcgccaac cgcaatttta tgcagctgat tcacgatgac agtctgacct tcaaagagga
     7621 catccagaag gctcaggtgt ctggacaggg ccactccctg catgagcaga ttgcaaacct
     7681 ggctggaagc ccagccatca agaaaggcat tctgcagaca gtgaaaatcg tcgatgagct
     7741 ggtgaaagtc atgggccata agcccgaaaa catcgtgatt gagatggctc gcgaaaatca
     7801 gacaacccag aagggtcaga agaacagtag agagaggatg aaaagaatcg aggaaggcat
     7861 taaggagctg ggtagccaga tcctgaaaga gcacccagtg gaaaacacac agctgcagaa
     7921 tgagaagctg tatctgtact atctgcagaa tggaagagat atgtacgtgg accaggagct
     7981 ggatattaac aggctgtctg attacgacgt ggatcatatc gtcccccaga gtttcatcaa
     8041 agatgacagc attgacaaca aggtgctgac caggtccgac aaaaacagag gaaaatcaga
     8101 taatgtccct agtgaggaag tggtcaagaa aatgaagaac tactggagac agctgctgaa
     8161 tgccaaactg atcactcaga ggaagtttga taacctgaca aaagcagagc gcggtggact
     8221 gtcagaactg gacaaagctg gattcatcaa gaggcagctg gtggaaacac gccagatcac
     8281 taaacacgtc gcacagattc tggatagtcg catgaacaca aagtacgatg agaatgacaa
     8341 actgatcaga gaagtgaagg tcattaccct gaagagtaaa ctggtcagcg actttaggaa
     8401 agatttccag ttttataagg tccgcgagat taacaattat caccatgccc atgacgcata
     8461 cctgaacgcc gtggtcggta ccgcactgat caagaaatac ccaaaactgg agagcgaatt
     8521 cgtgtacgga gactataagg tgtacgatgt cagaaaaatg atcgccaagt ccgagcagga
     8581 aattggaaaa gctactgcca agtatttctt ttactcaaac atcatgaatt tctttaagac
     8641 agagatcacc ctggccaatg gagaaatccg caaaaggccc ctgattgaga caaacggaga
     8701 gacaggcgaa atcgtgtggg acaaaggcag agattttgca accgtgagga aggtcctgag
     8761 catgcctcaa gtgaatatcg tcaagaaaac tgaggtgcag acaggcggtt tctcaaaaga
     8821 aagtattctg ccaaaacgca actctgataa gctgatcgct agaaagaaag actgggaccc
     8881 taagaagtat ggaggctttg actctcccac tgtggcatac tccgtcctgg tggtcgctaa
     8941 g
//
