LOCUS       sequence-191028-       11625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and hygromycin resistance marker with 2A 
            iRFP670 from EF-1a promoter. 3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11625)
  AUTHORS   Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I, 
            Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ
  TITLE     Evaluating Synthetic Activation and Repression of 
            Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and 
            Astrocytes.
  JOURNAL   Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 
            10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27.
  PUBMED    28757163
REFERENCE   2  (bases 1 to 11625)
  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: 
            "10.1016/j.stemcr.2017.06.012"; journalName: "Stem Cell Reports"; 
            date: "2017-08-8- 8"; volume: "9"; issue: "2"; pages: "615-628"
COMMENT     Sequence Label: lentiGuide-Hygro-iRFP670
FEATURES             Location/Qualifiers
     source          1..11625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..1208
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          261..1199
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1156..1176
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1215..1224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1230..2246
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     CDS             2271..2327
                     /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"
     misc_feature    3278..3866
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3331..3351)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3749..3760)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3938..4171
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4249..4370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4286..4305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4340..4359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4410..4545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4565..4584)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4566..4584)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4594..4611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4594..4610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4603..4625)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4752..5207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4839..4858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5049..5070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5233..5337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5338..5406,5407..6198)
                     /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(5556..5575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6369..6957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6858..6877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7111..7128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7245..7266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7281..7286,7287..7304,7305..7311)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7319..7335
                     /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     7324..7346
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7343..7359
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7343..7359
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7378..7398
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7380..7398
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7426..7652
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7653..7833
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7880..8005
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8498..8731
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8916..8960
                     /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"
     promoter        9138..9386
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9138..9158
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9309..9328
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11268..11343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    11399..11516
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
       61 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga
      121 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      181 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      241 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
      301 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
      361 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
      421 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
      481 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
      541 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
      601 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
      661 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
      721 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
      781 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
      841 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
      901 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
      961 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     1021 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     1081 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     1141 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     1201 tgtcgtgacg tacggccacc atggaattca aaaagcctga actcaccgct acctctgtcg
     1261 agaagtttct gatcgaaaag tttgacagtg tatccgacct gatgcagctc tccgagggcg
     1321 aagaatctcg ggctttcagc ttcgatgtgg gagggcgtgg atatgtcctg cgggtgaata
     1381 gctgcgccga tggtttctac aaagatcgct atgtttatcg gcactttgca tccgccgctc
     1441 tccctattcc cgaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct
     1501 cccgccgtgc acagggtgtc accttgcaag acctgcctga aaccgaactg cccgctgttc
     1561 tccagcccgt cgccgaggcc atggatgcca tcgctgccgc cgatcttagc cagaccagcg
     1621 ggttcggccc attcggacct caaggaatcg gtcaatacac tacatggcgc gatttcatct
     1681 gcgctattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     1741 cctccgtcgc ccaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     1801 ggcacctcgt gcacgccgat ttcggctcca acaatgtcct gaccgacaat ggccgcataa
     1861 cagccgtcat tgactggagc gaggccatgt tcggggattc ccaatacgag gtcgccaaca
     1921 tcttcttctg gaggccctgg ttggcttgta tggagcagca gacccgctac ttcgagcgga
     1981 ggcatcccga gcttgcagga tctcctcggc tccgggctta tatgctccgc attggtcttg
     2041 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gctcagggtc
     2101 gctgcgacgc aatcgtccgg tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     2161 gaagcgctgc cgtctggacc gatggctgtg tggaagtgct cgccgatagt ggaaacagac
     2221 gccccagcac tcgtcctagg gcaaaggatc tgcagtctag aggatctggt gccacgaatt
     2281 tctcactact caagcaggcc ggtgatgtcg aggaaaaccc tggtcctatg gcccggaagg
     2341 tggacctgac cagctgcgat agagagccaa tccacatccc cggctccatc cagccatgcg
     2401 gatgtctgct ggcatgtgac gcacaggccg tgcggatcac cagaatcaca gagaacgcag
     2461 gagccttctt tggaagggag accccaaggg tgggagagct gctggccgat tacttcggag
     2521 agacagaggc acacgcactg aggaatgcac tggcacagag ctccgacccc aagagacctg
     2581 ccctgatctt cggatggagg gatggactga ccggaagaac atttgacatc tccctgcacc
     2641 ggcacgatgg cacatctatc atcgagtttg agccagcagc agcagagcag gcagacaacc
     2701 cactgaggct gacccgccag atcatcgcca gaacaaagga gctgaagagc ctggaggaga
     2761 tggcagcaag ggtgcctaga tacctgcagg ccatgctggg ctaccacaga gtgatgctgt
     2821 atagattcgc cgacgatggc tctggcatgg tcatcggcga ggccaagagg agcgacctgg
     2881 agtccttcct gggacagcac tttccagcat ccctggtgcc tcagcaggca aggctgctgt
     2941 atctgaagaa cgccatccgc gtggtgtccg attctagggg catctctagc cgcatcgtgc
     3001 cagagcacga cgcatctgga gccgccctgg atctgagctt cgcccacctg aggagcatct
     3061 ccccttgcca cctggagttt ctgcgcaata tgggcgtgtc tgccagcatg tccctgtcta
     3121 tcatcatcga cggcaccctg tggggcctga tcatctgtca ccactatgag cctagggcag
     3181 tgccaatggc acagagggtg gcagcagaga tgttcgccga ttttctgtcc ctgcacttta
     3241 cagccgccca ccaccagcgg tgacgcgtta agtcgacaat caacctctgg attacaaaat
     3301 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3361 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3421 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3481 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3541 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3601 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3661 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     3721 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3781 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3841 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     3901 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3961 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     4021 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     4081 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     4141 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt
     4201 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     4261 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4321 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4381 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     4561 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     4621 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4681 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4741 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4801 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5101 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5161 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     5221 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5581 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5881 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6481 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7081 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7141 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7201 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7261 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7321 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     7381 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     7441 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     7501 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     7561 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     7621 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     7681 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7741 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7801 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     7861 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     7921 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     7981 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     8041 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     8101 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     8161 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     8221 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     8281 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     8341 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     8401 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     8461 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     8521 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     8581 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     8641 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     8701 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     8761 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     8821 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     8881 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     8941 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     9001 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     9061 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     9121 ccccgagggg acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     9181 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     9241 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     9301 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     9361 atatcttgtg gaaaggacga aacaccggag acggttgtaa atgagcacac aaaatacaca
     9421 tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct ttttaataac
     9481 tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat aaaaatgagc
     9541 aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt aataatagct
     9601 actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg agcatcttca
     9661 aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac tgacttcatt
     9721 agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt tttaacaaat
     9781 tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt tgttaataaa
     9841 caattttcca taatacattt aacaacatgt gatccagctg ctttttttac agctttcatg
     9901 tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa taccatatca
     9961 atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt agctgttgtg
    10021 catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc ttttaataat
    10081 tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc aaattcaatt
    10141 gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa taatgtgtga
    10201 tctatatatt tgtttagttt cattttttct cctatatatt catttttaat tttaattctt
    10261 taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca cctataaaat
    10321 aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac ttatcttcta
    10381 aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg tagtcttcaa
    10441 cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat gatttcatta
    10501 atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa gtgaaagtgt
    10561 aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt ttattttgtt
    10621 ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca tctttatctc
    10681 tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca aaatttaatc
    10741 cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt aaagttttta
    10801 caatattaac tccatatttt tcatgagcga ttctatcacc caaatcactt gttacaaaac
    10861 ttgaatatag agccggattt tttggaatgc tatttaagcg ttttagattt gataattttc
    10921 aatcaattaa aattggtcct gtttgatttc catctaatct tacaaaatga ccatcatgtt
    10981 ttattgccat tccaaatctg tcagcatctg ggtcattcat aataataata tctgcatcat
    11041 gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga tttggattta
    11101 caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata acgttatatc
    11161 ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact gcgctaaaaa
    11221 taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt ttagagctag
    11281 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
    11341 tgctttttta agcttggcgt aactagatct tgagacaaat ggcagtattc atccacaatt
    11401 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
    11461 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
    11521 tttattacag ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa
    11581 gatggataaa gttttaaaca gagaggaatc tttgcagcta atgga
//
