LOCUS       sequence-476966-       10672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral expression of humanized ChR2 with H134R mutation 
            fused to EYFP driven by EF1a promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10672)
  AUTHORS   Zhang F, Wang LP, Brauner M, Liewald JF, Kay K, Watzke N, Wood PG, 
            Bamberg E, Nagel G, Gottschalk A, Deisseroth K
  TITLE     Multimodal fast optical interrogation of neural circuitry.
  JOURNAL   Nature. 2007 Apr 5. 446(7136):633-9.
  PUBMED    17410168
REFERENCE   2  (bases 1 to 10672)
  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: "Nature. 
            2007 Apr 5. 446(7136):633-9."
COMMENT     Sequence Label: pLenti-EF1a-hChR2(H134R)-EYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..10672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        324..1502
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          555..1493
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1450..1470
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    1550..1667
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      1810..1819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1816..2742
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2752..2761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2758..2760,2761..2763,2764..3477)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2803..2824)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3064..3083)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3411..3432
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3495..4083
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3548..3568)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3966..3977)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    4251..4269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4293..4311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4335..4353
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4377..4395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4419..4437
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4461..4479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        4508..4722
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     4643..4662
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     LTR             4741..4921
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     CDS             4947..4988
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4998..5015
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5038..5055)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5044..5268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5314..5742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5401..5420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5611..5632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5751..5771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5756..6085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5936..6071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5998..6017
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6133..6180
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6199..6597
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6755..6876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6792..6811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6846..6865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6925..6941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6925..6941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6938..6960)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6949..6965
                     /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(6973..6979,6980..6997,6998..7003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7018..7039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7156..7173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7316..7904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7396..7415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8075..8866,8867..8935))
                     /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     8698..8717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8936..9040)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        9077..9303
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9304..9484
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9531..9656
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10149..10382
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10567..10611
                     /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 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
       61 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
      121 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
      181 acggatctcg acggtatcga tcacgagact agcctcgacc atcgcgatgt cgacgataag
      241 ctttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct
      301 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      361 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
      421 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      481 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      541 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
      601 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
      661 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
      721 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
      781 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
      841 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
      901 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
      961 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     1021 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     1081 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     1141 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     1201 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     1261 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     1321 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     1381 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     1441 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     1501 gaggaatttc gacatttaaa tttaattaat ctcgacggta tcggttaact tttaaaagaa
     1561 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     1621 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga
     1681 ctagcctcga ggtttaaact acgggatctt cgaaggccta agcttacgcg cgcgtcctag
     1741 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
     1801 gaaaccggtg ccaccatgga ctatggcggc gctttgtctg ccgtcggacg cgaacttttg
     1861 ttcgttacta atcctgtggt ggtgaacggg tccgtcctgg tccctgagga tcaatgttac
     1921 tgtgccggat ggattgaatc tcgcggcacg aacggcgctc agaccgcgtc aaatgtcctg
     1981 cagtggcttg cagcaggatt cagcattttg ctgctgatgt tctatgccta ccaaacctgg
     2041 aaatctacat gcggctggga ggagatctat gtgtgcgcca ttgaaatggt taaggtgatt
     2101 ctcgagttct tttttgagtt taagaatccc tctatgctct accttgccac aggacaccgg
     2161 gtgcagtggc tgcgctatgc agagtggctg ctcacttgtc ctgtcatcct tatccgcctg
     2221 agcaacctca ccggcctgag caacgactac agcaggagaa ccatgggact ccttgtctca
     2281 gacatcggga ctatcgtgtg gggggctacc agcgccatgg caaccggcta tgttaaagtc
     2341 atcttctttt gtcttggatt gtgctatggc gcgaacacat tttttcacgc cgccaaagca
     2401 tatatcgagg gttatcatac tgtgccaaag ggtcggtgcc gccaggtcgt gaccggcatg
     2461 gcatggctgt ttttcgtgag ctggggtatg ttcccaattc tcttcatttt ggggcccgaa
     2521 ggttttggcg tcctgagcgt ctatggctcc accgtaggtc acacgattat tgatctgatg
     2581 agtaaaaatt gttgggggtt gttgggacac tacctgcgcg tcctgatcca cgagcacata
     2641 ttgattcacg gagatatccg caaaaccacc aaactgaaca tcggcggaac ggagatcgag
     2701 gtcgagactc tcgtcgaaga cgaagccgag gccggagccg tgccagcggc cgccaccatg
     2761 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2821 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2881 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2941 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     3001 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3061 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3121 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3181 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     3241 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3301 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3361 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3421 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatag
     3481 actagtcata tgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3541 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3601 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3661 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3721 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3781 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3841 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     3901 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3961 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4021 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4081 cgcatcggta cgtatggcca ggtaccttta agaccaatga cttacaaggc agctgtagat
     4141 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     4201 caagatggga tcaattcacc atgctagtgg atccccctcg agtttaccac tccctatcag
     4261 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     4321 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     4381 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
     4441 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cggggctgca
     4501 ggaattcgaa cgctgacgtc atcaacccgc tccaaggaat cgcgggccca gtgtcactag
     4561 gcgggaacac ccagcgcgcg tgcgccctgg caggaagatg gctgtgaggg acaggggagt
     4621 ggcgccctgc aatatttgca tgtcgctatg tgttctggga aatcaccata aacgtgaaat
     4681 gtctttggat ttgggaatct tataagttct gtatgagacc acagatctaa gcttatcgat
     4741 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4801 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4861 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4921 agcatctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     4981 attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     5041 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     5101 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     5161 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     5221 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     5281 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     5341 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5401 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5461 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5521 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5581 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5641 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5701 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     5761 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5821 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5881 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5941 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6001 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     6061 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     6121 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     6181 aaggtgagga actaaaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     6241 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     6301 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     6361 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     6421 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     6481 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     6541 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagca
     6601 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     6661 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6721 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6781 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6841 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6901 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7261 aaagaacatg tgagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8161 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8221 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8281 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8341 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8401 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8461 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8521 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8581 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8641 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8701 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8761 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8821 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8881 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8941 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9001 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9061 cacctgacgt ccttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     9121 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     9181 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     9241 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     9301 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     9361 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     9421 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     9481 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     9541 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     9601 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     9661 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     9721 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     9781 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     9841 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     9901 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     9961 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
    10021 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
    10081 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
    10141 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
    10201 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
    10261 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
    10321 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
    10381 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
    10441 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
    10501 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
    10561 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
    10621 tttaacataa caaattggct gtggtatata aaattattca taatgatagt ag
//
