LOCUS       sequence-123118-        9158 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pPBC-LG3-tdT expresses PiggyBac inverted terminal repeat-flanked 
            Lck-GCaMP3 and tdTomato under the control of the CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9158)
  AUTHORS   Gee JM, Gibbons MB, Taheri M, Palumbos S, Morris SC, Smeal RM, Flynn
            KF, Economo MN, Cizek CG, Capecchi MR, Tvrdik P, Wilcox KS, White JA
  TITLE     Imaging activity in astrocytes and neurons with genetically encoded 
            calcium indicators following in utero electroporation.
  JOURNAL   Front Mol Neurosci. 2015 Apr 15;8:10. doi: 10.3389/fnmol.2015.00010.
            eCollection 2015.
  PUBMED    25926768
REFERENCE   2  (bases 1 to 9158)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Mol 
            Neurosci."; date: "2015-04-15"; volume: "8:10. doi"; pages: " 
            10.3389/fnmol.2015.00010. eCollection 2015"
COMMENT     Sequence Label: pPBC-LG3-tdT
FEATURES             Location/Qualifiers
     source          1..9158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        451..469
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     451..468
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    535..556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(571..576,577..594,595..601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    609..625
                     /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     614..636
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     633..649
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     633..649
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        670..688
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   871..933
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1225..1604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1607..1882
                     /label=chicken beta-actin promoter
     intron          1883..2891
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2815..2837
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2899..2918
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2964..2973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3087..3098,3099..3116,3117..3119,3120..3152,3153..3155,
                     3156..3179,3180..3206,3207..3263,3264..3269,3270..3539,
                     3540..3557,3558..3992,3993..3995,3996..3998,3999..4442)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(3087..3098,3099..3116,3117..3119,3120..3152,3153..3155,
                     3156..3179,3180..3206,3207..3263,3264..3269,3270..3539,
                     3540..3557,3558..3992,3993..3995,3996..3998,3999..4442)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      3186..3195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3476..3497
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3603..3624)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3864..3883)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4463..5036
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4486..5038
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4630..4647)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4857..4876
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5053..6483
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(6665..6684)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6709..6756
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     repeat_region   7104..7138
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7262..7281)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7263..7281)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7368..7385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7539..8127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7619..7638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8298..9089,9090..9158))
                     /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     8921..8940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
      421 atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa ctcgacctcg
      481 aggctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      541 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      601 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      661 caagctcgaa attaaccctc actaaaggga acaaaagctg gagctcgtct ttgatcaaaa
      721 cgcaaatcga cgaaaatgtg tcggacaata tcaagtcgat gagcgaaaaa ctaaaaaggc
      781 tagaatacga caatctcaca gacagcgttg agatatacgg tattcacgac agcaggctga
      841 ataataaaaa aattagaaac tattatttaa ccctagaaag ataatcatat tgtgacgtac
      901 gttaaagata atcatgcgta aaattgacgc atgtgtttta tcgatctgta tatcgaggtt
      961 tatttattaa tttgaataga tattaagttt tattatattt acacttacat actaataata
     1021 aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa
     1081 tttcttctat aaagtaacaa aacttttaaa cattctctct tttacaaaaa taaacttatt
     1141 ttgtacttta aaaacagtca tgttgtatta taaaataagt aattagctta acttatacat
     1201 aatagaaaca aattatactt attagacatt gattattgac tagttattaa tagtaatcaa
     1261 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1321 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1381 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggac tatttacggt
     1441 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1501 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1561 ctacttggca gtacatctac gtattagtca tcgctattac catgggtcga ggtgagcccc
     1621 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     1681 attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg
     1741 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
     1801 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     1861 aaagcgaagc gcgcggcggg cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg
     1921 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     1981 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc
     2041 ttttctgtgg ctgcgtgaaa gccttaaagg gctccgggag ggccctttgt gcggggggga
     2101 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct
     2161 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc
     2221 gaggggagcg cggccggggg cggtgccccg cggtgcgggg gggctgcgag gggaacaaag
     2281 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc
     2341 tgtaaccccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     2401 gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
     2461 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
     2521 ggccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     2581 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg
     2641 gaggcgccgc cgcaccccct ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag
     2701 gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag
     2761 cctcggggct gccgcagggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     2821 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     2881 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     2941 ctagcgctac cggactcaga tctgccacca tgggctgtgg ctgcagctca aaccctgaag
     3001 atgactggat ggagaacatt gacgtgtgtg agaactgcca ttatcccccg ccgaagcttc
     3061 gaattgatcc accggatctc gccaccatgc ggggttctca tcatcatcat catcatggta
     3121 tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg
     3181 atctcgccac catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca
     3241 gagctatagg tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga
     3301 acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     3361 cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3421 actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg
     3481 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     3541 gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3601 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg
     3661 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3721 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3781 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc
     3841 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3901 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3961 gcaacatcct ggggcacaag ctggagtaca acacgcgtga ccaactgact gaagagcaga
     4021 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa
     4081 ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc
     4141 aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc
     4201 tgacaatgat ggcaagaaaa atgaaagaca cagacagtga agaagaaatt agagaagcgt
     4261 tccgtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga
     4321 tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag
     4381 acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt
     4441 gagcggccgc ctagcggatc tgcccctctc cctccccccc ccctaacgtt actggccgaa
     4501 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     4561 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     4621 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     4681 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     4741 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     4801 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     4861 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     4921 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     4981 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     5041 gccacaacat atatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
     5101 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     5161 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     5221 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     5281 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     5341 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     5401 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     5461 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     5521 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     5581 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     5641 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     5701 cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc
     5761 agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc
     5821 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     5881 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     5941 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     6001 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     6061 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     6121 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     6181 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     6241 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     6301 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     6361 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     6421 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     6481 taagcggccg cttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6541 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6601 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     6661 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     6721 gacagaataa aacgcacggt gttgggtcgt ttgttcggcg cgccatcgaa gcagctccag
     6781 cctacacaat cgctcaagac gtgtaatgct tttattatat attagtcacg atatctataa
     6841 caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt
     6901 atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac
     6961 gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga
     7021 gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa
     7081 gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt
     7141 aaaaaagatt tgcgctttac tcgacctaaa ctttaaacac gtcatagaat cttcgtttga
     7201 caaaaaccac attgtggggt accgagctcg aattcatcga tgatatcaga tctgccggtc
     7261 tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc
     7321 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7381 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7441 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8281 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8341 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8401 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8461 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8521 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8581 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8641 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8701 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8761 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8821 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8881 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8941 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9001 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9061 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9121 aagggaataa gggcgacacg gaaatgttga atactcat
//
