LOCUS       sequence-429502-        9543 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express miRFP670nano P2A ERM-gCaMP6f (fluorescent proteins and 
            genetic calcium sensor, gCaMP6f for normalizing factor and sensing 
            calcium ions at cytosol exposed to ERM, respectively).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9543)
  AUTHORS   Bharat V, Durairaj AS, Vanhauwaert R, Li L, Muir CM, Chandra S, Kwak
            CS, Le Guen Y, Nandakishore P, Hsieh CH, Rensi SE, Altman RB, 
            Greicius MD, Feng L, Wang X
  TITLE     A mitochondrial inside-out iron-calcium signal reveals drug targets 
            for Parkinson's disease.
  JOURNAL   Cell Rep. 2023 Dec 26;42(12):113544. doi: 
            10.1016/j.celrep.2023.113544. Epub 2023 Dec 6.
  PUBMED    38060381
REFERENCE   2  (bases 1 to 9543)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2023-12-26"; volume: "42(12):113544. doi"; pages: " 
            10.1016/j.celrep.2023.113544. Epub 2023 Dec 6"
COMMENT     Sequence Label: pLX304_miRFP670nano-P2A-ERM-gCam6f
FEATURES             Location/Qualifiers
     source          1..9543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    590..614
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1082..1138
                     /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"
     regulatory      1253..1262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1274..1330
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1543..1564
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1670..1691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1931..1950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2545..3133
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2598..2618)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3016..3027)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3205..3438
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3516..3637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3553..3572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3607..3626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3677..3812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3832..3851)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3833..3851)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3861..3878)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3861..3877)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3870..3892)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4019..4474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4106..4125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4316..4337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4500..4604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4605..4673,4674..5465)
                     /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(4823..4842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5636..6224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6125..6144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6378..6395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6512..6533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6548..6553,6554..6571,6572..6578)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6586..6602
                     /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     6591..6613
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6610..6626
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6610..6626
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6645..6665
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6647..6665
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6693..6919
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6920..7100
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7147..7272
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7765..7998
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8183..8227
                     /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        8402..8902
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8821..8839
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             8914..9309
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9369..9486
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctctc tggctaactg tcgggatcaa caagtttgta
      601 caaaaaagca ggcttcgaag gagatagaac catggcaaac ctggacaaga tgctgaatac
      661 cacagtaaca gaggtgcggc agttcctgca ggtggacaga gtgtgcgtgt tccagtttga
      721 ggaggattat agcggagtgg tggtggtgga ggccgtggac gataggtgga tctccatcct
      781 gaagacccag gtgcgggata gatacttcat ggagacaagg ggcgaggagt attctcacgg
      841 ccgctaccag gccatcgccg acatctacac cgcaaacctg acagagtgct acagggatct
      901 gctgacacag tttcaggtga gagcaatcct ggccgtgccc atcctgcagg gcaagaagct
      961 gtggggcctg ttggtggcac accagctggc ggcccctaga cagtggcaga cctgggagat
     1021 cgactttctg aagcagcagg ccgtggtggt gggcatcgcc atccagcaga gcggaagcgg
     1081 agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctcg
     1141 tacgatgcat ccggatatgg accctgtggt cgtgctgggg ctgtgcctga gctgtctgct
     1201 gctgctgagc ctgtggaagc agtcctacgg aggcgggttg gatccaccgg tcgccaccat
     1261 ggtcgactca tcacgtcgta agtggaataa gacaggtcac gcagtcagag ctataggtcg
     1321 gctgagctca ctcgagaacg tctatatcaa ggccgacaag cagaagaacg gcatcaaggc
     1381 gaacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgcct accactacca
     1441 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcgt
     1501 gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1561 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg
     1621 gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1681 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg gcgatgccac
     1741 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1801 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1861 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg agcgcaccat
     1921 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1981 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2041 gcacaagctg gagtacaacc tgccggacca actgactgaa gagcagatcg cagaatttaa
     2101 agaggaattc tccctatttg acaaggacgg ggatgggaca ataacaacca aggagctggg
     2161 gacggtgatg cggtctctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa
     2221 tgaagtagat gccgacggtg acggcacaat cgacttccct gagttcctga caatgatggc
     2281 aagaaaaatg aaatacaggg acacggaaga agaaattaga gaagcgttcg gtgtgtttga
     2341 taaggatggc aatggctaca tcagtgcagc agagcttcgc cacgtgatga caaaccttgg
     2401 agagaagtta acagatgaag aggttgatga aatgatcagg gaagcagaca tcgatgggga
     2461 tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagcttt aagctagcgc
     2521 taaccggtgg cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     2581 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2641 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2701 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2761 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2821 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2881 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2941 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3001 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3061 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3121 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3181 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     3241 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3301 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3361 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3421 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     3481 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     3541 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3601 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3781 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     3841 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3901 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3961 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4021 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4081 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4141 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4201 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4261 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4321 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4381 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4441 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4501 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4561 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4621 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4681 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4741 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4801 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4861 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4921 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4981 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5041 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5101 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5161 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5221 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5281 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5341 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5401 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5461 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5521 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5581 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6241 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6301 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6361 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6421 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6481 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     6541 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6601 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     6661 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     6721 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     6781 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     6841 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     6901 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     6961 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7021 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7081 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     7141 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     7201 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     7261 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     7321 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     7381 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     7441 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     7501 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     7561 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     7621 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     7681 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     7741 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     7801 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     7861 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     7921 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     7981 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     8041 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     8101 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     8161 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     8221 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     8281 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     8341 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     8401 cttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     8461 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     8521 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     8581 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     8641 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     8701 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     8761 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     8821 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     8881 aatcaccgac ctctctcccc agggggtacc accatggcca agcctttgtc tcaagaagaa
     8941 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     9001 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     9061 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     9121 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     9181 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     9241 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     9301 tgggagggcc tgcagctgca gtagtaagaa ttctagatct tgagacaaat ggcagtattc
     9361 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     9421 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     9481 aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct cgagggggcc
     9541 cgg
//
