LOCUS       sequence-90416-e       10696 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses the calcium sensor GCaMP5G in the mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10696)
  AUTHORS   Venkatachalam V, Cohen AE
  TITLE     Imaging GFP-Based Reporters in Neurons with Multiwavelength 
            Optogenetic Control.
  JOURNAL   Biophys J. 2014 Oct 7;107(7):1554-63. doi: 
            10.1016/j.bpj.2014.08.020.
  PUBMED    25296307
REFERENCE   2  (bases 1 to 10696)
  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"; doi: 
            "10.1016/j.bpj.2014.08"; journalName: "Biophys J"; date: "2014-10-7-
            7"; volume: "107"; issue: "7"; pages: "1554-63"
COMMENT     Sequence Label: VV073: 1xCox8 - GCaMP5G in fubi
FEATURES             Location/Qualifiers
     source          1..10696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2602..3890
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             3906..3980
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(4014..4019,4020..4037,4038..4040,4041..4073,4074..4076,
                     4077..4100,4101..4127,4128..4184,4185..4190,4191..4460,
                     4461..4478,4479..4913,4914..4919,4920..5363)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="GSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKT
                     GHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSL
                     GQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGNG
                     YISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(4014..4019,4020..4037,4038..4040,4041..4073,4074..4076,
                     4077..4100,4101..4127,4128..4184,4185..4190,4191..4460,
                     4461..4478,4479..4913,4914..4919,4920..5363)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="GSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKT
                     GHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSL
                     GQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGNG
                     YISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      4107..4116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4397..4418
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4524..4545)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4785..4804)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5388..5976
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5441..5461)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5859..5870)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5979..5995)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5980..5996)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6501..6681
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6707..6724)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6713..6937
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6983..7411
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7070..7089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7280..7301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7420..7440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7425..7754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7605..7740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7667..7686
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7802..7849
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7868..8242
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8372..8493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8409..8428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8463..8482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8542..8558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8542..8558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8555..8577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8566..8582
                     /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(8590..8596,8597..8614,8615..8620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8635..8656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8773..8790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8944..9532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9024..9043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9703..10494,10495..10563))
                     /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     10326..10345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10564..10668)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acattatggc cttaggtcac ttcatctcca tggggttctt
     2641 cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag
     2701 ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat
     2761 gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca
     2821 atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg
     2881 gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga
     2941 agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg
     3001 aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga
     3061 cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac
     3121 ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg
     3181 gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa
     3241 agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt
     3301 aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga
     3361 tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc
     3421 cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa
     3481 aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag
     3541 tttgttcaca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
     3601 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
     3661 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
     3721 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
     3781 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
     3841 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg
     3901 ccaccatgtc cgtcctgacg ccgctgctgc tgcggggctt gacaggctcg gcccggcggc
     3961 tcccagtgcc gcgcgccaag atccattcgt tgggtgaccc gccagtcgct agcggttctc
     4021 atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc
     4081 tgtacgacga tgacgataag gatctcgcca ccatggtcga ctcatcacgt cgtaagtgga
     4141 ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag aacgtctata
     4201 tcaaggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     4261 aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc
     4321 ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca
     4381 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4441 gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc
     4501 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     4561 tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4621 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4681 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4741 ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac ggcaactaca
     4801 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4861 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacctgccgg
     4921 accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta tttgacaagg
     4981 acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct ctggggcaga
     5041 accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac ggtgacggca
     5101 caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac acagacagtg
     5161 aagaagaaat tagagaagcg ttccgtgtgt ttgataagga tggcaatggc tacatcagtg
     5221 cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat gaagaggttg
     5281 atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac gaagagtttg
     5341 tacaaatgat gacagcgaag tgagaattcg atatcaagct tatcgataat caacctctgg
     5401 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5461 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5521 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5581 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5641 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5701 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5761 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     5821 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5881 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5941 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc
     6001 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     6061 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     6121 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6181 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     6241 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
     6301 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
     6361 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     6421 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
     6481 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6541 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6601 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6661 cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct
     6721 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6781 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6841 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     6901 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     6961 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     7021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     7081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     7141 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     7201 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     7261 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     7321 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     7381 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     7441 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     7501 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     7561 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     7621 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7681 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7741 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     7801 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     7861 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     7921 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     7981 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     8041 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     8101 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     8161 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     8221 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     8281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     8341 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     8401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     8461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     8521 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     8641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     9001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     9061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     9121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     9181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    10021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    10081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    10141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10681 aaaagtgcca cctgac
//
