LOCUS       sequence-340562-        9690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of G-alpha-s fused to miRFP670-eDHFR(69K6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9690)
  AUTHORS   Suzuki S, Nakamura A, Hatano Y, Yoshikawa M, Yoshii T, Sawada S, 
            Atsuta-Tsunoda K, Aoki K, Tsukiji S
  TITLE     A chemogenetic platform for controlling plasma membrane signaling 
            and synthetic signal oscillation.
  JOURNAL   Cell Chem Biol. 2022 Sep 15;29(9):1446-1464.e10. doi: 
            10.1016/j.chembiol.2022.06.005. Epub 2022 Jul 13.
  PUBMED    35835118
REFERENCE   2  (bases 1 to 9690)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2022.06.005"; journalName: "Cell Chem Biol"; 
            date: "2022-09-15- 15"; volume: "29"; issue: "9"; pages: 
            "1446-1464.e10"
COMMENT     Sequence Label: pPBpuro-miRFP670-eDHFR(69K6)-G-alpha-s
FEATURES             Location/Qualifiers
     source          1..9690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1668..2684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2608..2630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2692..2711
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2757..2766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2763..3707
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     misc_feature    5596..6145
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5740..5757)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5967..5986
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6148..6747
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6148..6167)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6644..6664
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6774..6791)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6780..7004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7051..7067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7075..7091
                     /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(7099..7105,7106..7123,7124..7129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7144..7165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7399..7461
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7502..7522)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7502..7520)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7541..7557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7565..7581
                     /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(7589..7595,7596..7613,7614..7619))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7634..7655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7772..7789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7943..8531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8023..8042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8702..9493,9494..9562))
                     /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     9325..9344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9563..9667)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     1441 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     1501 gggggggggg gsgcgcgccr gggggggggg gggggggggg gggggggggs ggggsgrggs
     1561 ggrgaggkgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1621 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1681 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1741 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1801 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1861 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1921 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1981 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     2041 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     2101 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     2161 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     2221 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     2281 ccggggrggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     2341 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     2401 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     2461 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     2521 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2581 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2641 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     2701 tggttattgt gctgtctcat cattttggca aagaattggg atcctcacgt caattggcca
     2761 ccatggtagc aggtcatgcc tctggcagcc ccgcattcgg gaccgcctct cattcgaatt
     2821 gcgaacatga agagatccac ctcgccggct cgatccagcc gcatggcgcg cttctggtcg
     2881 tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa cgccgcggaa tttctgaatc
     2941 tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg cgatctgttg atcaagatcc
     3001 tgccgcatct cgatcccacc gccgaaggca tgccggtcgc ggtgcgctgc cggatcggca
     3061 atccctctac ggagtactgc ggtctgatgc atcggcctcc ggaaggcggg ctgatcatcg
     3121 aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac gctggcgccg gcgctggagc
     3181 ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga caccgtgctg ctgtttcagc
     3241 agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga tgagcaaggc cacggcctgg
     3301 tattctccga gtgccatgtg cctgggctcg aatcctattt cggcaaccgc tatccgtcgt
     3361 cgactgtccc gcagatggcg cggcagctgt acgtgcggca gcgcgtccgc gtgctggtcg
     3421 acgtcaccta tcagccggtg ccgctggagc cgcggctgtc gccgctgacc gggcgcgatc
     3481 tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgtg ccatctgcag ttcctgaagg
     3541 acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt cggcggcaag ctgtggggcc
     3601 tggttgtctg tcaccattat ctgccgcgct tcatccgttt cgagctgcgg gcgatctgca
     3661 aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct tgagagcctc tacaagtctg
     3721 ggtccgcagc cagtagcgcc ggaagtatga tcagtctgat tgcggcgtta gcggtagatc
     3781 gcgttatcgg catggaaaac gccatgccgt ggaacctgcc tgccgatctc gcctggttta
     3841 aacgcaacac cttaaataaa cccgtgatta tgggccgcca tacctgggaa tcaatcggtc
     3901 gtccgttgcc aggacgcaaa aatattatcc tcagcagtca accgggtacg gacaaaaaaa
     3961 agaaaaagaa agatcgcgta acgtgggtga agtcggtgga tgaagccatc gcggcgtgtg
     4021 gtgacgtacc agaaatcatg gtgattggcg gcggtcgcgt ttatgaacag ttcttgccaa
     4081 aagcgcaaaa actgtatctg acgcatatcg acgcagaagt ggaaggcgac acccatttcc
     4141 cggattacga gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc
     4201 agaactctca cagctattgc tttgagattc tggagcggcg gctgtacaag tccggactca
     4261 gatctcgaca aggtagtggt gctggctctg gtgctggtag tggcgctggt tccggtgctg
     4321 gctctggcgc gcctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
     4381 gtatgggcag cctcggcaac agtaagaccg aggaccagcg caacgaggag aaggcgcagc
     4441 gcgaggccaa caaaaagatc gagaagcagc tgcagaagga caagcaggtc taccgggcca
     4501 cgcaccgcct gctgctgctg ggtgctggag agtctggcaa aagcaccatt gtgaagcaga
     4561 tgaggatcct acatgttaat gggtttaacg gagacgagaa ggccaccaaa gtgcaggaca
     4621 tcaaaaacaa cctgaaggag gccattgaaa ccattgtggc cgccatgagc aacctggtgc
     4681 cccccgtgga gctggccaac cctgagaacc agttcagagt ggactacatt ctgagcgtga
     4741 tgaacgtgcc aaactttgac ttcccacctg aattctatga gcatgccaag gctctgtggg
     4801 aggatgaggg agttcgtgcc tgctacgagc gctccaacga gtaccagctg atcgactgtg
     4861 cccagtactt cctggacaag attgatgtga tcaagcaggc cgactacgtg ccaagtgacc
     4921 aggacctgct tcgctgccgc gtcctgacct ctggaatctt tgagaccaag ttccaggtgg
     4981 acaaagtcaa cttccacatg ttcgatgtgg gcggccagcg cgatgaacgc cgcaagtgga
     5041 tccagtgctt caatgatgtg actgccatca tcttcgtggt ggccagcagc agctacaaca
     5101 tggtcatccg ggaggacaac cagaccaacc gtctgcagga ggctctgaac ctcttcaaga
     5161 gcatctggaa caacagatgg ctgcgtacca tctctgtgat cctcttcctc aacaagcaag
     5221 atctgcttgc tgagaaggtc ctcgctggga aatcgaagat tgaggactac tttccagagt
     5281 tcgctcgcta caccactcct gaggatgcga ctcccgagcc cggagaggac ccacgcgtga
     5341 cccgggccaa gtacttcatc cgggatgagt ttctgagaat cagcactgct agtggagatg
     5401 gacgtcacta ctgctaccct cactttacct gcgccgtgga cactgagaac atccgccgtg
     5461 tcttcaacga ctgccgtgac atcatccagc gcatgcatct tcgccaatac gagctgctct
     5521 aaaccgggat aagtcaacta acttaagcta gcaacggttt ccctctagcg ggatcaattc
     5581 cgcccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     5641 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     5701 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     5761 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     5821 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     5881 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     5941 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     6001 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     6061 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     6121 ttttcctttg aaaaacacga taataccatg accgagtaca agcccacggt gcgcctcgcc
     6181 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     6241 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     6301 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6361 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6421 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6481 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     6541 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6601 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6661 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6721 tgcatgaccc gcaagcccgg tgcctgatcg atgatctaga gctcgctgat cagcctcgac
     6781 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6841 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6901 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6961 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     7021 aacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7201 aaacctgtcg tgccagcgga tcttaaaagt tttgttactt tatagaagaa attttgagtt
     7261 tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat ttattattag
     7321 tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat aaacctcgat
     7381 atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac
     7441 aatatgatta tctttctagg gttaatctag tatacgcgtt taattaatcc agcttttgtt
     7501 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7681 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7741 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7801 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7861 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8341 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8701 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8761 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8821 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8881 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8941 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9001 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9061 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9121 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9181 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9241 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9301 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9361 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9421 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9481 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9541 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9601 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9661 ttccgcgcac atttccccga aaagtgccac
//
