LOCUS       sequence-340560-        8880 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of p85iSH2 fused to mNeonGreen-eDHFR(69K6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8880)
  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 8880)
  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"; 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-mNG-eDHFR(69K6)-p85iSH2
FEATURES             Location/Qualifiers
     source          1..8880
                     /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..3470
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3018..3027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4786..5335
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4930..4947)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5157..5176
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5338..5937
                     /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(5338..5357)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5834..5854
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5964..5981)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5970..6194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6241..6257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6265..6281
                     /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(6289..6295,6296..6313,6314..6319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6334..6355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6589..6651
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6692..6712)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6692..6710)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6731..6747)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6755..6771
                     /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(6779..6785,6786..6803,6804..6809))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6824..6845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6962..6979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7133..7721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7213..7232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7892..8683,8684..8752))
                     /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     8515..8534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8753..8857)
                     /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 ggrgrggkgs 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 ccatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca catgagttac
     2821 acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc accggcaatc
     2881 caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc cagttctccc
     2941 cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc taccctgacg
     3001 ggatgtcgcc tttccaggcc gccatggtag atggctccgg ctaccaagtc catcgcacaa
     3061 tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac gagggaagcc
     3121 acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt cctgtgatga
     3181 ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc aacgacaaaa
     3241 ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc taccggagca
     3301 ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg aagaaccagc
     3361 cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc aacttcaagg
     3421 agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctctacaag tctgggtccg
     3481 cagccagtag cgccggaagt atgatcagtc tgattgcggc gttagcggta gatcgcgtta
     3541 tcggcatgga aaacgccatg ccgtggaacc tgcctgccga tctcgcctgg tttaaacgca
     3601 acaccttaaa taaacccgtg attatgggcc gccatacctg ggaatcaatc ggtcgtccgt
     3661 tgccaggacg caaaaatatt atcctcagca gtcaaccggg tacggacaaa aaaaagaaaa
     3721 agaaagatcg cgtaacgtgg gtgaagtcgg tggatgaagc catcgcggcg tgtggtgacg
     3781 taccagaaat catggtgatt ggcggcggtc gcgtttatga acagttcttg ccaaaagcgc
     3841 aaaaactgta tctgacgcat atcgacgcag aagtggaagg cgacacccat ttcccggatt
     3901 acgagccgga tgactgggaa tcggtattca gcgaattcca cgatgctgat gcgcagaact
     3961 ctcacagcta ttgctttgag attctggagc ggcggctgta caagtccgga ctcagatctc
     4021 gacaaggtag tggtgctggc tctggtgctg gtagtggcgc tggttccggt gctggctctg
     4081 gcgcgcctcg agcatccaag taccaacaag accaggtggt gaaggaggac agcgtagagg
     4141 ctgtgggcgc ccagctcaag gtctaccacc agcagtacca ggacaagagc cgcgaatatg
     4201 accagctgta tgaagaatac acacggacct cccaggagct gcagatgaag cgcacagcca
     4261 tagaggcctt caacgagacc atcaagatct tcgaagagca gggccagaca caggagaagt
     4321 gcagcaagga gtatttggag cgcttccggc gagagggaaa tgagaaggag atgcagagga
     4381 tcctgctgaa ctccgagcga ctcaagtctc gcatcgcgga gatacacgaa agccgcacga
     4441 agttggagca ggatctgcgg gcgcaggcct ccgacaaccg tgagatcgac aagcgcatga
     4501 acagcctcaa acctgacctc atgcagctgc gcaagatcag ggaccagtac ctcgtgtggc
     4561 tcacccagaa aggtgcccga cagaggaaga tcaacgaatg gctgggaatc aagaacgaga
     4621 ctgaggacca gtattcactg atggaggatg aggacgccct cccccaccac gaggagcgca
     4681 cgtgagaatt ctgcagtcga cggtaccgcg ggcccgggat aagtcaacta acttaagcta
     4741 gcaacggttt ccctctagcg ggatcaattc cgcccccccc ccctaacgtt actggccgaa
     4801 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     4861 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     4921 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     4981 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     5041 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     5101 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     5161 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     5221 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     5281 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga taataccatg
     5341 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5401 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5461 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5521 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5581 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5641 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5701 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5761 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5821 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5881 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatcg
     5941 atgatctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     6001 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6061 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6121 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6181 ggtgggctct atggcttctg aggcggaaag aacctgcagc ccaagcttgg cgtaatcatg
     6241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6301 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tcttaaaagt
     6421 tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat
     6481 aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat
     6541 atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt
     6601 ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaatctag
     6661 tatacgcgtt taattaatcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     6721 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6781 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6841 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6901 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6961 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7021 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7081 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7141 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7201 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7261 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7321 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7381 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7441 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7501 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7561 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7621 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7681 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7741 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7801 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7861 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7921 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7981 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8041 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8101 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8161 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8221 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8281 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8341 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8401 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8461 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8521 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8581 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8641 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8701 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8761 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8821 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
