LOCUS       sequence-262616-       17502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-inactivating DG-Rabies (SiR) expressing FLPo and destabilised 
            mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17502)
  AUTHORS   Ciabatti E, Gonzalez-Rueda A, Mariotti L, Morgese F, Tripodi M
  TITLE     Life-Long Genetic and Functional Access to Neural Circuits Using 
            Self-Inactivating Rabies Virus.
  JOURNAL   Cell. 2017 Jul 1. pii: S0092-8674(17)30697-9. doi: 
            10.1016/j.cell.2017.06.014.
  PUBMED    28689641
REFERENCE   2  (bases 1 to 17502)
  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"; journalName: "Cell. 2017 
            Jul 1. pii: S0092-8674(17)30697-9. doi: 10.1016/j.cell.2017.06.014."
COMMENT     Sequence Label: pSAD-F3-NPEST-FLPo-2A-mCherryPEST
FEATURES             Location/Qualifiers
     source          1..17502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      13..22
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(19..24,25..45,46..1314)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     CDS             1324..1380
                     /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"
     CDS             1387..2091
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1529..1547)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1797..1816
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3968..3979
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9077..9094)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9083..9307
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9353..9781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9440..9459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9650..9671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9790..9810)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9795..10124
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9975..10110
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10037..10056
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10172..10219
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10238..10612
                     /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    10742..10863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10779..10798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10833..10852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10912..10928)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10912..10928)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10925..10947)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10936..10952
                     /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(10960..10966,10967..10984,10985..10990))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11005..11026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11143..11160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11314..11902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11394..11413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12073..12864,12865..12933))
                     /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     12696..12715
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12934..13038)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13113..13132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13304..13683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        13684..13887
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     13838..13858
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     13932..13951
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13932..13950
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             15446..15466
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 ggccgcggta ccgccaccat ggctcctaag aagaagagga aggtgatgag ccagttcgac
       61 atcctgtgca agaccccccc caaggtgctg gtgcggcagt tcgtggagag attcgagagg
      121 cccagcggcg agaagatcgc cagctgtgcc gccgagctga cctacctgtg ctggatgatc
      181 acccacaacg gcaccgccat caagagggcc accttcatga gctacaacac catcatcagc
      241 aacagcctga gcttcgacat cgtgaacaag agcctgcagt tcaagtacaa gacccagaag
      301 gccaccatcc tggaggccag cctgaagaag ctgatccccg cctgggagtt caccatcatc
      361 ccttacaacg gccagaagca ccagagcgac atcaccgaca tcgtgtccag cctgcagctg
      421 cagttcgaga gcagcgagga ggccgacaag ggcaacagcc acagcaagaa gatgctgaag
      481 gccctgctgt ccgagggcga gagcatctgg gagatcaccg agaagatcct gaacagcttc
      541 gagtacacca gcaggttcac caagaccaag accctgtacc agttcctgtt cctggccaca
      601 ttcatcaact gcggcaggtt cagcgacatc aagaacgtgg accccaagag cttcaagctg
      661 gtgcagaaca agtacctggg cgtgatcatt cagtgcctgg tgaccgagac caagacaagc
      721 gtgtccaggc acatctactt tttcagcgcc agaggcagga tcgaccccct ggtgtacctg
      781 gacgagttcc tgaggaacag cgagcccgtg ctgaagagag tgaacaggac cggcaacagc
      841 agcagcaaca agcaggagta ccagctgctg aaggacaacc tggtgcgcag ctacaacaag
      901 gccctgaaga agaacgcccc ctaccccatc ttcgctatca agaacggccc taagagccac
      961 atcggcaggc acctgatgac cagctttctg agcatgaagg gcctgaccga gctgacaaac
     1021 gtggtgggca actggagcga caagagggcc tccgccgtgg ccaggaccac ctacacccac
     1081 cagatcaccg ccatccccga ccactacttc gccctggtgt ccaggtacta cgcctacgac
     1141 cccatcagca aggagatgat cgccctgaag gacgagacca accccatcga ggagtggcag
     1201 cacatcgagc agctgaaggg cagcgccgag ggcagcatca gataccccgc ctggaacggc
     1261 atcatcagcc aggaggtgct ggactacctg agcagctaca tcaacaggcg gatcggatcc
     1321 ggagccacga acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtcct
     1381 accggtgtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1441 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1501 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1561 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1621 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1681 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1741 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1801 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1861 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1921 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1981 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2041 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gggatatctc
     2101 agccatggct tcccgccgga ggtggaggag caggatgatg gcacgctgcc catgtcttgt
     2161 gcccaggaga gcgggatgga ccgtcaccct gcagcctgtg cttctgctag gatcaatgtg
     2221 tgactcgagg gcgcgcctac ccgcggtagc ttttcagtcg agaaaaaaac attagatcag
     2281 aagaacaact ggcaacactt ctcaacctga gacttacttc aagatgctcg atcctggaga
     2341 ggtctatgat gaccctattg acccaatcga gttagaggct gaacccagag gaacccccat
     2401 tgtccccaac atcttgagga actctgacta caatctcaac tctcctttga tagaagatcc
     2461 tgctagacta atgttagaat ggttaaaaac agggaataga ccttatcgga tgactctaac
     2521 agacaattgc tccaggtctt tcagagtttt gaaagattat ttcaagaagg tagatttggg
     2581 ttctctcaag gtgggcggaa tggctgcaca gtcaatgatt tctctctggt tatatggtgc
     2641 ccactctgaa tccaacagga gccggagatg tataacagac ttggcccatt tctattccaa
     2701 gtcgtccccc atagagaagc tgttgaatct cacgctagga aatagagggc tgagaatccc
     2761 cccagaggga gtgttaagtt gccttgagag ggttgattat gataatgcat ttggaaggta
     2821 tcttgccaac acgtattcct cttacttgtt cttccatgta atcaccttat acatgaacgc
     2881 cctagactgg gatgaagaaa agaccatcct agcattatgg aaagatttaa cctcagtgga
     2941 catcgggaag gacttggtaa agttcaaaga ccaaatatgg ggactgctga tcgtgacaaa
     3001 ggactttgtt tactcccaaa gttccaattg tctttttgac agaaactaca cacttatgct
     3061 aaaagatctt ttcttgtctc gcttcaactc cttaatggtc ttgctctctc ccccagagcc
     3121 ccgatactca gatgacttga tatctcaact atgccagctg tacattgctg gggatcaagt
     3181 cttgtctatg tgtggaaact ccggctatga agtcatcaaa atattggagc catatgtcgt
     3241 gaatagttta gtccagagag cagaaaagtt taggcctctc attcattcct tgggagactt
     3301 tcctgtattt ataaaagaca aggtaagtca acttgaagag acgttcggtc cctgtgcaag
     3361 aaggttcttt agggctctgg atcaattcga caacatacat gacttggttt ttgtgtttgg
     3421 ctgttacagg cattgggggc acccatatat agattatcga aagggtctgt caaaactata
     3481 tgatcaggtt caccttaaaa aaatgataga taagtcctac caggagtgct tagcaagcga
     3541 cctagccagg aggatcctta gatggggttt tgataagtac tccaagtggt atctggattc
     3601 aagattccta gcccgagacc accccttgac tccttatatc aaaacccaaa catggccacc
     3661 caaacatatt gtagacttgg tgggggatac atggcacaag ctcccgatca cgcagatctt
     3721 tgagattcct gaatcaatgg atccgtcaga aatattggat gacaaatcac attctttcac
     3781 cagaacgaga ctagcttctt ggctgtcaga aaaccgaggg gggcctgttc ctagcgaaaa
     3841 agttattatc acggccctgt ctaagccgcc tgtcaatccc cgagagtttc tgaggtctat
     3901 agacctcgga ggattgccag atgaagactt gataattggc ctcaagccaa aggaacggga
     3961 attgaagatt gaaggtcgat tctttgctct aatgtcatgg aatctaagat tgtattttgt
     4021 catcactgaa aaactcttgg ccaactacat cttgccactt tttgacgcgc tgactatgac
     4081 agacaacctg aacaaggtgt ttaaaaagct gatcgacagg gtcaccgggc aagggctttt
     4141 ggactattca agggtcacat atgcatttca cctggactat gaaaagtgga acaaccatca
     4201 aagattagag tcaacagagg atgtattttc tgtcctagat caagtgtttg gattgaagag
     4261 agtgttttct agaacacacg agttttttca aaaggcctgg atctattatt cagacagatc
     4321 agacctcatc gggttacggg aggatcaaat atactgctta gatgcgtcca acggcccaac
     4381 ctgttggaat ggccaggatg gcgggctaga aggcttacgg cagaagggct ggagtctagt
     4441 cagcttattg atgatagata gagaatctca aatcaggaac acaagaacca aaatactagc
     4501 tcaaggagac aaccaggttt tatgtccgac atacatgttg tcgccagggc tatctcaaga
     4561 ggggctcctc tatgaattgg agagaatatc aaggaatgca ctttcgatat acagagccgt
     4621 cgaggaaggg gcatctaagc tagggctgat catcaagaaa gaagagacca tgtgtagtta
     4681 tgacttcctc atctatggaa aaaccccttt gtttagaggt aacatattgg tgcctgagtc
     4741 caaaagatgg gccagagtct cttgcgtctc taatgaccaa atagtcaacc tcgccaatat
     4801 aatgtcgaca gtgtccacca atgcgctaac agtggcacaa cactctcaat ctttgatcaa
     4861 accgatgagg gattttctgc tcatgtcagt acaggcagtc tttcactacc tgctatttag
     4921 cccaatctta aagggaagag tttacaagat tctgagcgct gaaggggaga gctttctcct
     4981 agccatgtca aggataatct atctagatcc ttctttggga gggatatctg gaatgtccct
     5041 cggaagattc catatacgac agttctcaga ccctgtctct gaagggttat ccttctggag
     5101 agagatctgg ttaagctccc aagagtcctg gattcacgcg ttgtgtcaag aggctggaaa
     5161 cccagatctt ggagagagaa cactcgagag cttcactcgc cttctagaag atccgaccac
     5221 cttaaatatc agaggagggg ccagtcctac cattctactc aaggatgcaa tcagaaaggc
     5281 tttatatgac gaggtggaca aggtggaaaa ttcagagttt cgagaggcaa tcctgttgtc
     5341 caagacccat agagataatt ttatactctt cttaatatct gttgagcctc tgtttcctcg
     5401 atttctcagt gagctattca gttcgtcttt tttgggaatc cccgagtcaa tcattggatt
     5461 gatacaaaac tcccgaacga taagaaggca gtttagaaag agtctctcaa aaactttaga
     5521 agaatccttc tacaactcag agatccacgg gattagtcgg atgacccaga cacctcagag
     5581 ggttgggggg gtgtggcctt gctcttcaga gagggcagat ctacttaggg agatctcttg
     5641 gggaagaaaa gtggtaggca cgacagttcc tcacccttct gagatgttgg gattacttcc
     5701 caagtcctct atttcttgca cttgtggagc aacaggagga ggcaatccta gagtttctgt
     5761 atcagtactc ccgtcctttg atcagtcatt tttttcacga ggccccctaa agggatactt
     5821 gggctcgtcc acctctatgt cgacccagct attccatgca tgggaaaaag tcactaatgt
     5881 tcatgtggtg aagagagctc tatcgttaaa agaatctata aactggttca ttactagaga
     5941 ttccaacttg gctcaagctc taattaggaa cattatgtct ctgacaggcc ctgatttccc
     6001 tctagaggag gcccctgtct tcaaaaggac ggggtcagcc ttgcataggt tcaagtctgc
     6061 cagatacagc gaaggagggt attcttctgt ctgcccgaac ctcctctctc atatttctgt
     6121 tagtacagac accatgtctg atttgaccca agacgggaag aactacgatt tcatgttcca
     6181 gccattgatg ctttatgcac agacatggac atcagagctg gtacagagag acacaaggct
     6241 aagagactct acgtttcatt ggcacctccg atgcaacagg tgtgtgagac ccattgacga
     6301 cgtgaccctg gagacctctc agatcttcga gtttccggat gtgtcgaaaa gaatatccag
     6361 aatggtttct ggggctgtgc ctcacttcca gaggcttccc gatatccgtc tgagaccagg
     6421 agattttgaa tctctaagcg gtagagaaaa gtctcaccat atcggatcag ctcaggggct
     6481 cttatactca atcttagtgg caattcacga ctcaggatac aatgatggaa ccatcttccc
     6541 tgtcaacata tacggcaagg tttcccctag agactatttg agagggctcg caaggggagt
     6601 attgatagga tcctcgattt gcttcttgac aagaatgaca aatatcaata ttaatagacc
     6661 tcttgaattg gtctcagggg taatctcata tattctcctg aggctagata accatccctc
     6721 cttgtacata atgctcagag aaccgtctct tagaggagag atattttcta tccctcagaa
     6781 aatccccgcc gcttatccaa ccactatgaa agaaggcaac agatcaatct tgtgttatct
     6841 ccaacatgtg ctacgctatg agcgagagat aatcacggcg tctccagaga atgactggct
     6901 atggatcttt tcagacttta gaagtgccaa aatgacgtac ctatccctca ttacttacca
     6961 gtctcatctt ctactccaga gggttgagag aaacctatct aagagtatga gagataacct
     7021 gcgacaattg agttctttga tgaggcaggt gctgggcggg cacggagaag ataccttaga
     7081 gtcagacgac aacattcaac gactgctaaa agactcttta cgaaggacaa gatgggtgga
     7141 tcaagaggtg cgccatgcag ctagaaccat gactggagat tacagcccca acaagaaggt
     7201 gtcccgtaag gtaggatgtt cagaatgggt ctgctctgct caacaggttg cagtctctac
     7261 ctcagcaaac ccggcccctg tctcggagct tgacataagg gccctctcta agaggttcca
     7321 gaaccctttg atctcgggct tgagagtggt tcagtgggca accggtgctc attataagct
     7381 taagcctatt ctagatgatc tcaatgtttt cccatctctc tgccttgtag ttggggacgg
     7441 gtcagggggg atatcaaggg cagtcctcaa catgtttcca gatgccaagc ttgtgttcaa
     7501 cagtctttta gaggtgaatg acctgatggc ttccggaaca catccactgc ctccttcagc
     7561 aatcatgagg ggaggaaatg atatcgtctc cagagtgata gatcttgact caatctggga
     7621 aaaaccgtcc gacttgagaa acttggcaac ctggaaatac ttccagtcag tccaaaagca
     7681 ggtcaacatg tcctatgacc tcattatttg cgatgcagaa gttactgaca ttgcatctat
     7741 caaccggatc accctgttaa tgtccgattt tgcattgtct atagatggac cactctattt
     7801 ggtcttcaaa acttatggga ctatgctagt aaatccaaac tacaaggcta ttcaacacct
     7861 gtcaagagcg ttcccctcgg tcacagggtt tatcacccaa gtaacttcgt ctttttcatc
     7921 tgagctctac ctccgattct ccaaacgagg gaagtttttc agagatgctg agtacttgac
     7981 ctcttccacc cttcgagaaa tgagccttgt gttattcaat tgtagcagcc ccaagagtga
     8041 gatgcagaga gctcgttcct tgaactatca ggatcttgtg agaggatttc ctgaagaaat
     8101 catatcaaat ccttacaatg agatgatcat aactctgatt gacagtgatg tagaatcttt
     8161 tctagtccac aagatggttg atgatcttga gttacagagg ggaactctgt ctaaagtggc
     8221 tatcattata gccatcatga tagttttctc caacagagtc ttcaacgttt ccaaacccct
     8281 aactgacccc tcgttctatc caccgtctga tcccaaaatc ctgaggcact tcaacatatg
     8341 ttgcagtact atgatgtatc tatctactgc tttaggtgac gtccctagct tcgcaagact
     8401 tcacgacctg tataacagac ctataactta ttacttcaga aagcaagtca ttcgagggaa
     8461 cgtttatcta tcttggagtt ggtccaacga cacctcagtg ttcaaaaggg tagcctgtaa
     8521 ttctagcctg agtctgtcat ctcactggat caggttgatt tacaagatag tgaagactac
     8581 cagactcgtt ggcagcatca aggatctatc cagagaagtg gaaagacacc ttcataggta
     8641 caacaggtgg atcaccctag aggatatcag atctagatca tccctactag actacagttg
     8701 cctgtgaacc ggatactcct ggaagcctgc ccatgctaag actcttgtgt gatgtatctt
     8761 gaaaaaaaca agatcctaaa tctgaacctt tggttgtttg attgtttttc tcatttttgt
     8821 tgtttatttg ttaagcgtgg gtcggcatgg catctccacc tcctcgcggt ccgacctggg
     8881 catccgaagg aggacgcacg tccactcgga tggctaaggg agaagggcga attccagcac
     8941 actggcggcc gttactagtg gatccgagct cggtaccgag ctcggatcca ctagtccagt
     9001 gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt
     9061 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     9121 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     9181 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     9241 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     9301 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     9361 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     9421 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     9481 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     9541 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     9601 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     9661 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     9721 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     9781 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     9841 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     9901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     9961 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
    10021 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
    10081 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
    10141 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
    10201 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
    10261 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
    10321 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
    10381 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
    10441 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
    10501 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
    10561 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
    10621 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
    10681 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
    10741 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    10801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    10861 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    10921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    10981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    11041 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    11101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    11161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    11221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    11281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    11341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    11401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    11461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    11521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    11581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    11641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    11701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    11761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    11941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    12001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    12061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    12121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    12181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    12241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    12301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    12361 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    12421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    12481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    12541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    12601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    12661 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    12721 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    12781 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    12841 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    12901 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    12961 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    13021 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
    13081 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    13141 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
    13201 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
    13261 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
    13321 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    13381 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    13441 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    13501 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    13561 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    13621 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
    13681 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
    13741 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
    13801 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
    13861 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
    13921 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagat taagcgtctg
    13981 atgagtccgt gagaacgaaa cccggagtcc cgggtcacgc ttaacaacca gatcaaagaa
    14041 aaaacagaca ttgtcaattg caaagcaaaa atgtaacacc cctacaatgg atgccgacaa
    14101 gattgtattc aaagtcaata atcaggtggt ctctttgaag cctgagatta tcgtggatca
    14161 atatgagtac aagtaccctg ccatcaaaga tttgaaaaag ccctgtataa ccctaggaaa
    14221 ggctcccgat ttaaataaag catacaagtc agttttgtca ggcatgagcg ccgccaaact
    14281 taatcctgac gatgtatgtt cctatttggc agcggcaatg cagttttttg aggggacatg
    14341 tccggaagac tggaccagct atggaattgt gattgcacga aaaggagata agatcacccc
    14401 aggttctctg gtggagataa aacgtactga tgtagaaggg aattgggctc tgacaggagg
    14461 catggaactg acaagagacc ccactgtccc tgagcatgcg tccttagtcg gtcttctctt
    14521 gagtctgtat aggttgagca aaatatccgg gcaaaacact ggtaactata agacaaacat
    14581 tgcagacagg atagagcaga tttttgagac agcccctttt gttaaaatcg tggaacacca
    14641 tactctaatg acaactcaca aaatgtgtgc taattggagt actataccaa acttcagatt
    14701 tttggccgga acctatgaca tgtttttctc ccggattgag catctatatt cagcaatcag
    14761 agtgggcaca gttgtcactg cttatgaaga ctgttcagga ctggtatcat ttactgggtt
    14821 cataaaacaa atcaatctca ccgctagaga ggcaatacta tatttcttcc acaagaactt
    14881 tgaggaagag ataagaagaa tgtttgagcc agggcaggag acagctgttc ctcactctta
    14941 tttcatccac ttccgttcac taggcttgag tgggaaatct ccttattcat caaatgctgt
    15001 tggtcacgtg ttcaatctca ttcactttgt aggatgctat atgggtcaag tcagatccct
    15061 aaatgcaacg gttattgctg catgtgctcc tcatgaaatg tctgttctag ggggctatct
    15121 gggagaggaa ttcttcggga aagggacatt tgaaagaaga ttcttcagag atgagaaaga
    15181 acttcaagaa tacgaggcgg ctgaactgac aaagactgac gtagcactgg cagatgatgg
    15241 aactgtcaac tctgacgacg aggactactt ttcaggtgaa accagaagtc cggaggctgt
    15301 ttatactcga atcatgatga atggaggtcg actaaagaga tctcacatac ggagatatgt
    15361 ctcagtcagt tccaatcatc aagcccgtcc aaactcattc gccgagtttc taaacaagac
    15421 atattcgagt gactcaggtt ccggagagaa cctctacttc caatcgggat ccggtagcca
    15481 tggcttcccg ccggaggtgg aggagcagga tgatggcacg ctgcccatgt cttgtgccca
    15541 ggagagcggg atggaccgtc accctgcagc ctgtgcttct gctaggatca atgtgtaaga
    15601 agttgaataa caaaatgccg gaaatctacg gattgtgtat atccatcatg aaaaaaacta
    15661 acacccctcc tttcgaacca tcccaaacat gagcaagatc tttgtcaatc ctagtgctat
    15721 tagagccggt ctggccgatc ttgagatggc tgaagaaact gttgatctga tcaatagaaa
    15781 tatcgaagac aatcaggctc atctccaagg ggaacccata gaggtggaca atctccctga
    15841 ggatatgggg cgacttcacc tggatgatgg aaaatcgccc aaccatggtg agatagccaa
    15901 ggtgggagaa ggcaagtatc gagaggactt tcagatggat gaaggagagg atcctagctt
    15961 cctgttccag tcatacctgg aaaatgttgg agtccaaata gtcagacaaa tgaggtcagg
    16021 agagagattt ctcaagatat ggtcacagac cgtagaagag attatatcct atgtcgcggt
    16081 caactttccc aaccctccag gaaagtcttc agaggataaa tcaacccaga ctactggccg
    16141 agagctcaag aaggagacaa cacccactcc ttctcagaga gaaagccaat catcgaaagc
    16201 caggatggcg gctcaaattg cttctggccc tccagccctt gaatggtcgg ctaccaatga
    16261 agaggatgat ctatcagtgg aggctgagat cgctcaccag attgcagaaa gtttctccaa
    16321 aaaatataag tttccctctc gatcctcagg gatactcttg tataattttg agcaattgaa
    16381 aatgaacctt gatgatatag ttaaagaggc aaaaaatgta ccaggtgtga cccgtttagc
    16441 ccatgacggg tccaaactcc ccctaagatg tgtactggga tgggtcgctt tggccaactc
    16501 taagaaattc cagttgttag tcgaatccga caagctgagt aaaatcatgc aagatgactt
    16561 gaatcgctat acatcttgct aaccgaacct ctcccctcag tccctctaga caataaaatc
    16621 cgagatgtcc caaagtcaac atgaaaaaaa caggcaacac cactgataaa tcgatgaacc
    16681 tcctacgtaa gatagtgaaa aaccgcaggg acgaggacac tcaaaaatcc tctcccgcgt
    16741 cagcccctct ggatgacgat gacttgtggc ttccaccccc tgaatacgtc ccgctgaaag
    16801 aacttacagg caagaagaac atgaggaact tttgtatcaa cggaagggtt aaagtgtgta
    16861 gcccgaatgg ttactcgttc aggatcctgc ggcacattct gaaatcattc gacgagatat
    16921 attctgggaa tcataggatg atcgggttag tcaaagtggt tattggactg gctttgtcag
    16981 gatctccagt ccctgagggc ctgaactggg tatacaaatt gaggagaacc tttatcttcc
    17041 agtgggctga ttccaggggc cctcttgaag gggaggagtt ggaatactct caggagatca
    17101 cttgggatga tgatactgag ttcgtcggat tgcaaataag agtgattgca aaacagtgtc
    17161 atatccaggg cagagtctgg tgtatcaaca tgaacccgag agcatgtcaa ctatggtctg
    17221 acatgtctct tcagacacaa aggtccgaag aggacaaaga ttcctctctg cttctagaat
    17281 aacgtacgtc agattatatc ccgcaaattt atcacttgtt tacctctgga ggagagaaca
    17341 tatgggctca actccaaccc ttgggagcaa tataacaaaa aacatgttat ggtgccatta
    17401 aaccgctgca tttcatcaaa gtcaagttga ttacctttac attttgatcc tcttggatgt
    17461 gaaaaaaact attaacatcc ctcaaaggac ctgcaggtac gc
//
