LOCUS       sequence-455547-       15176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of beta actin driven BFP with multiple cloning site at 3'
            end. Also contains heat-shock activated mCherry reporter and 
            CreERT2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15176)
  AUTHORS   Siniscalco AM, Perera RP, Greenslade JE, Veeravenkatasubramanian H, 
            Masters A, Doll HM, Raj B
  TITLE     Barcoding Notch signaling in the developing brain.
  JOURNAL   Development. 2024 Nov 22:dev.203102. doi: 10.1242/dev.203102.
  PUBMED    39575683
REFERENCE   2  (bases 1 to 15176)
  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: 
            "Development. 2024 Nov 22:dev.203102. doi: 10.1242/dev.203102."
COMMENT     Sequence Label: p133Tol2-Olactb:MCS, hsp70l-mcherry-CreERT2
FEATURES             Location/Qualifiers
     source          1..15176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(73..966)
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     CDS             complement(79..1014)
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             complement(1027..2052)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             complement(2059..2112)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(2113..2820)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2388..2407)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2657..2675
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2817..2826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2847..2871)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(4764..4951,4952..4963)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4997..5013)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4997..5013)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5050..5070)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5050..5068)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5089..5105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5089..5105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5102..5124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5113..5129
                     /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(5137..5143,5144..5161,5162..5167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5182..5203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5320..5337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5491..6079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5571..5590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6250..7041,7042..7110))
                     /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     6873..6892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7111..7215)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7241..7696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7378..7399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7590..7609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7823..7845
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7837..7854
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7838..7854
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7864..7882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(9734..9745,9746..9908)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             join(14379..14381,14382..14384,14385..15098)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(14685..14704)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     15032..15053
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tacgtaatac gactcactat agttctagag gctcgagagg ccttgaattc cgccagggag
       61 ctcagatctt catcaagctg tggcagggaa accctctgcc tcccccgtga tgtaatactt
      121 ttgcaaggaa tgcgatgaag tagagcccgc agtggccaag tggctttggt ccgtctcctc
      181 cacggatgcc cctccacggc tagtgggcgc atgtaggcgg tgggcgtccg ccgcctccag
      241 cagcaggtca tagaggggca ccacgttctt gcacttcatg ctgtacagat gctccatgcc
      301 tttgttactc atgtgcctga tgtgggagag gatgaggagg agctgggcca gccgctggtg
      361 ctgctgctgc agggtcaggc ctgccttggc catcaggtgg atcaaagtgt ctgtgatctt
      421 gtccaggact cggtggatat ggtccttctc ttccagagac ttcagggtgc tggacagaaa
      481 tgtgtacact ccagaattaa gcaaaataat agatttgagg cacacaaact cctctccctg
      541 cagattcatc atgcggaacc gagatgatgt agccagcagc atgtcgaaga tctccaccat
      601 gccctctaca cattttccct ggttcctgtc caagagcaag ttaggagcaa acagtagctt
      661 cactgggtgc tccatggagc gccagacgag accaatcatc aggatctcta gccaggcaca
      721 ttctagaagg tggacctgat catggagggt caaatccaca aagcctggca ccctcttcgc
      781 ccagttgatc atgtgaacca gctccctgtc tgccaggttg gtcagtaagc ccatcatcga
      841 agcttcactg aagggtctgg taggatcata ctcggaatag agtatggggg gctcagcatc
      901 caacaaggca ctgaccatct ggtcggccgt cagggacaag gccaggctgt tcttcttaga
      961 gcgtttgatc atgagcgggc ttggccaaag gttggcagct ctcatgtctc cagcagatgg
     1021 ctcgagatcg ccatcttcca gcaggcgcac cattgcccct gtttcactat ccaggttacg
     1081 gatatagttc atgacaatat ttacattggt ccagccacca gcttgcatga tctccggtat
     1141 tgaaactcca gcgcgggcca tatctcgcgc ggctccgaca cgggcactgt gtccagacca
     1201 ggccaggtat ctctgaccag agtcatcctt agcgccgtaa atcaatcgat gagttgcttc
     1261 aaaaatccct tccagggcgc gagttgatag ctggctggtg gcagatggcg cggcaacacc
     1321 attttttctg acccggcaaa acaggtagtt attcggatca tcagctacac cagagacgga
     1381 aatccatcgc tcgaccagtt tagttacccc caggctaagt gccttctcta cacctgcggt
     1441 gctaaccagc gttttcgttc tgccaatatg gattaacatt ctcccaccgt cagtacgtga
     1501 gatatcttta accctgatcc tggcaatttc ggctatacgt aacagggtgt tataagcaat
     1561 ccccagaaat gccagattac gtatatcctg gcagcgatcg ctattttcca tgagtgaacg
     1621 aacctggtcg aaatcagtgc gttcgaacgc tagagcctgt tttgcacgtt caccggcatc
     1681 aacgttttct tttcggatcc gccgcataac cagtgaaaca gcattgctgt cacttggtcg
     1741 tggcagcccg gaccgacgat gaagcatgtt tagctggccc aaatgttgct ggatagtttt
     1801 tactgccaga ccgcgcgcct gaagatatag aagataatcg cgaacatctt caggttctgc
     1861 gggaaaccat ttccggttat tcaacttgca ccatgccgcc cacgaccggc aaacggacag
     1921 aagcattttc caggtatgct cagaaaacgc ctggcgatcc ctgaacatgt ccatcaggtt
     1981 cttgcgaacc tcatcactcg ttgcatcgac cggtaatgca ggcaaatttt ggtgtacggt
     2041 cagtaaattg gagctagcag ggccgggatt ctcctccacg tcaccgcatg ttagaagact
     2101 tcctctgccc tccttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg
     2161 ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt
     2221 gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc
     2281 agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc
     2341 gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat
     2401 tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc
     2461 gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac
     2521 gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg
     2581 gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa
     2641 gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg
     2701 gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgtgcac
     2761 cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct tgctcaccat
     2821 ggtggccggc ccatatggcc cgggccgttc aacttttttg tacaaacttg gcaggaaaaa
     2881 aaaacaatta gaattaattt tatatttata caatttatgg tgcaattgtt tcattatgaa
     2941 aggatttttc tagtcaacaa gtgaaattct cttaataacc aagttattat tagttgtaca
     3001 agtaaaacgc taattttctt ttgtagacaa cattaacaca aattcattta ggacatttgc
     3061 tagtatttat ggttattatg ctcatataaa ataatgtcag caacataaag taaatgcccg
     3121 tctgttcatt gcttttttta aaaatcctta ttgtgaaaac gggaaatata attttctagt
     3181 agacctaaaa taagatgaac ataggcagtc cacatgtaaa aagattacat atacaaagtt
     3241 gattttttga cattaagtac atataatctc tatataattc tgattagcca atagagtaca
     3301 atatattgag acttacctta cagtagattc ttaaatagag cgaataaaca cctcgtcggg
     3361 gaaaaagtcc tgtcaggctg ctcttttctt ctttgtcaag tcgcttctct tcggttgagt
     3421 ttctctgcgg tctctctttc gccctttaaa tacagcagag cgcgcacccg ttcagactct
     3481 acactgttct ggaagcctcc agtgaattcg accaatcaaa acacagcagg cagccgcgtc
     3541 acttcttcag ctgtcattga gaggagccgc tgagaggaat gtagtggaga tttccagagt
     3601 tttaccgtga aattacgcga aacagcggtc aaaaatacaa agagatttaa aatgtttttt
     3661 ttatttgagt gttttattgt acttattgct gaaatatgtg cggttgtgtg atgtgttgtt
     3721 tttccagttg caatgaatga acaaactaaa atgataaaat gtttgaacga ttcagtaaaa
     3781 aaaaatgcaa taaaaaaatc ttgctgaaat gttttacaca gtagcaaaat cgggcctgtg
     3841 gtgttttcag ctacatagaa ggcaaacctg ttttatataa gctaatagct ttggtcagat
     3901 ttgtttaacg tttcaacaaa acaaatggat gtttactgta ctatattgat tggttaaaac
     3961 caatacttta ttctgggatt gcagtcttaa ctatttttgt agtgttaaaa ccaacttgct
     4021 aacttgagag gtcatgttta ccctcagaga aataggcaca agtctctttg tttaatattg
     4081 gagatcatcc tccttgttca gtcgtggtct gtatttaatt ttaatgtata tgattgccgt
     4141 aaaagtgtta gaaagtttac cctggtgcta tgaaatcaat ctgctgcttt tgacccagtt
     4201 agtgtgtctt taatctaact taatgagacc ccagtgtggt caacaatcaa tagaaacaaa
     4261 gtgtgttgtt ttataactgt taactcctat tattttttat cttctgtagc atatagacaa
     4321 aacatgctcg ttctaatagt ttaataaaat taatttgatt tttggaaata accaagcgac
     4381 accccatccg gagtcgacgg gcccatctgg cctgtgtttc agacaccagg gagtcctagg
     4441 agatctgcga agatacggcc acgggtgctc ttgatcctgt ggctgatttt ggactgtgct
     4501 gctcgcagct gctgatgaat cacatacttc ctccattttc ttccactgat tgactgttat
     4561 aatttcccta atttccaggt caaggtgctg tgcattgtgg taatagatgt gacatgacgt
     4621 cacttccaaa ggaccaatga acatgtctga ccaatttcat ataatgtgaa aacgattttc
     4681 ataggcagaa taaataacat ttaaattaaa ctgggcatca gcgcaattca attggtttgg
     4741 taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa
     4801 ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt
     4861 aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa
     4921 gtacagtaat caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc
     4981 ttcagcaagc ccagcagatc cactagttct agagcggccg ccaccgcggt ggagctccag
     5041 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     5101 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5161 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5281 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5341 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5401 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6001 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6061 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     7261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     7381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     7501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     7621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7681 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     7741 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     7801 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     7861 gcgtaatacg actcactata gggcgaattg ggtaccaaat agtaggaatt acccacctgt
     7921 acaagtgctg aaaacttgga tgaataagcc cgtttgccat tttctactgc tattttaaat
     7981 cttttctgta tttgtctgca tttgtcttta cccttgaata aatgttttag ttgttttttt
     8041 tcaattatga ctgtgtttaa cagacattat tacaataata tgtaatagta gtacacacta
     8101 ttattatgta atagtacttg ttgactgtat ttgagaactg gaccgagtga gtgttacgtc
     8161 acccattcaa aatgacttac ttctggctcc aacaaaatga agttaattca gttgccattt
     8221 ttcactgtat ggacatcgcc gtgttggagc tagacgttgc caataagcaa gaaagagccg
     8281 agatgcgtcg agtctgagtc accgttccta tggcaacccc tctaaccaat cagaagtaag
     8341 cttgttggaa gtccacagcc taccacttga aagcgggctg cacaaaatct gtcaaacgtt
     8401 ttgaacgttg gatgtgagag cacatacttt tattaaggca tctggttggt cagtttataa
     8461 tatcaacaac ttgggctaca gaaaagaaaa gttattacag aaattatgat taacaagtac
     8521 atgttaaata aagattttaa tatgaatgcc accactggag cattcatgcc atttggagct
     8581 tcttcctgtt tggatcacta gaaggaggag gtcactcatt acagttctca tatacagtcg
     8641 ttggttggtt ggttggttgg tagattgatt gattgattga ttgattgatt gattgattga
     8701 ttgattgatt gattgattga ttgattgatt gattgattgg tagtcaaaat aagaaataat
     8761 ttccacagat tcattacaga aatgattaaa tgcatacata aaaaactggg gggggggggg
     8821 gatacaacaa cacacttaag tcacatttgc ctacgtaaag aaaagtaaag aaaatcaata
     8881 gctatatttt acatctcctt tttttgctgt cttttaatta gccttgtttt gctgttatct
     8941 tgattgaaac tgtaacttct tcacctgctt cttttctttg taggttttgc cagcccagaa
     9001 acaggcatgg ctgtgcaagg ccagagcacc atgcacaatg cgctgcatgt cttcatgaac
     9061 ggctcaatgt cctcagtcca gggctcagcc aacgacccca tcttcctcct tcaccatgct
     9121 ttcattgaca ggtaacaaac acgtcatgac attagactgc acagtttttg acaaagttca
     9181 tacaatctgt tgtttatagc tgctacaatt agtgaagttt gtgaatgtac ttggatgagc
     9241 agcgaaagat caattgagat caattgttag agtttggttg ccctgcagag caaagaacaa
     9301 aaaataatct ggtggcttta ctgcgtgagg ttattattgg tggaatagaa acacaaaaca
     9361 taattgcatt tatttgttta attttttatc ttatcttaac tttcatcttg catatttgtt
     9421 tcttacatca tttctagcat ctttgagcgc tggctaagaa ctcatcagcc tccccggtcc
     9481 atctacccac gtaccaatgc accaattggc cacaatgacg gctactacat ggtgccattc
     9541 cttcctcttt ataggaatgg agactacctc ctgtccaaca atgctcttgg atacgagtac
     9601 gcctacctgt tggacccagg tcattgcaca acaccagaaa tgccctctga tctgcaaaag
     9661 acgtgaatat ctgttcagac acccatatcc actctgttcc acacaggtca gaggtttgtc
     9721 caggagttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa
     9781 gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa tcttacttga
     9841 gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attaaggaag taaaagtaaa
     9901 agcaagaaag aaaactagag attcttgttt aagcttttaa tctcaaaaaa cattaaatga
     9961 aatgcataca aggttttatc ctgctttaga actgtttgta tttaattatc aaactataag
    10021 acagacaatc taatgccagt acacgctact caaagttgta aaacctcaga tttaacttca
    10081 gtagaagctg attctcaaaa ttgttagtgt caagcctagc tcttttgggg ctgaaaagca
    10141 atcctgcagt gctgaaaagc ctctcacagg cagccgatgc gggaagaggt gtattagtct
    10201 tgatagagag gctgcaaata gcaggaaacg tgagcagaga ctccctggtg tctgaaacac
    10261 aggccagatg ggccctcgag cagggggatc atctaatcaa gcacaaataa ggggcgtgta
    10321 acacaaaagc cagcgaccct ttccaatgca aatcaaactt gcaattcttt gccgttttta
    10381 tcatttaagt gtcggcttaa ggtccactat cagatgtaaa cagccttatc taacaaaggt
    10441 atcattacat tctgaaattc tcaggcatgc aagctagctt atgacgcact agggagtgcc
    10501 acccttcctt tcgccctaac ttcgtgataa ctcgcgcgtt tcactcaaca gctgcatccg
    10561 ccctagtgct actgggagtt gtagtataca agacgcttac aggctgaatg ttctgtcaag
    10621 accccgcctc tagcactttg ggaattcaat tacagtggtc atgaatatta tatctaacca
    10681 cagagctaaa gaaaaaacat accaagacag ttttttttct gttgatgggt ggtcattcta
    10741 cattgaagtg aatggggtct gactggtttg ttttacgtgt cagctagtgc tttcttggtg
    10801 aactgatctg gctcattttt ttactctgac ctcatctgag aattgttcaa aacaaatgca
    10861 cgatatttaa tactgctgat aagaaggatt cgcttcactt gcaggtgaac cctggtgtgg
    10921 caatagtgtt aatgaaaaca aactactcgg cagtaaaagt atgtgacatg aataaaaaaa
    10981 aatcacaaac tggaagaaga tcagatcatg aaagaattgt aagaatgtcc cattgtacga
    11041 tgtgttgtgg taaagtggtg aagtcactta tcaatccatg atggtcagag aactagcagg
    11101 actggttttc ttttctttgt ttttttttat ttccttcatc ctggtactgc ccacttctgg
    11161 ccttcaaaca aagtcaagta ttttttttat tcaaaagcag atcaaattta gaaggtaaca
    11221 tcatctgccc atttgaatca agtctgtaga actatcttag tatgaacttg ccataatgct
    11281 tcataagaag tgattgcaaa tcacctacca cacagctgga tgagctggaa ctctgacaca
    11341 tacattcatt cacctgcaat cacgtaagga gcctactacg aagggacact cctgtgaggc
    11401 aggattacat cctttcccag ccaacccctg tctgtgtttt tccagtttca gcagtcagtc
    11461 cttccagtgt gctcttaact ttaccagaac ccgccctccc agaatactga caaatagaaa
    11521 taaaaatgtg ttttagatgg acgcaccttg aatgggttac agatttcatt tcaggcctgt
    11581 aaataagtaa atgcagtgag ttggcctttg atcgacaaga acgattcatt ccatatgaga
    11641 gtgtttttta tgagagattt aaaactctta cataaagaat tatttacata caactctcga
    11701 gagagaactg tttcacccca caagcagaag cttttatttc agcgggttaa taaaacaaaa
    11761 caaaaatgga catttaaaag ttaggtctgc tgatcagaaa tgcaagaatc tgctaattca
    11821 gaaggaaatg aagcatgttt tgtttgaaac ttgattaaaa agctgaaacg tttccatgta
    11881 taaagtggaa atgtgttgat gaaaataaag accatttaaa acttaaagcc acgaatgaat
    11941 ttaaggaaga ggagatggca tttagcataa aagaattggt gtaactgaga gcgaggaaga
    12001 ggaagtggta caagaggaag aaaagtagtt gctgcttgtg tctcgcagac agtgacatgt
    12061 ttaaatggcc tctgggttat ttaacttgta tgggcagcta tgtttctgga gcaaaaaaag
    12121 tttaaataaa ctgtgaaact aatgaatgga agacaagtaa ggtaaaagca aagaacccta
    12181 agcctataaa acccaaagtc aggaaagtta caccctttgc agtaaaccac agagaggaaa
    12241 taagagcatg tgtgacttta tttgtctctc tctgagttta atcgattaag gtgaatgtat
    12301 agtagcgtat gggttgggtc tgacagtggt tattttcact caataggtca tttatgacta
    12361 aaagaagatg caagtcagcc ttaatgtcct cttatgtgga ccttttttct tcatttgtag
    12421 gggaaagagg aagtagcaag agctctgaga agttaacaag aatccagact aatgccatac
    12481 atggtcaatt ggtattagtg agattgagtt tttaccattg cttagtggga aaagtggtta
    12541 ttgtccatcc tctgacaggg cacttttaca gtccatagtg tcttttgtga gtcatgtgtt
    12601 tggcttggag ggtctgtgac tccatctgag gtggcattct gctttccaga gtgtacagca
    12661 acatccacgt tgtaattgtc ctcaaattgc atcaaacata tcttgcattt ccagactgaa
    12721 atcttgagag acaggtttta agaggaaaag tattcacagg gaggagcagc ctgtgctctg
    12781 ctgtttgaat gatcagataa gcagggggac ttgctcgaca gctttataca ggttcatatt
    12841 ttatatttat ttgaaacttt gtaaactcta tatgtattaa tatttgttca agaaacatta
    12901 tatccttaga ttaggcctca catcaatccc tatcctttta ttttccccta gcccgttact
    12961 gttatcgcct ccctctttct acgctcagtg caccacagcg tgcacccgac gtgccccagt
    13021 gagtgacgct ggaccaatca catgccgcga ttccgaaagt ttacctttta tggaaagagc
    13081 cgggcaacgg acggactata aataccacgt gcccacggct agcaaattca ctctgagcgc
    13141 cgtcacacac agcttgtgcg ggatatcatt cgcctgaaac cgattccctt gaaacgaaaa
    13201 gccccccacc caaaggtaag atagagacga tatttggttt aattcaaatg ttttggaaac
    13261 cttacgattg tttaattgtg cagtttaatt cttaattgga ggtgaccatt agcagaattt
    13321 tgtggccact aaatttattt ttaggcctga aatggttaaa cttttaatta tctttttaaa
    13381 attgacaatt ttgtttgttt tggcatcaat caagttgtac caaaattagc cacttaaacg
    13441 cacaggtgcc aattgattgt aaccggcaaa tggcttttca gtttggtttt aattgtatat
    13501 tttatatagg caactaaata attactattc gtatcttaca tggtggggtg aatttttcga
    13561 gtccttctac ggacgttacc cgtccgagtt gagggtgtct ggatcctggc gggtcccttt
    13621 gtgcgccgtt acggggtgtg acctacttta gcatattagc ctagcttcat gctaacacgc
    13681 ccttctagac ttgactctga atcctcacat gtaggggggt gggactgtta tacatttgtt
    13741 tgtataaaat gtaataaccc ttaaagcatt ttaaatgtat tcgattaaat tgaccatgta
    13801 actgtagttt gacggcgaag aaaagggcaa atgacaaatc attcattcaa gaaaaggcgt
    13861 ggctttcagg aaatgattga tcagtcatgg gctcgcacac gttttaattt cctactaaag
    13921 cttgacgggt cgtggtgctt tatgccagca acgacttcgc acaaattatg acgcagtgat
    13981 taagaaattt ttcagtcttg tgtaacaatg ggagggaacg gggacaaagt agagctgtaa
    14041 tgaaccagtg ggagtaggct ttcgtcgccg ggtccgatag tctaaagtcc ctcccctctt
    14101 gctgtacttt ggccgcgtgc cttgccggta gccgcaaagc tgctcaaaac cgaactatgt
    14161 ccttatatgg taacaagaga atgcagcacc acttcctttt gtctgggctc ggaatgtagc
    14221 gctctacctc caccaagcgg acactttggg gaactgccgc tccctgacag gaaatgggac
    14281 atctgactgc aaagcagatg cagatgggcg tgtctttggt gttaacgcct tttttgtttg
    14341 ttcttttcca gtttagccgt cgacaccggt ctgacaacat ggtgagcaag ggcgaggagc
    14401 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
    14461 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
    14521 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg
    14581 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
    14641 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
    14701 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
    14761 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacgccatca
    14821 gcgacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
    14881 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
    14941 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
    15001 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
    15061 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggcgtacga cctcgagaca
    15121 aatggcagcc cgggggatcc aaactcgaga aagctagcaa accatggtac gatgat
//
