LOCUS       sequence-212111-       11948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  H2BtdTomato_p2A_HygroR_p2A_CreERt2 targeting plasmid with cloning 
            sites ready for homology arms.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11948)
  AUTHORS   Hartman BH, Bscke R, Ellwanger DC, Keymeulen S, Scheibinger M, 
            Heller S
  TITLE     Fbxo2(VHC) mouse and embryonic stem cell reporter lines delineate in
            vitro-generated inner ear sensory epithelia cells and enable otic 
            lineage selection and Cre-recombination.
  JOURNAL   Dev Biol. 2018 Sep 1. pii: S0012-1606(18)30499-8. doi: 
            10.1016/j.ydbio.2018.08.013.
  PUBMED    30179592
REFERENCE   2  (bases 1 to 11948)
  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: "Dev Biol. 
            2018 Sep 1. pii: S0012-1606(18)30499-8. doi: 
            10.1016/j.ydbio.2018.08.013."
COMMENT     Sequence Label: ptdTHC_PGKneoLox2DTA.2
FEATURES             Location/Qualifiers
     source          1..11948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward 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"
     CDS             734..1111
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             1118..2545
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2555..2611
                     /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             2612..3634
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2612..3634
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3647..3703
                     /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             3704..4732
                     /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="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             4745..5680
                     /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             4793..5686
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     primer_bind     complement(5774..5793)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5839..5894
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5893..5912)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6255..6271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6279..6295
                     /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)."
     primer_bind     6318..6334
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6318..6334
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    6348..6381
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6411..6910
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             6924..7727
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6987..7006)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7597..7616
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7765..7989
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    8048..8081
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        8160..8659
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             8686..9258
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          9335..9400
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    9474..9698
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9760..9780)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9760..9778)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9799..9815)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9823..9839
                     /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(9847..9853,9854..9871,9872..9877))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9892..9913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10030..10047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10201..10789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10281..10300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10960..11751,11752..11820))
                     /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     11583..11602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11821..11925)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgcctag gagatctacg cgtcaattga tgcatcccgg gtttattcat
      721 gaggccggcc accatgccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa
      781 gaaggcggtg actaaggcgc agaagaaaga cggcaagaag cgcaagcgca gccgcaagga
      841 gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca ccggcatttc
      901 gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg
      961 tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca
     1021 gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac
     1081 taaggccgtc accaagtaca ccagcgctaa atccaccatg gtgagcaagg gagaggaggt
     1141 catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt
     1201 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     1261 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta
     1321 cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt
     1381 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt
     1441 gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac
     1501 caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac
     1561 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct
     1621 gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt
     1681 gcaactgccc ggctactact acgtggacac caagctggac atcacctccc acaacgagga
     1741 ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctggggca
     1801 tggcaccggc agcaccggca gcggcagctc cggcaccgcc tcctccgagg acaacaacat
     1861 ggccgtcatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca
     1921 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1981 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt
     2041 catgtacggc tccaaggcgt acgtgaagca ccccgccgac atccccgatt acaagaagct
     2101 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt
     2161 gaccgtgacc caggactcct ccctgcagga cggcacgctg atctacaagg tgaagatgcg
     2221 cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2281 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct
     2341 gaagctgaag gacggcggcc gctacctggt ggagttcaag accatctaca tggccaagaa
     2401 gcccgtgcaa ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa
     2461 cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct
     2521 gtacggcatg gacgagctgt acaagggaag cggagctact aacttcagcc tgctgaagca
     2581 ggctggagac gtggaggaga accctggacc tatgaaaaag cctgaactca ccgcgacgtc
     2641 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     2701 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     2761 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     2821 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
     2881 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     2941 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     3001 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3061 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3121 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3181 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     3241 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     3301 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     3361 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     3421 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     3481 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     3541 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     3601 ccgacgcccc agcactcgtc cgagggcaaa ggaacgcggc tccggcgcaa caaatttctc
     3661 tctcctcaaa caagcaggcg atgttgaaga aaatcccggg cccatgtcca atttactgac
     3721 cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa
     3781 cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct
     3841 gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc
     3901 agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa
     3961 aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg
     4021 accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga
     4081 tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg
     4141 ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat
     4201 tgcttataac accctgttac gtatagccga aattgccagg atcagggtta aagatatctc
     4261 acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac
     4321 cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc
     4381 cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg
     4441 tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga
     4501 agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg
     4561 gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat
     4621 accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg
     4681 taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg atctcgagcc
     4741 atctgctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc
     4801 taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga
     4861 tgctgagccc cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc
     4921 gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc
     4981 gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg
     5041 tgcctggcta gagatcctga tgattggtct cgtctggcgc tccatggagc acccagtgaa
     5101 gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat
     5161 ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca
     5221 gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt
     5281 tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa
     5341 gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca
     5401 ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg
     5461 catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct
     5521 ggaggcggcg gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga
     5581 ggagacggac caaagccact tggccactgc gggctctact tcatcgcatt ccttgcaaaa
     5641 gtattacatc acgggggagg cagagggttt ccctgccaca gcttgatagc ggccgcactc
     5701 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     5761 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     5821 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     5881 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     5941 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     6001 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     6061 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     6121 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     6181 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
     6241 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6301 cacaaaaact aggccgctct agaactagtg gatccggaac ccttaatata acttcgtata
     6361 atgtatgcta tacgaagtta ttaggtccct cgacctgcag gaattctacc gggtagggga
     6421 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     6481 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     6541 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     6601 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     6661 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     6721 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     6781 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     6841 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     6901 gggcctttcg acctgcagcc aatatgggat cggccattga acaagatgga ttgcacgcag
     6961 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     7021 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     7081 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     7141 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     7201 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     7261 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     7321 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     7381 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     7441 tgttcgccag gctcaaggcg cgcatgcccg acggcgatga tctcgtcgtg acccatggcg
     7501 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     7561 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     7621 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     7681 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaggg gatcaattct
     7741 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     7801 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     7861 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     7921 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     7981 gctctatggc ttctgaggcg gaaagaacca gctggggctc gaatcaagct gatccggaac
     8041 ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtccct cgacctgcag
     8101 cccaagctag cttatcgata ccgtcgacgg tatcgataag cttgatatcg aattctaccg
     8161 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     8221 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     8281 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     8341 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     8401 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     8461 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     8521 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     8581 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     8641 ctcatctccg ggcctttcga cctgcaggtc ctcgccatgg atcctgatga tgttgttgat
     8701 tcttctaaat cttttgtgat ggaaaacttt tcttcgtacc acgggactaa acctggttat
     8761 gtagattcca ttcaaaaagg tatacaaaag ccaaaatctg gtacacaagg aaattatgac
     8821 gatgattgga aagggtttta tagtaccgac aataaatacg acgctgcggg atactctgta
     8881 gataatgaaa acccgctctc tggaaaagct ggaggcgtgg tcaaagtgac gtatccagga
     8941 ctgacgaagg ttctcgcact aaaagtggat aatgccgaaa ctattaagaa agagttaggt
     9001 ttaagtctca ctgaaccgtt gatggagcaa gtcggaacgg aagagtttat caaaaggttc
     9061 ggtgatggtg cttcgcgtgt agtgctcagc cttcccttcg ctgaggggag ttctagcgtt
     9121 gaatatatta ataactggga acaggcgaaa gcgttaagcg tagaacttga gattaatttt
     9181 gaaacccgtg gaaaacgtgg ccaagatgcg atgtatgagt atatggctca agcctgtgca
     9241 ggaaatcgtg tcaggcgatc tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     9301 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     9361 atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct atggaactga
     9421 tgaatgggag cagtggtgga atgcagatcc tagagctcgc tgatcagcct cgactgtgcc
     9481 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     9541 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     9601 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     9661 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     9721 ctggggctcg agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     9781 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     9841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     9901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     9961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    10021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    10081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    10141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    10201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    10261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    10321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    10381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    10441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    10501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    10561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    10621 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    10681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    10741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    10801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    10861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    10921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    10981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    11041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    11101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    11161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    11221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    11281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    11341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    11401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    11461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    11521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    11581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    11641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    11701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    11761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    11821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    11881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    11941 agtgccac
//
