LOCUS       sequence-463303-       17682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17682)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 17682)
  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"
COMMENT     Sequence Label: pPMS-985
FEATURES             Location/Qualifiers
     source          1..17682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1092..1316
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     intron          1390..1455
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(1532..2104)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     promoter        complement(2131..2630)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     misc_feature    2657..6997
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     protein_bind    complement(7034..7067)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    7122..7346
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(7384..8187)
                     /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(7495..7514)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8105..8124
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(8206..8253)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(8265..8764)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     protein_bind    complement(8828..8861)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    8931..9155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(9181..9207)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(9214..9240)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(9247..9273)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(9283..9993,9994..9996))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(9325..9346)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9674..9693
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9933..9954
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(10081..10458)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             complement(10477..10530)
                     /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(10546..10566)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(10576..10605)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(10612..10641)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(10648..10677)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(10684..12111)
                     /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
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     regulatory      12138..12147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    12155..12188
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    12212..12346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    12449..12583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    12686..12820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    12949..12982
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(13057..13076)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          13084..14100
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(13138..13160)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(14101..14379)
                     /label=chicken beta-actin promoter
     enhancer        14381..14684
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    14778..15861
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     primer_bind     complement(15879..15898)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(15880..15898)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(15908..15925)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(15908..15924)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15917..15939)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(16065..16520)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(16202..16223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     16414..16433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        16547..16651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(16652..16720,16721..17512)
                     /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     complement(16870..16889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gagccccagc tggttctttc
     1081 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
     1141 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     1201 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     1261 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     1321 ggctgatcag cgagctctag gatctgcatt ccaccactgc tcccattcat cagttccata
     1381 ggttggaatc taaaatacac aaacaattag aatcagtagt ttaacacatt atacacttaa
     1441 aaattttata tttaccttag agctttaaat ctctgtaggt agtttgtcca attatgtcac
     1501 accacagaag taaggttcct tcacaaagag atcgcctgac acgatttcct gcacaggctt
     1561 gagccatata ctcatacatc gcatcttggc cacgttttcc acgggtttca aaattaatct
     1621 caagttctac gcttaacgct ttcgcctgtt cccagttatt aatatattca acgctagaac
     1681 tcccctcagc gaagggaagg ctgagcacta cacgcgaagc accatcaccg aaccttttga
     1741 taaactcttc cgttccgact tgctccatca acggttcagt gagacttaaa cctaactctt
     1801 tcttaatagt ttcggcatta tccactttta gtgcgagaac cttcgtcagt cctggatacg
     1861 tcactttgac cacgcctcca gcttttccag agagcgggtt ttcattatct acagagtatc
     1921 ccgcagcgtc gtatttattg tcggtactat aaaacccttt ccaatcatcg tcataatttc
     1981 cttgtgtacc agattttggc ttttgtatac ctttttgaat ggaatctaca taaccaggtt
     2041 tagtcccgtg gtacgaagaa aagttttcca tcacaaaaga tttagaagaa tcaacaacat
     2101 catcaggatc catggcgagg acctgcaggt cgaaaggccc ggagatgagg aagaggagaa
     2161 cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg
     2221 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag
     2281 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac
     2341 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc
     2401 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg
     2461 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt
     2521 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc ccagcggggc
     2581 tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc ggtagaattt
     2641 cgacggtatc gataagctag atgtcctgaa atatttctat cagaacaagg tagtataaag
     2701 ctggtaggta tacaaaacgc tagactagtt tctatccctg acccttaatc tgctagtata
     2761 tccgtaggaa gttgcttaag tgccactagt accaacagcc tctgttagag tgaacagtag
     2821 attttaatga agggccaata aagaccactt aagactactg actgaattaa aaatccaccc
     2881 agtggagtta gtattggtta tagttatata acaaaatatc caacttagcc aggcgtggtg
     2941 gtgcacgcct ttaatcccgg cactcaggag gcagaggcag gcggatttct gagttcgagg
     3001 ccagcctggt ctacaaagtg agttccagga cagccagggc tatacagaga aaccctgtct
     3061 cgaaaaacca aaaaaaaaaa aaaaaaggaa aaaaaatatg gcttgaaaac atgctattgt
     3121 tgtcaaaagg agatccaaac aagtatttct ttgaacaaaa attatgtaga aaacagggtg
     3181 ataaagaaca agggcttttt aaaaaaggcc agcattattt gagatataca agtgactcca
     3241 attcaagcct aaaggccact caatgctcac taacagtgct aatcagaatc aattccttag
     3301 cccagtcaaa aatagtcctg aatatcttct ggataaacat gtcctccaac atataaacta
     3361 gtatacacta acaaaacgtc tcaacttcaa ggtgaaatgc ttgactccta gacttgtgac
     3421 ccagcaaagt gctctatagg tagggttact aggtcagaga gtcttgcctg caaaccacaa
     3481 ttatacaatt ccaccaaatg taagacatgt aaaaatgact tatttagctc aggttttgaa
     3541 aattaaaaat caaacccaca aagacctaac tttcacatta agtacaaatg tttaacatat
     3601 ataacatgtt ttgccttgat atatgaaatc atatgacatc acctgacagg aaacttaagt
     3661 ttatttgaac aacaatcagc ctaaggtagg cctagcacat gatcaggata gtgcagggaa
     3721 acccaaagaa gtgcttctga gtataaacca agagacctcg aaatagcagc tttgttctgt
     3781 atctcatggg ctttaactat cacttgaaaa caatttcaca aatcacaaaa ccaacatttg
     3841 tttagtttcc aactctgcct attcaccgga gaaatccatg tcccaatttg atgggggaaa
     3901 acttgaatga aatattattt ggaatatttt aatatccctg tgtatatgca gatggtttaa
     3961 agacaggtaa ccttaactgt agtttcttgg ttgtctgcct tgttttttga aatatgctct
     4021 caccaggagc ctgccaagta actactcttg tgtgctcagc aagtcctagg gatcccttaa
     4081 gcatgctcta acaggcctgg cttttttttt taaaaagtag gttgcggtga taataattca
     4141 agttttcatc acttgaaaca cattttacca actatcaccc aagctccctt agtgtgtccc
     4201 ctataaaaga atctgacctg caagttccaa aagtattact acattttaac cttagtattc
     4261 aggagacaaa agcaggcaga actttagtct acatatcttg ttgcagatca ggcagagata
     4321 catagtgaga ccctgcctca ggaaaagcta tcatttttat attcacagta agcaaattaa
     4381 cattaaaagt cagaaattct taatttgact ttggctgtga agaatttgga ttctctggat
     4441 ttgaaactag aaaatctgat taagcgatgc aacagtttct gtatactaaa actctaagac
     4501 ccttggttct aaagatacca catttaaaaa aaaagttaag tataagacaa aaagccttaa
     4561 tagctctttt ataggggagg gactcattta atattagtcc acctcactcc tcataactat
     4621 tttatgaggt ttattatata acccattttt taaaaaaaaa attaaaggac aattttagtg
     4681 tttgaaagat ttcccaaccc caccctggaa atcaggctgc aaatctcagc agctgccctt
     4741 agaaggcaaa aagccatgcc atttaagcca tgggaagtta gtagcaaaca agagaccaca
     4801 tggcaggaag gaggctttaa agaaagccca cagtgtcaaa agacccaacc aacagcagag
     4861 acagaaaaaa taatacccaa atctcatcag aaaggtagac ggatttagcc acatccatag
     4921 tggctcatta gggaatgctt caattataat tttatgaaat ctcagagccc ccctccccaa
     4981 cagggtttct gtgtgtattc ctggctatcc tagatagaac ttgatgtgta gaccaggctg
     5041 ggctcaaact cagagatctg cctgtctctg cctccagagt gctgggatta aaggcataaa
     5101 ggcaagcacc accactggct ggctaaactc tggccctaca tttttgaaat ccaaaattat
     5161 cattacttgc tttcatgttt agtaatggct gcagacttag ctttcagctt tgtatagatg
     5221 aagcacaaaa gttaagtaaa gctagcagtt taaacaaata ggatactaga agttcaattt
     5281 ttatttcttt attgcttagt gtaagagaaa agacattaaa gtctgcattg gtcttctgta
     5341 tcccacaagt ctgcagttat ggctcctctg tccacagtta cacttcaaat aaaaattagt
     5401 tccttggtaa catttccagt gcttaaagag taatttgatt attggctacc atattggaac
     5461 aaacacaaag tatttcatta cttcagtaaa ttctactgga gacagtattc tctttaaaaa
     5521 aaaaaaaaag aagaagaagg aggatttatt tatttaatgt gaatacactt gtggtcttca
     5581 gacacaccag aagagggcat cagatcccat tacagatggt tgtgagccac catgtggttg
     5641 ctgggaattg aactcaggac ttctggaaga gcagtctgtg ctcttgaact ccgagccacc
     5701 tctccagccc ctggatagta ttattataga gcacaagcac acacaacaca acaacctaaa
     5761 aaccaagctt cacaaataca tttgttgctt aatcatttta cttgtgtttg gctacaagtc
     5821 aagcaaaatt ataggtcctg aagaagcttg gcaaaatcac atttagacca gcaataacgt
     5881 gtagaatgcc atgagtcaag ccagtccaag agaaagcact gcttaaatag aaaaaataca
     5941 agctcacaag accttaggtc aggaaagaca acaaacacct gaactttgca ttccaaaagg
     6001 aaccaccttt tacagatgtg tacaatgaca aaattttagt aagcagtaat caataccatg
     6061 tggctcaata atgaaatata ccttttaatg tcttaaaata tcagcctggc aatatgtaag
     6121 atacatcagg taatattggg ggaggagaca tccacctgga aaccattaat ggttaatagc
     6181 tcagtttata aatggagaaa aaggagagag gcattcatgg gagtggaaag ttaagcttta
     6241 ggttggattc tcaatacatc tattgttgta aaactccctg gactgagaat aggcccaaat
     6301 gtggaacacc acctgacggg agaggtgata gacactgaaa attaaggatc aaggcaaagg
     6361 atcaacaaaa agtgtactaa ggagttataa aagaactgca gtgttgaggc cccagctaca
     6421 gcctcgattt gtggtgtatg taactaatct gtctggtttc atgagtcatc agacttctaa
     6481 gatcaggaaa gggaaaatgc caatgctctg tctaggggtt ggataagcca gtataataaa
     6541 tgaaaatggg gctaaaatga gtgttctaaa ataccttttg ataaggctgc agaaggagcg
     6601 ggagaaatgg atatgaagta ctgggctctt taaaaatgat taaaattctg cttacatagt
     6661 ctaactcgcg acactgtaat ttcatactgt agtaaggatc tcaagcagga gagtataaaa
     6721 ctcgggtgag catgtcttta atctacctcg atggaaaata ctccgaggcg gatcacaagc
     6781 aataataacc tgtagttttg ctgcataaaa ccccagatga ctacctatcc tcccattttc
     6841 cttatttgcc cctattaaaa aacttcccga caaaaccgaa aatctgtggg aagtcttgtc
     6901 cctccaattt tacacctgtt caattcccct gcaggacaac gcccacacac caggttagcc
     6961 tttaagcctg cccagaagac tcccgcccat cttctagccg gcgcgccgaa gttatattat
     7021 gtacctgact gatgaagttc ctatactttc tagagaatag gaacttcgaa gggttccgca
     7081 agctctagtc gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
     7141 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     7201 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     7261 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     7321 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctag agaattgatc
     7381 ccctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     7441 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     7501 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     7561 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     7621 cacgacgaga tcatcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg
     7681 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     7741 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     7801 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     7861 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     7921 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     7981 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     8041 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     8101 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     8161 tccatcttgt tcaatggccg atcccatggt ttagttcctc accttgtcgt attatactat
     8221 gccgatatac tatgccgatg attaattgtc aacaggctgc aggtcgaaag gcccggagat
     8281 gaggaagagg agaacagcgc ggcagacgtg cgcttttgaa gcgtgcagaa tgccgggcct
     8341 ccggaggacc ttcgggcgcc cgccccgccc ctgagcccgc ccctgagccc gcccccggac
     8401 ccaccccttc ccagcctctg agcccagaaa gcgaaggagc aaagctgcta ttggccgctg
     8461 ccccaaaggc ctacccgctt ccattgctca gcggtgctgt ccatctgcac gagactagtg
     8521 agacgtgcta cttccatttg tcacgtcctg cacgacgcga gctgcggggc gggggggaac
     8581 ttcctgacta ggggaggagt agaaggtggc gcgaaggggc caccaaagaa cggagccggt
     8641 tggcgcctac cggtggatgt ggaatgtgtg cgaggccaga ggccacttgt gtagcgccaa
     8701 gtgcccagcg gggctgctaa agcgcatgct ccagactgcc ttgggaaaag cgcctcccct
     8761 acccggtaga atttcgacga cctgcagcca agctagcttg gctggacgta aactcctctt
     8821 cagacctgaa gttcctatac tttctagaga ataggaactt cggaattcga tatcttaatt
     8881 aaagtagtgc gatcgcagta gtggccggcc ggttctttcc gcctcagaag ccatagagcc
     8941 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
     9001 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
     9061 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
     9121 gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc gagaagctta
     9181 agcgtagtca ggcacgtcat aagggtaccc cccggcatag tctggcacat cgtatgggta
     9241 accacccgcg taatcgggga cgtcataggg atagccggat cccttgtaca gctcgtccat
     9301 gccgtgagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     9361 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
     9421 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     9481 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggccat
     9541 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     9601 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     9661 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     9721 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     9781 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     9841 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     9901 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     9961 caccaccccg gtgaacagct cctcgccctt gctcatggtg gcccgcggta gcgctagccc
    10021 acctccgccg ctgcccccac ctccagatcc acccccgccg gagcctccac ccccagatcc
    10081 cttagcgctg gtgtacttgg tgacggcctt agtaccctcg gacacggcgt gcttggccaa
    10141 ctccccaggc agcagcaggc gcacggccgt ctggatctcc ctggaggtga tggtcgagcg
    10201 cttgttgtaa tgcgccaggc gggaagcctc acctgcgatg cgctcgaaaa tgtcgttcac
    10261 aaacgaattc atgatgccca tggccttgga cgaaatgccg gtgtcagggt ggacctgctt
    10321 cagaaccttg tacacataga tggaatagct ctccttgcgg ctgcgcttgc gcttcttgcc
    10381 gcctttcttc tgcgccttag tcaccgcctt cttggagccc tttttcgggg cgggagcaga
    10441 cttcgctggc tctggcatgg tgatattcag atctcctggg ccaggattct cctcgacgtc
    10501 accgcatgtt agcagacttc ctctgccctc tccactgcct cgcgtaccct ggaagtacag
    10561 gttctcgcta ctattaagat cctcctcgga tattaacttc tgctcaccgt tgaggtcttc
    10621 ctcactgatc aatttctgtt ctccattcaa gtcctcttca gaaatgagct tttgctcaga
    10681 ccccttgtac agctcgtcca tgccgtacag gaacaggtgg tggcggccct cggagcgctc
    10741 gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tggtgtccac
    10801 gtagtagtag ccgggcagtt gcacgggctt cttggccatg tagatggtct tgaactccac
    10861 caggtagtgg ccgccgtcct tcagcttcag ggcctggtgg atctcgccct tcagcacgcc
    10921 gtcgcggggg tacaggcgct cggtggaggc ctcccagccc atggtcttct tctgcattac
    10981 ggggccgtcg ggggggaagt tggtgccgcg catcttcacc ttgtagatca gcgtgccgtc
    11041 ctgcagggag gagtcctggg tcacggtcac cagaccgccg tcctcgaagt tcatcacgcg
    11101 ctcccacttg aagccctcgg ggaaggacag cttcttgtaa tcggggatgt cggcggggtg
    11161 cttcacgtac gccttggagc cgtacatgaa ctggggggac aggatgtccc aggcgaaggg
    11221 cagggggccg cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc
    11281 ctcgccctcg ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgcgcacctt
    11341 gaagcgcatg aactctttga tgacggccat gttgttgtcc tcggaggagg cggtgccgga
    11401 gctgccgctg ccggtgctgc cggtgccatg ccccaggaac aggtggtggc ggccctcgga
    11461 gcgctcgtac tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccagcttggt
    11521 gtccacgtag tagtagccgg gcagttgcac gggcttcttg gccatgtaga tggtcttgaa
    11581 ctccaccagg tagtggccgc cgtccttcag cttcagggcc tggtggatct cgcccttcag
    11641 cacgccgtcg cgggggtaca ggcgctcggt ggaggcctcc cagcccatgg tcttcttctg
    11701 cattacgggg ccgtcggggg ggaagttggt gccgcgcatc ttcaccttgt agatcagcgt
    11761 gccgtcctgc agggaggagt cctgggtcac ggtcaccaga ccgccgtcct cgaagttcat
    11821 cacgcgctcc cacttgaagc cctcggggaa ggacagcttc ttgtaatcgg ggatgtcggc
    11881 ggggtgcttc acgtacgcct tggagccgta catgaactgg ggggacagga tgtcccaggc
    11941 gaagggcagg gggccgccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg
    12001 gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg ttcatggagc cctccatgcg
    12061 caccttgaag cgcatgaact ctttgatgac ctcctcgccc ttgctcacca tggcgcgccg
    12121 cgacgaaggg ccgcagccca tggtggcggc cgcaataact tcgtataatg tatgctatac
    12181 gaagttatac tagcttggct gcaggtcgac agatccagac atgataagat acattgatga
    12241 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
    12301 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
    12361 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
    12421 cctctacaaa tgtggtatgg ctgattatga tccagacatg ataagataca ttgatgagtt
    12481 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
    12541 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
    12601 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
    12661 ctacaaatgt ggtatggctg attatgatcc agacatgata agatacattg atgagtttgg
    12721 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
    12781 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
    12841 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
    12901 caaatgtggt atggctgatt atgatcatgt ggatccggaa cccttaatat aacttcgtat
    12961 aatgtatgct atacgaagtt attaggtccc tcgaagaggt tcactagtgt cgagcccgcg
    13021 gtatcaaatc aattctttgc caaaatgatg agacagcaca ataaccagca cgttgcccag
    13081 gagctgtagg aaaaagaaga aggcatgaac atggttagca gaggctctag agccgccggt
    13141 cacacgccag aagccgaacc ccgccctgcc ccgtcccccc cgaaggcagc cgtccccccg
    13201 cggacagccc cgaggctgga gagggagaag gggacggcgg cgcggcgacg cacgaaggcc
    13261 ctccccgccc atttccttcc tgccggcgcc gcaccgcttc gccccgcgcc cgctagaggg
    13321 ggtgcggcgg cgcctcccag atttcggctc cgcacagatt tgggacaaag gaagtccctg
    13381 cgccctctcg cacgattacc ataaaaggca atggctgcgg ctcgccgcgc ctcgacagcc
    13441 gccggcgctc cgggggccgc cgcgcccctc ccccgagccc tccccggccc gaggcggccc
    13501 cgccccgccc ggcaccccca cctgccgcca ccccccgccc ggcacggcga gccccgcgcc
    13561 acgccccgta cggagccccg cacccgaagc cgggccgtgc tcagcaactc ggggaggggg
    13621 gtgcaggggg ggttgcagcc cgaccgacgc gcccacaccc cctgctcacc cccccacgca
    13681 cacaccccgc acgcagcctt tgttcccctc gcagcccccc ccgcaccgcg gggcaccgcc
    13741 cccggccgcg ctcccctcgc gcacactgcg gagcgcacaa agccccgcgc cgcgcccgca
    13801 gcgctcacag ccgccgggca gcgcggagcc gcacgcggcg ctccccacgc acacacacac
    13861 gcacgcaccc cccgagccgc tccccccgca caaagggccc tcccggagcc cctcaaggct
    13921 ttcacgcagc cacagaaaag aaacaagccg tcattaaacc aagcgctaat tacagcccgg
    13981 aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc agagccgggg
    14041 cgggcggcgc gaggcggcgg cggagcgggg cacggggcga aggcagcgcg cagcgactcc
    14101 cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc cgccgcctcg ccataaaagg
    14161 aaactttcgg agcgcgccgc tctgattggc tgccgccgca cctctccgcc tcgccccgcc
    14221 ccgcccctcg ccccgccccg ccccgcctgg cgcgcgcccc cccccccccc ccgcccccat
    14281 cgctgcacaa aataattaaa aaataaataa atacaaaatt gggggtgggg aggggggggg
    14341 agatggggag agtgaagcag aacgtggggc tcacctcgac catggtaata gcgatgacta
    14401 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
    14461 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
    14521 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
    14581 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
    14641 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
    14701 ggctatgaac taatgacccc gtaattgatt actattaata actagattcc tgcagaattg
    14761 tttaaacggt tataaggcta gaaagactgg agttgcagat cacgagggaa gagggggaag
    14821 ggattctccc aggcccaggg cggtcctcag aagccaggag gcagcagaga actcccagaa
    14881 aggtattgca acactcccct cccccctccg gagaagggtg cggccttctc cccgcctact
    14941 ccactgcagc tcccttactg ataacaactc agagcgactt tgggagagca agtgcttcct
    15001 gcctccaaaa cagcccaact gagccctcgc tccttccctc cactccccgg agtgcgcgat
    15061 ggaggtctgg ctcagcacgc ccctcttgag gcaactcaag tcggaaacgt gcttgcaccc
    15121 gccccgcagc cgctcagccc tactgcccgt ccccgccccc agcgcgcgct tcctgccacg
    15181 ttgcgcaggg gcgcgcggcc agactctgcg gcgcggggcc gaggggaggg ccggaacctg
    15241 ggagcgcctc ctcgccgccc ccgctggccg gcggatggac tcaacttgca cgaacacgag
    15301 ccaatggcaa gggccagttt tctgggcccc gagagccaat cagacgacga ggcccggccg
    15361 gcggcgggta aaacgactcc cccagaggaa ggggagggtg ggcggccgct cgcgcgcgag
    15421 ctactttcgc tgaccctccc tcccctcccc cgccccgcca gaggccgacc gcgcccgcac
    15481 gtccagctcg cctcacccca cctacctccc gccccaccca gtgggcagag cgaggctgcc
    15541 ggcggctgcg cactccggct gccgttaact gacaggcgcc ttacgccaac caaaacacgc
    15601 catttgtgtt ttcacacacg gcgggaggaa aagaagccaa tcagcgacga gacgtcggcc
    15661 ggaagcgctc ctccgctgcc cccccccccc ccgagccatg gccgcgtccg gtggagactt
    15721 ttccgctccc ttctccctcc ccctccggtt gctgcagggc ggaccgcatt cctgcccacc
    15781 acccgcttgc cccttccagc gtcacgactc gtacccggct gtctcacaga acggctccac
    15841 cacgctcgga gggcctgccg cggtggagct ccaattcgcc ctatagtgag tcgtattacg
    15901 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
    15961 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
    16021 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaaattg taagcgttaa
    16081 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
    16141 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
    16201 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
    16261 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
    16321 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
    16381 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
    16441 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
    16501 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
    16561 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
    16621 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
    16681 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
    16741 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
    16801 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
    16861 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
    16921 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
    16981 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
    17041 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
    17101 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
    17161 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
    17221 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
    17281 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
    17341 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
    17401 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
    17461 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
    17521 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
    17581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
    17641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
