lentiMPH v2 (Neo)

LOCUS        lentiMPH v2 (Neo)       11505 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Modified version of lentiMPH v2, a lenti vector encoding the 
            MS2-P65-HSF1 activator helper complex and neo resistance marker 
            (EF1a-MS2-p65-HSF1-2A-Neo-WPRE)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11505)
  AUTHORS   Barger CJ, Chee L, Albahrani M, Munoz-Trujillo C, Boghean L, Branick
            C, Odunsi K, Drapkin R, Zou L, Karpf AR
  TITLE     Co-regulation and function of FOXM1/RHNO1 bidirectional genes in 
            cancer.
  JOURNAL   Elife. 2021 Apr 23;10. pii: 55070. doi: 10.7554/eLife.55070.
  PUBMED    33890574
REFERENCE   2  (bases 1 to 11505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-04-23"; volume: "10. pii: 55070. doi"; pages: " 
            10.7554/eLife.55070"
COMMENT     Sequence Label: lentiMPH v2 (Neo)
FEATURES             Location/Qualifiers
     source          1..11505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             43..223
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(249..266)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    255..479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      525..953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(612..631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     822..843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(962..982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        967..1296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1147..1282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1209..1228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1344..1391
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1410..1784
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1914..2035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1951..1970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2005..2024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2084..2100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2084..2100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2097..2119)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2108..2124
                     /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(2132..2138,2139..2156,2157..2162))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2177..2198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2315..2332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2486..3074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2566..2585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3245..4036,4037..4105))
                     /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     3868..3887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4106..4210)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4285..4304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4476..4855
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4857..5055
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5010..5030
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5073..5253
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5300..5425
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5918..6151
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6336..6380
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6678..6795
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6848..7436
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6954..6965
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7363..7383
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7461..8264)
                     /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="MSRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7572..7591)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8182..8201
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(8265..8318)
                     /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(8334..8705)
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             complement(8730..9272)
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             complement(9288..9308)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9363..9749)
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     regulatory      9749..9758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9765..10943)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9774..10712
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(9797..9817)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(11026..11042)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11027..11043)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
       61 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
      121 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
      181 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg
      241 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
      301 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
      361 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
      421 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
      481 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
      541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
      601 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
      661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
      721 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
      781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
      841 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
      901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
      961 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1021 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     1081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1201 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     1261 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     1321 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     1381 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     1441 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     1501 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     1561 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     1621 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     1681 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     1741 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     1801 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     1861 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     1921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     2041 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     2101 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2161 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2221 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2281 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2341 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2401 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2461 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2521 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2581 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2641 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2701 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2761 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2821 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2881 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2941 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3001 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3061 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3241 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3301 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3361 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3421 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3481 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3541 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3601 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3661 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3721 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3781 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3841 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3901 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3961 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4021 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4081 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4141 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4201 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     4261 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4321 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     4381 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     4441 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     4501 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     4561 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     4621 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     4681 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     4741 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     4801 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     4861 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     4921 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     4981 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     5041 ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag
     5101 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     5161 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     5221 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     5281 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     5341 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     5401 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     5461 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     5521 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     5581 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     5641 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     5701 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     5761 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     5821 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     5881 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     5941 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     6001 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     6061 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     6121 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     6181 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     6241 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     6301 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     6361 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     6421 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     6481 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     6541 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     6601 acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg
     6661 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     6721 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     6781 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gttaattagc
     6841 tagcgatgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga
     6901 cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg
     6961 gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc
     7021 agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc
     7081 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg
     7141 agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg
     7201 ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac
     7261 tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     7321 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     7381 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattaccg
     7441 ataagcttga tatcgaattc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     7501 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     7561 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     7621 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     7681 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
     7741 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     7801 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     7861 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     7921 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     7981 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     8041 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
     8101 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     8161 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     8221 ggagaacctg cgtgcaatcc atcttgttca atcattctag acattggacc agggttttct
     8281 tcaacatcac cacaagtgag gagagaacct ctaccttctc cgcttcctgt acaggagaca
     8341 gtggggtcct tggctttggg aggctccgag cctgtcagca gggagatggt ggggtcctcg
     8401 gcgaagccgt ccccttcgga gaagtaggag ccctctccca gctcaaacag caccggcagg
     8461 tcgttgctcc cggtgtccac ggagccgggg tccagcagga acagcggctg cgctgtgtag
     8521 tgcaccagct gcttccctga atccgggctg ctgttctctg cctcgggagg cctggggggc
     8581 tcctggggag acaggagctc ttggatactg gccaggctgc tgtcaaggtc aggcaggctc
     8641 atgtcgggca cggtcaccga ggggctgaac aggtccagca gggcactggt gtccacgctg
     8701 aagccgcttc cacctcctcc ctgcccacta gaggaaatct gtgacagcag ggcactaaag
     8761 tccatatcag cgatgcttga gaagtcttca tctccggaca gcccattagg caggccgctg
     8821 gttcccaggg gagttggagc ggggtcgggg ggccgctggc tgccggtcac cagccgggta
     8881 atggcttcgg ggtactccat cagcattggt tcggctgtac tatgagacat ggacacgccc
     8941 tgattcagca gctgctgaaa ctcagagttg tccacggagg ccagatctgt gaacactccg
     9001 ggatcggtgc tgttccccag cagagctccc aggtcctcat cagcgtcgaa ctgcaggtgc
     9061 agcagagctt cactcagagt cccctcgccg gcctgtgtag acttgggcac tggagcgctc
     9121 agtgactggg gtggtcctgg ggtcagcaca ggggctggag caggtggctg ggccagaggc
     9181 accatagcac tagagggcac catagtctgg gccagcactg gagcggagct aggggccaga
     9241 gccagggcct ggttgctgat ctgccctgaa ggggatccag cggccgccac cttcctcttt
     9301 ttcttaggtc cgctacctcc tcctccgctt cctcctcctc cgcttccacc tcctccagcg
     9361 ctgtagatac ctgagttagc ggcgatggcg gaagggatag gattaccgtc tttgaggagc
     9421 ccctgcattg ccttcacgat gagttcacag tcagaattgg tagcgaaaat tgggatagtg
     9481 agctccatgt tcaggtagga cctccaagcg gcgacaggca gttcgactcc gcccactgtc
     9541 tgggtagcca ctttggggac ctccaccttg atggtatact ttctcttctg ggcactagac
     9601 tgcctgacgc tgcatgtcac cttgtaggcc tggctccgtg agttggagct gatccactct
     9661 gccaccccat tagcgaaatt agaaggagcc actgtcacat cccctgtccc accattgtcc
     9721 acgagcacga actgagtaaa gtttgaagcc atggtggccg tacgtcacga cacctgaaat
     9781 ggaagaaaaa aactttgaac cactgtctga ggcttgagaa tgaaccaaga tccaaactca
     9841 aaaagggcaa attccaagga gaattacatc aagtgccaag ctggcctaac ttcagtctcc
     9901 acccactcag tgtggggaaa ctccatcgca taaaacccct ccccccaacc taaagacgac
     9961 gtactccaaa agctcgagaa ctaatcgagg tgcctggacg gcgcccggta ctccgtggag
    10021 tcacatgaag cgacggctga ggacggaaag gcccttttcc tttgtgtggg tgactcaccc
    10081 gcccgctctc ccgagcgccg cgtccttcat tttgagctcc ctgcagcagg gccgggaagc
    10141 ggccatcttt ccgctcacgc aactggtgcc gaccgggcca gccttgccgc ccagggcggg
    10201 gcgatacacg gcggcgcgag gccaggcacc agagcaggcc ggccagcttg agactacccc
    10261 cgtccgattc tcggtggccg cgctcgcagg ccccgcctcg ccgaacatgt gcgctgggac
    10321 gcacgggccc cgtcgccgcc cgcggcccca aaaaccgaaa taccagtgtg cagatcttgg
    10381 cccgcattta caagactatc ttgccagaaa aaaagcgtcg cagcaggtca tcaaaaattt
    10441 taaatggcta gagacttatc gaaagcagcg agacaggcgc gaaggtgcca ccagattcgc
    10501 acgcggcggc cccagcgccc aggccaggcc tcaactcaag cacgaggcga aggggctcct
    10561 taagcgcaag gcctcgaact ctcccaccca cttccaaccc gaagctcggg atcaagaatc
    10621 acgtactgca gccaggtgga agtaattcaa ggcacgcaag ggccataacc cgtaaagagg
    10681 ccaggcccgc gggaaccaca cacggcactt acctgtgttc tggcggcaaa cccgttgcga
    10741 aaaagaacgt tcacggcgac tactgcactt atatacggtt ctcccccacc ctcgggaaaa
    10801 aggcggagcc agtacacgac atcactttcc cagtttaccc cgcgccacct tctctaggca
    10861 ccggttcaat tgccgacccc tccccccaac ttctcgggga ctgtgggcga tgtgcgctct
    10921 gcccactgac gggcaccgga gccaattccc actcctttca agacctagaa ggtccattag
    10981 ctgcaaagat tcctctctgt ttaaaacttt atccatcttt gcaatcgata ccgtcgacct
    11041 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
    11101 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
    11161 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
    11221 gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
    11281 taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga
    11341 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa
    11401 gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat
    11461 gacccggaga gagaagtatt agagtggagg tttgacagcc gccta
//