lentiMPH v2

LOCUS       lentiMPH v2               11739 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  lenti vector encoding the MS2-P65-HSF1 activator helper complex with
            a 2A Hygro resistance marker (EF1a-MS2-p65-HSF1-2A-Hygro-WPRE). This
            version has a higher virus titer..
ACCESSION   .
VERSION     .
KEYWORDS    lentiMPH v2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11739)
  AUTHORS   Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham
            MD, Sanjana NE, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout and transcriptional activation 
            screening.
  JOURNAL   Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. 
            Epub 2017 Mar 23.
  PUBMED    28333914
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|lentiMPH v2 |
FEATURES             Location/Qualifiers
     source          1..11739
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    488..613
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             660..840
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(858..1056)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(883..903)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1058..1437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1609..1628
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1703..1807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1808..2668
                     /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(2026..2045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2839..3427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3328..3347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3581..3598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3715..3736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3751..3781
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3789..3805
                     /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     3794..3816
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3813..3829
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3813..3829
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3878..3999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3889..3908)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3943..3962
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4129..4503)
                     /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"
     promoter        complement(4522..4569)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4617..4946)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4631..4766
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4685..4704)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4931..4951
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4960..5388)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5070..5091)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5282..5301
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5434..5658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5647..5664
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             5690..5870
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     6375..6391
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6376..6392
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6475..7653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6706..7644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7601..7621
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7660..7669
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7669..8055
                     /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"
     CDS             8110..8130
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8146..8688
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             8713..9084
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             9100..9153
                     /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             9157..10176
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     misc_feature    10216..10804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10269..10289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10687..10698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    10857..10974
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    11501..11734
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
       61 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
      121 tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta cttttgtttt
      181 gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca
      241 cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa
      301 gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag
      361 gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt
      421 ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct
      481 taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg
      541 gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc
      601 tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact
      661 gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca
      721 caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag
      781 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
      841 agtacaggca aaacgcgctg cttatataga cctcccaccg tacacgccta ccgcccattt
      901 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
      961 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     1021 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     1081 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     1141 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     1201 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     1261 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     1321 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     1381 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcaac
     1441 gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta agcagattct
     1501 tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact
     1561 cagcgacctc caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc
     1621 agattgtact gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg
     1681 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1741 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1801 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1861 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1921 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1981 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2041 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2101 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2161 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2221 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2281 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2341 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2401 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2461 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2521 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2581 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2641 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2701 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2761 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3541 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3601 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3661 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3721 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3781 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     3841 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
     3901 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3961 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
     4021 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     4081 caacctttca tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc
     4141 ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca
     4201 cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac
     4261 gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt
     4321 gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac
     4381 cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa
     4441 ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc
     4501 catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta
     4561 attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg
     4621 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag
     4681 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     4741 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     4801 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4861 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4921 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
     4981 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5041 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5101 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5161 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5221 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5281 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5341 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc
     5401 tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat
     5461 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     5521 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     5581 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     5641 ggcaactaga aggcacagtc gaggctgatc agcgggttta aacgggccct gctagagatt
     5701 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     5761 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     5821 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacagtcc
     5881 ggatgcagct ctcgggccat gtgatgaaat gctaggcggc tgtcaaacct ccactctaat
     5941 acttctctct ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gcgggtgttc
     6001 tctccttcat tggcttcttc taccttctct tgctcaactg gtactagctt gtagcaccat
     6061 ccaaaggtca gtggatatct gatccctggc cctggtgtgt agttctgcca atcagggaag
     6121 tagccttgtg tgtggtagat ccacagatca aggatatctt gtcttcgttg ggagtgaatt
     6181 agcccttcca gtcccccctt ttcttttaaa aagtggctaa gatctacagc tgccttgtaa
     6241 gtcattggtc ttaaaggtac ctgaggtgtg actggaaaac ccacctcctc ctcctcttgt
     6301 gcttctagcc aggcacaatc agcattggta gctgctgtat tgctacttgt gattgctcca
     6361 tgtttttcta ggtctcgagg tcgacggtat cgattgcaaa gatggataaa gttttaaaca
     6421 gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc
     6481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     6541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     6601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     6661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
     6721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
     6781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
     6841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
     6901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     6961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     7021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     7081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     7141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     7201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     7261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg aaggacgcgg
     7321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     7381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     7441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     7501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     7561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     7621 caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatggc ttcaaacttt
     7681 actcagttcg tgctcgtgga caatggtggg acaggggatg tgacagtggc tccttctaat
     7741 ttcgctaatg gggtggcaga gtggatcagc tccaactcac ggagccaggc ctacaaggtg
     7801 acatgcagcg tcaggcagtc tagtgcccag aagagaaagt ataccatcaa ggtggaggtc
     7861 cccaaagtgg ctacccagac agtgggcgga gtcgaactgc ctgtcgccgc ttggaggtcc
     7921 tacctgaaca tggagctcac tatcccaatt ttcgctacca attctgactg tgaactcatc
     7981 gtgaaggcaa tgcaggggct cctcaaagac ggtaatccta tcccttccgc catcgccgct
     8041 aactcaggta tctacagcgc tggaggaggt ggaagcggag gaggaggaag cggaggagga
     8101 ggtagcggac ctaagaaaaa gaggaaggtg gcggccgctg gatccccttc agggcagatc
     8161 agcaaccagg ccctggctct ggcccctagc tccgctccag tgctggccca gactatggtg
     8221 ccctctagtg ctatggtgcc tctggcccag ccacctgctc cagcccctgt gctgacccca
     8281 ggaccacccc agtcactgag cgctccagtg cccaagtcta cacaggccgg cgaggggact
     8341 ctgagtgaag ctctgctgca cctgcagttc gacgctgatg aggacctggg agctctgctg
     8401 gggaacagca ccgatcccgg agtgttcaca gatctggcct ccgtggacaa ctctgagttt
     8461 cagcagctgc tgaatcaggg cgtgtccatg tctcatagta cagccgaacc aatgctgatg
     8521 gagtaccccg aagccattac ccggctggtg accggcagcc agcggccccc cgaccccgct
     8581 ccaactcccc tgggaaccag cggcctgcct aatgggctgt ccggagatga agacttctca
     8641 agcatcgctg atatggactt tagtgccctg ctgtcacaga tttcctctag tgggcaggga
     8701 ggaggtggaa gcggcttcag cgtggacacc agtgccctgc tggacctgtt cagcccctcg
     8761 gtgaccgtgc ccgacatgag cctgcctgac cttgacagca gcctggccag tatccaagag
     8821 ctcctgtctc cccaggagcc ccccaggcct cccgaggcag agaacagcag cccggattca
     8881 gggaagcagc tggtgcacta cacagcgcag ccgctgttcc tgctggaccc cggctccgtg
     8941 gacaccggga gcaacgacct gccggtgctg tttgagctgg gagagggctc ctacttctcc
     9001 gaaggggacg gcttcgccga ggaccccacc atctccctgc tgacaggctc ggagcctccc
     9061 aaagccaagg accccactgt ctcctgtaca ggcagtggag agggcagagg aagtctgcta
     9121 acatgcggtg acgtcgagga gaatcctggc ccaaccatga aaaagcctga actcaccgct
     9181 acctctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     9241 tccgagggcg aagaatctcg ggctttcagc ttcgatgtgg gagggcgtgg atatgtcctg
     9301 cgggtgaata gctgcgccga tggtttctac aaagatcgct atgtttatcg gcactttgca
     9361 tccgccgctc tccctattcc cgaagtgctt gacattgggg agttcagcga gagcctgacc
     9421 tattgcatct cccgccgtgc acagggtgtc accttgcaag acctgcctga aaccgaactg
     9481 cccgctgttc tccagcccgt cgccgaggcc atggatgcca tcgctgccgc cgatcttagc
     9541 cagaccagcg ggttcggccc attcggacct caaggaatcg gtcaatacac tacatggcgc
     9601 gatttcatct gcgctattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     9661 accgtcagtg cctccgtcgc ccaggctctc gatgagctga tgctttgggc cgaggactgc
     9721 cccgaagtcc ggcacctcgt gcacgccgat ttcggctcca acaatgtcct gaccgacaat
     9781 ggccgcataa cagccgtcat tgactggagc gaggccatgt tcggggattc ccaatacgag
     9841 gtcgccaaca tcttcttctg gaggccctgg ttggcttgta tggagcagca gacccgctac
     9901 ttcgagcgga ggcatcccga gcttgcagga tctcctcggc tccgggctta tatgctccgc
     9961 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
    10021 gctcagggtc gctgcgacgc aatcgtccgg tccggagccg ggactgtcgg gcgtacacaa
    10081 atcgcccgca gaagcgctgc cgtctggacc gatggctgtg tggaagtgct cgccgatagt
    10141 ggaaacagac gccccagcac tcgtcctagg gcaaaggatc tgcagtaatg agaattcgat
    10201 atcaagctta tcggtaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
    10261 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
    10321 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
    10381 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
    10441 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
    10501 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
    10561 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
    10621 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
    10681 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
    10741 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
    10801 ccgcatcgct agctaattaa ccaaactgga tctctgctgt ccctgtaata aacccgaaaa
    10861 ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt tgctattatg
    10921 tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt aaaattgtgg
    10981 atgaatactg ccatttgtct gcagaattgg cgcacgcagt gccgatccgt tcactaatcg
    11041 aatggatctg tctctgtctc tctctccacc ttcttcttct attccttcgg gcctgtcggg
    11101 tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct
    11161 attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat
    11221 tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc
    11281 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
    11341 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
    11401 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
    11461 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
    11521 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
    11581 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
    11641 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
    11701 tgctgctccc aagaacccaa ggaacaaagc tcctattcc
//