LOCUS       sequence-384402-       12559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus, expresses Cre recombinase and mScarlet-SIINFEKL, with 
            sgRNA cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12559)
  AUTHORS   Freed-Pastor WA, Lambert LJ, Ely ZA, Pattada NB, Bhutkar A, Eng G, 
            Mercer KL, Garcia AP, Lin L, Rideout WM 3rd, Hwang WL, Schenkel JM, 
            Jaeger AM, Bronson RT, Westcott PMK, Hether TD, Divakar P, Reeves 
            JW, Deshpande V, Delorey T, Phillips D, Yilmaz OH, Regev A, Jacks T
  TITLE     The CD155/TIGIT axis promotes and maintains immune evasion in 
            neoantigen-expressing pancreatic cancer.
  JOURNAL   Cancer Cell. 2021 Oct 11;39(10):1342-1360.e14. doi: 
            10.1016/j.ccell.2021.07.007. Epub 2021 Aug 5.
  PUBMED    34358448
REFERENCE   2  (bases 1 to 12559)
  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"; doi: 
            "10.1016/j.ccell.2021.07.007"; journalName: "Cancer Cell"; date: 
            "2021-10-11- 11"; volume: "39"; issue: "10"; pages: "1342-1360.e14"
COMMENT     Sequence Label: LV-U6-sgRNAfiller-PGK-Cre-EFS-mScarletSIIN
FEATURES             Location/Qualifiers
     source          1..12559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2638..2886
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2638..2658
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2809..2828
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4768..4843
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4850..5349
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4885..4908)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5282..5301
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5389..5409
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5410..6438
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     promoter        6494..6705
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6750..6759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6756..7451
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(6889..6907)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7157..7176
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7539..7562
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
     misc_feature    7651..8239
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7704..7724)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8122..8133)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8311..8544
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8570..8587)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8576..8800
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8846..9274
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8933..8952)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9143..9164
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9283..9303)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9288..9617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9468..9603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9530..9549
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9665..9712
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9731..10105
                     /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    10235..10356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10272..10291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10326..10345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10405..10421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10405..10421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10418..10440)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10429..10445
                     /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(10453..10459,10460..10477,10478..10483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10498..10519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10636..10653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10807..11395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10887..10906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11566..12357,12358..12426))
                     /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     12189..12208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12427..12531)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgat cagtgtgagg gagtgtaaag ctggtttgag
     2641 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     2701 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     2761 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     2821 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     2881 aacaccggag acggttgtaa atgagcacac aaaatacaca tgctaaaata ttatattcta
     2941 tgacctttat aaaatcaacc aaaatcttct ttttaataac tttagtatca ataattagaa
     3001 tttttatgtt cctttttgca aacttttaat aaaaatgagc aaaataaaaa aacgctagtt
     3061 ttagtaactc gcgttgtttt cttcaccttt aataatagct actccaccac ttgttcctaa
     3121 gcggtcagct cctgcttcaa tcattttttg agcatcttca aatgttctaa ctccaccagc
     3181 tgctttaact aaagcattgt ctttaacaac tgacttcatt agtttaacat cttcaaatgt
     3241 tgcacctgat tttgaaaatc ctgttgatgt tttaacaaat tctaatccag cttcaacagc
     3301 tatttcacaa gctttcatga tttcttcttt tgttaataaa caattttcca taatacattt
     3361 aacaacatgt gatccagctg ctttttttac agctttcatg tcttctaaaa ctaattcata
     3421 atttttgtct tttaatgcac caatatttaa taccatatca atttctgttg caccatcttt
     3481 aattgcttca gaaacttcga atgcttttgt agctgttgtg catgcaccta gaggaaaacc
     3541 tacaacattt gttattccta catttgtgcc ttttaataat tctttacaat agcttgttca
     3601 atatgaatta acacaaactg ttgcaaaatc aaattcaatt gcttcatcac ataattgttt
     3661 aatttcagct ttcgtagcat cttgttttaa taatgtgtga tctatatatt tgtttagttt
     3721 cattttttct cctatatatt catttttaat tttaattctt taataatttc gtctacttta
     3781 actttagcgt tttgaacaga ttcaccaaca cctataaaat aaatttttag tttaggttca
     3841 gttccacttg ggcgaacagc aaatcatgac ttatcttcta aataaaattt tagtaagtct
     3901 tgtcctggca tattatacat tccatcgatg tagtcttcaa cattaacaac tttaagtcca
     3961 gcaatttgag ttaagggtgt tgctctcaat gatttcatta atggttcaat ttttaatttc
     4021 ttttcttctg gtttaaaatt caagtttaaa gtgaaagtgt aatatgcacc catttcttta
     4081 aataaatctt ctaaatagtc tactaatgtt ttattttgtt ttttataaaa tcaagcagcc
     4141 tctgctatta atatagaagc ttgtattcca tctttatctc tagctgagtc atcaattaca
     4201 tatccataac tttcttcata agcaaaaaca aaatttaatc cgttatcttc ttctttagca
     4261 atttctctac ccattcattt aaatccagtt aaagttttta caatattaac tccatatttt
     4321 tcatgagcga ttctatcacc caaatcactt gttacaaaac ttgaatatag agccggattt
     4381 tttggaatgc tatttaagcg ttttagattt gataattttc aatcaattaa aattggtcct
     4441 gtttgatttc catctaatct tacaaaatga ccatcatgtt ttattgccat tccaaatctg
     4501 tcagcatctg ggtcattcat aataataata tctgcatcat gtttaatacc atattcaagc
     4561 ggtatttttc atgcaggatc aaattctgga tttggattta caacattttt aaatgtttca
     4621 tcttcaaatg catgctcttc aacctcaata acgttatatc ctgattcacg taatattttt
     4681 ggggtaaatt tagttcctgt tccattaact gcgctaaaaa taatttttaa atctttttta
     4741 gcttcttgct cttttttgta cgtctctgtt ttagagctag aaatagcaag ttaaaataag
     4801 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ggtaggggag
     4861 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     4921 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     4981 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     5041 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tgtcactagt
     5101 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     5161 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     5221 ggcgggctca ggggcgggct caggggcggt gcgggcgccc gaaggtcctc cggaggcccg
     5281 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     5341 ggcctttcgc taactcgaac gctagctgtg cgatcgtttg ccaccatgcc caagaagaag
     5401 aggaaggtgt ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca
     5461 acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct
     5521 gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg
     5581 aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt
     5641 caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt
     5701 catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg
     5761 cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa
     5821 cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata
     5881 cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc
     5941 aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc
     6001 agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact
     6061 aaactggtcg agcgatggat ttccgtatct ggtgtagctg atgatccgaa taactacctg
     6121 ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact
     6181 cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac
     6241 tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat
     6301 atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta
     6361 aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg
     6421 ctggaagatg gcgattagaa accgctgttc ctaggaatcc cgaggccttc gagtggctcc
     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 cacaggtgtc gtgacgcggg
     6721 acccgacatt agcgctacag cttaagcggg ccaccatggt gagcaagggc gaggcagtga
     6781 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
     6841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     6901 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
     6961 gctccagggc cttcaccaag caccccgccg acatccccga ctactataag cagtccttcc
     7021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
     7081 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
     7141 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
     7201 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
     7261 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
     7321 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
     7381 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
     7441 agctgtacaa ggagagcctg aagatatctc aagctgtcca tgcagcacat gcagaaatca
     7501 atgaagcagg cagagaggtg gtaggggagc agctcgagag tataatcaac tttgaaaaac
     7561 tgtaaaaacg ttgttgtttg gggttgaatt actctacatg tttaagggtt ccggttccac
     7621 taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     7681 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     7741 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     7801 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     7861 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     7921 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     7981 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     8041 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     8101 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     8161 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     8221 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
     8281 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     8341 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     8401 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     8461 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     8521 agtcagtgtg gaaaatctct agcagggccc ggcgaaaccc gctgatcagc ctcgactgtg
     8581 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     8641 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     8701 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     8761 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     8821 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8881 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8941 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     9001 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     9061 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     9121 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     9181 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9241 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     9301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     9361 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     9421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     9481 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     9541 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     9601 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     9661 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     9721 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     9781 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     9841 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     9901 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     9961 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
    10021 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
    10081 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
    10141 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
    10201 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
    10261 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    10321 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
    10381 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
    10441 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
    10501 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
    10561 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
    10621 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
    10681 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    10741 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    10801 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    10861 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    10921 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    10981 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    11041 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    11101 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    11161 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    11221 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    11281 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    11341 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    11401 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    11461 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    11521 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    11581 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    11641 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    11701 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    11761 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    11821 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    11881 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    11941 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    12001 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    12061 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    12121 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    12181 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    12241 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    12301 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    12361 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    12421 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    12481 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    12541 ccgaaaagtg ccacctgac
//
