LOCUS       Exported               11986 bp DNA     circular SYN 27-DEC-2021
DEFINITION  Express sgRNA and dCas9-KRAB from 3rd generation lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    Inducible tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11986)
  AUTHORS   Thakore PI, D'Ippolito AM, Song L, Safi A, Shivakumar NK, Kabadi AM,
            Reddy TE, Crawford GE, Gersbach CA
  TITLE     Highly specific epigenome editing by CRISPR-Cas9 repressors for 
            silencing of distal regulatory elements.
  JOURNAL   Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 
            2015 Oct 26.
  PUBMED    26501517
REFERENCE   2  (bases 1 to 11986)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 8, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.3630"; journalName: "Nat Methods"; date: "2015-12"; 
            volume: "12"; issue: "12"; pages: "1143-9"
FEATURES             Location/Qualifiers
     source          1..11986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     misc_feature    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     protein_bind    2364..2397
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     primer_bind     complement(2383..2407)
                     /label=CRISPRi  1st plasmid R
                     /note="color: black; sequence: tttttttttaataacttcgtataat; 
                     added: 2020-05-02"
     misc_RNA        complement(2408..2447)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: LEFT"
     primer_bind     2408..2447
                     /label=rTetR overhang F
                     /note="color: black; sequence: 
                     gcaccgactcggtgccactttttcaagttgataacggact; added: 
                     2020-11-19"
     primer_bind     2448..2469
                     /label=insert Fa
                     /note="color: black; sequence: ggagtaacccacccaagatctg; 
                     added: 2020-11-19"
     promoter        2469..3680
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     3517..3536
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
                     /note="color: black; sequence: TGAAGCTCCGGTTTTGAACT"
     CDS             3739..4485
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /note="This feature has 8 segments:
                        1: 3739 .. 4356 / #993366 / rTetR
                        2: 4357 .. 4362 / #cc99b2
                        3: 4363 .. 4398 / #993366 / VP16 minimal transcriptio...
                        4: 4399 .. 4401 / #cc99b2
                        5: 4402 .. 4437 / #993366 / VP16 minimal transcriptio...
                        6: 4438 .. 4440 / #cc99b2
                        7: 4441 .. 4476 / #993366 / VP16 minimal transcriptio...
                        8: 4477 .. 4485 / #cc99b2"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(4463..4485)
                     /label=insert Ra
                     /note="color: black; sequence: ttacccggggagcatgtcaaggt; 
                     added: 2020-11-19"
     primer_bind     complement(4486..4525)
                     /label=rTetR overhang R
                     /note="color: black; sequence: 
                     gggcctcaagatctggagccgacacgggttaattaattaa; added: 
                     2020-11-19"
     primer_bind     4486..4517
                     /label=TRE3GV overhang F
                     /note="color: black; sequence: 
                     ggccctcttaattaattaacccgtgtcggctccagatct; added: 2020-11-19"
     primer_bind     4518..4539
                     /label=insert F
                     /note="color: black; sequence: tgaggccctttcgtcttcactc; 
                     added: 2020-11-19"
     promoter        4540..4889
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    4548..4566
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4584..4602
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4620..4638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4656..4674
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4692..4710
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4728..4746
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4764..4782
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     primer_bind     4794..4803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     primer_bind     4828..4852
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
                     /note="color: black; sequence: AGCTCGTTTAGTGAACCGTCAGATC"
     primer_bind     complement(4890..4911)
                     /label=insert R
                     /note="color: black; sequence: gcttgatatcctgcagacgcgt; 
                     added: 2020-11-19"
     primer_bind     4910..4957
                     /label=tdTomato Full - 2nd attempt
                     /note="color: black; sequence: 
                     gcgaagcttgggctgcaggtcgactctagaGCCACCATGGTGAGCAAG; added: 
                     2020-11-19"
     primer_bind     4910..4939
                     /label=tdTomato overhang F 67.1
                     /note="color: black; sequence: 
                     gcgaagcttgggctgcaggtcgactctaga; added: 2020-11-19"
     primer_bind     complement(4912..4946)
                     /label=TRE3GV overhang R
                     /note="color: black; sequence: 
                     tggtggctctagagtcgacctgcagcccaagcttc; added: 2020-11-19"
     primer_bind     4940..4957
                     /label=insert F`b 52.6
                     /note="color: black; sequence: GCCACCATGGTGAGCAAG; added: 
                     2020-11-19"
     regulatory      4940..4945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             4946..6376
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    4967..4988
                     /label=Begining of tandom sequence
                     /note="color: #a6acb3"
     primer_bind     5296..5316
                     /label=qPCR F
                     /note="color: black; sequence: CACGCTGATCTACAAGGTGAA; 
                     added: 2020-12-13"
     primer_bind     complement(5352..5373)
                     /label=qPCR R
                     /note="color: black; sequence: CCCATGGTCTTCTTCTGCATTA; 
                     added: 2020-12-13"
     primer_bind     6022..6042
                     /label=qPCR F
                     /note="color: black; sequence: CACGCTGATCTACAAGGTGAA; 
                     added: 2020-12-13"
     primer_bind     complement(6078..6099)
                     /label=qPCR R
                     /note="color: black; sequence: CCCATGGTCTTCTTCTGCATTA; 
                     added: 2020-12-13"
     primer_bind     complement(6356..6376)
                     /label=insert R`b 52.4
                     /note="color: black; sequence: CTACTTGTACAGCTCGTCCAT; 
                     added: 2020-11-19"
     primer_bind     6356..6376
                     /label=tdTomato chk.2 F 52.4
                     /note="color: black; sequence: ATGGACGAGCTGTACAAGTAG; 
                     added: 2020-11-19"
     primer_bind     complement(6377..6424)
                     /label=tdTomato overhang 62
                     /note="color: black; sequence: 
                     tcgaattcataacttcgtataatgtatgctatacgaagttatctatta; added: 
                     2020-11-19"
     protein_bind    6383..6416
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    6441..7029
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(6494..6514)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     primer_bind     complement(6548..6567)
                     /label=GFP_SKL_chk_R- I'm using it to check tdTomato 49.7
                     /note="color: black; sequence: aagccatacgggaagcaata; added:
                     2020-11-19"
     CDS             complement(6912..6923)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 6911"
                     /translation="IEGR"
     primer_bind     complement(7032..7048)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(7033..7049)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
                     /note="color: black; sequence: TCGAGGTCGACGGTATC"
     LTR             7554..7734
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     primer_bind     complement(7760..7777)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    7766..7990
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      8036..8464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(8123..8142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     primer_bind     8333..8354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     primer_bind     complement(8473..8493)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /note="color: black; sequence: ACCCTAACTGACACACATTCC"
     promoter        8478..8807
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      8658..8793
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     8720..8739
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     promoter        8855..8902
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
                     /note="color: #ffffff; direction: RIGHT"
     CDS             8921..9295
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /note="color: #ccffcc"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    9425..9546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(9462..9481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     9516..9535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     primer_bind     complement(9595..9611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(9595..9611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     complement(9608..9630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     protein_bind    9619..9635
                     /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)."
                     /note="color: #31849b"
     promoter        complement(9643..9673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 9643 .. 9649 / #ffffff / -10
                        2: 9650 .. 9667 / #ffffff
                        3: 9668 .. 9673 / #ffffff / -35"
     protein_bind    9688..9709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     primer_bind     complement(9826..9843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     rep_origin      complement(9997..10585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(10077..10096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     CDS             complement(10756..11616)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 10756 .. 11547 / #ccffcc
                        2: 11548 .. 11616 / #ccffcc / signal sequence
                     Cleavage site after base 11547"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11379..11398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     promoter        complement(11617..11721)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(11796..11815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /note="color: black; sequence: CGGCATCAGAGCAGATTGTA"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaataactt cgtatagcat acattatacg aagttattaa
     2401 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactgga gtaacccacc
     2461 caagatctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc
     2521 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct
     2581 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2641 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2701 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     2761 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     2821 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     2881 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     2941 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3001 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3061 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3121 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg
     3181 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg
     3241 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc
     3301 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg
     3361 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc
     3421 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc
     3481 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc
     3541 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat
     3601 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt
     3661 ttttggcttt tttgttagac gaagcttggt accgagctcg gatctccacc ccgtaccggt
     3721 cctgcagtcg aattcaccat gtctagactg gacaagagca aagtcataaa cggagctctg
     3781 gaattactca atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     3841 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     3901 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     3961 tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc
     4021 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     4081 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     4141 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     4201 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     4261 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     4321 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat
     4381 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     4441 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaattaat taattaaccc
     4501 gtgtcggctc cagatcttga ggccctttcg tcttcactcg agtttactcc ctatcagtga
     4561 tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc agactttact
     4621 ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata gagaacgtat
     4681 gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc ctatcagtga
     4741 tagagaacgt atatccagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc
     4801 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     4861 caattccaca acacttttgt cttatactta cgcgtctgca ggatatcaag cgaagcttgg
     4921 gctgcaggtc gactctagag ccaccatggt gagcaagggc gaggaggtca tcaaagagtt
     4981 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
     5041 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5101 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
     5161 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
     5221 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     5281 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     5341 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     5401 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     5461 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     5521 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     5581 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag
     5641 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa
     5701 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     5761 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     5821 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     5881 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     5941 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     6001 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     6061 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     6121 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     6181 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     6241 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     6301 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga
     6361 cgagctgtac aagtagtaat agataacttc gtatagcata cattatacga agttatgaat
     6421 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     6481 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6541 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6601 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6661 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6721 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6781 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6841 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6901 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6961 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     7021 cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta
     7081 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     7141 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     7201 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     7261 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga
     7321 actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag
     7381 taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac
     7441 accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg
     7501 acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc
     7561 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7621 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7681 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
     7741 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     7801 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     7861 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     7921 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     7981 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     8041 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     8101 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     8161 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     8221 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     8281 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     8341 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     8401 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     8461 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     8521 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     8581 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     8641 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     8701 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     8761 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     8821 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     8881 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
     8941 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
     9001 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     9061 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     9121 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
     9181 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
     9241 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
     9301 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     9361 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     9421 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9541 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     9601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     9661 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     9721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     9781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     9841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    10561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    10621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    10681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    10741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    10801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    10861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    10921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    10981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    11041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    11101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    11161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    11221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    11281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    11341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    11401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    11461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    11521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    11581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    11641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    11701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    11761 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
    11821 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
    11881 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
    11941 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
