LOCUS       Exported                9550 bp ds-DNA     circular SYN 09-MAR-2021
DEFINITION  Lentiviral vector for mammalian expression of doxycycline-inducible 
            Cas9 and constitutive expression of Blasticidin S deaminaase -T2A 
            -rtTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9550)
  AUTHORS   Habashy et al
  TITLE     CRISPR/Cas9-based system for loss-of-function screening
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9550)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 7, 2022 from SnapGene 5.2.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..9550
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    220..238
                     /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    256..274
                     /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    291..309
                     /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    327..345
                     /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    363..381
                     /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    398..416
                     /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    434..452
                     /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     472..491
                     /label=121720_TRE_seqfw
                     /note="color: black; sequence: tgtacggtgggaggcctata; added:
                     2020-12-17"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
                     /note="color: black; sequence: AGCTCGTTTAGTGAACCGTCAGATC"
     misc_feature    513..567
                     /label=030221_CSK_TRE_fw
                     /note="color: #ff99cc; direction: RIGHT"
     misc_feature    541..546
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             547..1896
                     /codon_start=1
                     /label=CSK
                     /note="color: #00ffff"
                     /translation="MSAIQAAWPSGTECIAKYNFHGTAEQDLPFCKGDVLTIVAVTKDP
                     NWYKAKNKVGREGIIPANYVQKREGVKAGTKLSLMPWFHGKITREQAERLLYPPETGLF
                     LVRESTNYPGDYTLCVSCDGKVEHYRIMYHASKLSIDEEVYFENLMQLVEHYTSDADGL
                     CTRLIKPKVMEGTVAAQDEFYRSGWALNMKELKLLQTIGKGEFGDVMLGDYRGNKVAVK
                     CIKNDATAQAFLAEASVMTQLRHSNLVQLLGVIVEEKGGLYIVTEYMAKGSLVDYLRSR
                     GRSVLGGDCLLKFSLDVCEAMEYLEGNNFVHRDLAARNVLVSEDNVAKVSDFGLTKEAS
                     STQDTGKLPVKWTAPEALREKKFSTKSDVWSFGILLWEIYSFGRVPYPRIPLKDVVPRV
                     EKGYKMDAPDGCPPAVYEVMKNCWHLDAAMRPSFLQLREQLEHIKTHELHL"
     misc_feature    788..809
                     /label=030221_CSK_seqfw_1
                     /note="color: #ff99cc; direction: RIGHT"
     misc_feature    1212..1233
                     /label=030221_CSK_seqfw_2
                     /note="color: #ff99cc; direction: RIGHT"
     misc_feature    complement(1876..1926)
                     /label=030221_CSK_P2A_rv
                     /note="color: #ff99cc; direction: LEFT"
     misc_feature    1909..1965
                     /label=P2A
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(1937..1957)
                     /label=030221_P2A_seqrv
                     /note="color: #ff99cc; direction: LEFT"
     misc_feature    1975..2682
                     /label=mCherry
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(2221..2242)
                     /label=073019_mCherry_earlyseqrv
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    2558..2579
                     /label=080219_mCherry_seqfw
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        2693..3203
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(2709..2727)
                     /label=121720_PGK_seqrv
                     /note="color: black; sequence: cagggctgccttggaaaag; added: 
                     2020-12-17"
     primer_bind     3122..3140
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
                     /note="color: black; sequence: GTGTTCCGCATTCTGCAAG"
     CDS             3213..3608
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /note="color: #ccffcc"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             3609..3662
                     /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."
                     /note="color: #cc99b2
                     Cleavage site after base 3659"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3663..4409
                     /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: 3663 .. 4280 / #993366 / rTetR
                        2: 4281 .. 4286 / #cc99b2
                        3: 4287 .. 4322 / #993366 / VP16 minimal transcriptio...
                        4: 4323 .. 4325 / #cc99b2
                        5: 4326 .. 4361 / #993366 / VP16 minimal transcriptio...
                        6: 4362 .. 4364 / #cc99b2
                        7: 4365 .. 4400 / #993366 / VP16 minimal transcriptio...
                        8: 4401 .. 4409 / #cc99b2"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(4449..4469)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(4867..4878)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4866"
                     /translation="IEGR"
     LTR             5083..5316
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    5388..5509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5425..5444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     5479..5498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     rep_origin      5549..5684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     5611..5630
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(5634..5645)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     promoter        5684..5755
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5756..6616
                     /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: 5756 .. 5824 / #ccffcc / signal sequence
                        2: 5825 .. 6616 / #ccffcc
                     Cleavage site after base 5824"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5974..5993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      6787..7375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     7276..7295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     7529..7546
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    7663..7684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        7699..7729
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 7699 .. 7704 / #ffffff / -35
                        2: 7705 .. 7722 / #ffffff
                        3: 7723 .. 7729 / #ffffff / -10"
     protein_bind    7737..7753
                     /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"
     primer_bind     7742..7764
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     7761..7777
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     7761..7777
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     7796..7816
                     /label=T3
                     /note="T3 promoter, forward primer"
                     /note="color: black; sequence: GCAATTAACCCTCACTAAAGG"
     promoter        7798..7816
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        7844..8070
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             8071..8251
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    8298..8423
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    8916..9149
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gccaccatgt cagcaataca ggccgcctgg ccatccggta cagaatgtat tgccaagtac
      601 aacttccacg gcactgccga gcaggacctg cccttctgca aaggagacgt gctcaccatt
      661 gtggccgtca ccaaggaccc caactggtac aaagccaaaa acaaggtggg ccgtgagggc
      721 atcatcccag ccaactacgt ccagaagcgg gagggcgtga aggcgggtac caaactcagc
      781 ctcatgcctt ggttccacgg caagatcaca cgggagcagg ctgagcggct tctgtacccg
      841 ccggagacag gcctgttcct ggtgcgggag agcaccaact accccggaga ctacacgctg
      901 tgcgtgagct gcgacggcaa ggtggagcac taccgcatca tgtaccatgc cagcaagctc
      961 agcatcgacg aggaggtgta ctttgagaac ctcatgcagc tggtggagca ctacacctca
     1021 gacgcagatg gactctgtac gcgcctcatt aaaccaaagg tcatggaggg cacagtggcg
     1081 gcccaggatg agttctaccg cagcggctgg gccctgaaca tgaaggagct gaagctgctg
     1141 cagaccatcg ggaaggggga gttcggagac gtgatgctgg gcgattaccg agggaacaaa
     1201 gtcgccgtca agtgcattaa gaacgacgcc actgcccagg ccttcctggc tgaagcctca
     1261 gtcatgacgc aactgcggca tagcaacctg gtgcagctcc tgggcgtgat cgtggaggag
     1321 aagggcgggc tctacatcgt cactgagtac atggccaagg ggagccttgt ggactacctg
     1381 cggtctaggg gtcggtcagt gctgggcgga gactgtctcc tcaagttctc gctagatgtc
     1441 tgcgaggcca tggaatacct ggagggcaac aatttcgtgc atcgagacct ggctgcccgc
     1501 aatgtgctgg tgtctgagga caacgtggcc aaggtcagcg actttggtct caccaaggag
     1561 gcgtccagca cccaggacac gggcaagctg ccagtcaagt ggacagcccc tgaggccctg
     1621 agagagaaga aattctccac taagtctgac gtgtggagtt tcggaatcct tctctgggaa
     1681 atctactcct ttgggcgagt gccttatcca agaattcccc tgaaggacgt cgtccctcgg
     1741 gtggagaagg gctacaagat ggatgccccc gacggctgcc cgcccgcagt ctatgaagtc
     1801 atgaagaact gctggcacct ggacgccgcc atgcggccct ccttcctaca gctccgagag
     1861 cagcttgagc acatcaaaac ccacgagctg cacctgaccg gtagcggagc tactaacttc
     1921 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctacgcg tatggtgagc
     1981 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     2041 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     2101 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     2161 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     2221 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     2281 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     2341 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     2401 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     2461 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     2521 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     2581 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     2641 ggccgccact ccaccggcgg catggacgag ctgtacaaga tcgtttaatt aaggggttgg
     2701 ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg
     2761 gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca
     2821 gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc
     2881 ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg
     2941 cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac
     3001 cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg
     3061 aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc
     3121 ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc
     3181 gaatcaccga cctctctccc cagcaattca ccatggccaa gcctttgtct caagaagaat
     3241 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     3301 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     3361 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     3421 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     3481 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     3541 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     3601 gggagggcga aggtagaggt tctctcctca cttgtggtga tgttgaagaa aaccctggtc
     3661 caatgtctag actggacaag agcaaagtca taaacggagc tctggaatta ctcaatggtg
     3721 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta
     3781 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc
     3841 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc
     3901 ggaacaacgc caagtcatac cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc
     3961 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc
     4021 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact
     4081 ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga
     4141 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc
     4201 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac
     4261 agctaaagtg cgaaagcggc gggccgaccg acgcccttga cgattttgac ttagacatgc
     4321 tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg
     4381 attttgacct tgacatgctc cccgggtaac taagtaagga tcgatccaag atatcgtatt
     4441 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4501 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4561 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4621 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4681 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4741 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     4801 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4861 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4921 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4981 ccgcctgttt cgcctcggcg tccggactag aggtaccttt aagaccaatg acttacaagg
     5041 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5101 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5161 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5221 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5281 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5341 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     5401 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5461 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     5521 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5581 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5641 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctttcaa atatgtatcc
     5701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5941 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6601 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6661 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6721 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7501 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7561 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7621 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7681 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     7741 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     7801 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     7861 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     7921 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     7981 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     8041 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     8101 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8161 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     8221 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     8281 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     8341 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     8401 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     8461 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     8521 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     8581 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     8641 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     8701 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     8761 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     8821 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     8881 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     8941 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     9001 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     9061 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     9121 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     9181 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     9241 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     9301 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     9361 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     9421 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     9481 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     9541 ccgaggggac
//
