LOCUS       Exported                9232 bp ds-DNA     circular SYN 01-JUL-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 9232)
  AUTHORS   Habashy et al
  TITLE     CRISPR/Cas9-based system for loss-of-function screening
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9232)
  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            "Unpublished"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..9232
                     /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..570
                     /label=030221_RAB10_TRE_fw
                     /note="color: #ff99cc; direction: RIGHT"
     misc_feature    541..546
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             547..1146
                     /codon_start=1
                     /label=RAB10
                     /note="color: #00ffff"
                     /translation="MAKKTYDLLFKLLLIGDSGVGKTCVLFRFSDDAFNTTFISTIGID
                     FKIKTVELQGKKIKLQIWDTAGQERFHTITTSYYRGAMGIMLVYDITNGKSFENISKWL
                     RNIDEHANEDVERMLLGNKCDMDDKRVVPKGKGEQIAREHGIRFFETSAKANINIEKAF
                     LTLAEDILRKTPVKEPNSENVDISSGGGVTGWKSKCC"
     misc_feature    907..931
                     /label=030221_RAB10_seqfw_1
                     /note="color: #ff99cc; direction: RIGHT"
     CDS             1156..1209
                     /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 1206"
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    1210..1578
                     /label=RABIF
                     /note="color: #a6acb3"
     misc_feature    1591..1647
                     /label=P2A
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(1619..1639)
                     /label=030221_P2A_seqrv
                     /note="color: #ff99cc; direction: LEFT"
     misc_feature    1657..2364
                     /label=mCherry
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(1903..1924)
                     /label=073019_mCherry_earlyseqrv
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    2240..2261
                     /label=080219_mCherry_seqfw
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        2375..2885
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(2391..2409)
                     /label=121720_PGK_seqrv
                     /note="color: black; sequence: cagggctgccttggaaaag; added: 
                     2020-12-17"
     primer_bind     2804..2822
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
                     /note="color: black; sequence: GTGTTCCGCATTCTGCAAG"
     CDS             2895..3290
                     /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             3291..3344
                     /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 3341"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3345..4091
                     /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: 3345 .. 3962 / #993366 / rTetR
                        2: 3963 .. 3968 / #cc99b2
                        3: 3969 .. 4004 / #993366 / VP16 minimal transcriptio...
                        4: 4005 .. 4007 / #cc99b2
                        5: 4008 .. 4043 / #993366 / VP16 minimal transcriptio...
                        6: 4044 .. 4046 / #cc99b2
                        7: 4047 .. 4082 / #993366 / VP16 minimal transcriptio...
                        8: 4083 .. 4091 / #cc99b2"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(4131..4151)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(4549..4560)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4548"
                     /translation="IEGR"
     LTR             4765..4998
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    5070..5191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5107..5126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     5161..5180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     rep_origin      5231..5366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     5293..5312
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(5316..5327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     promoter        5366..5437
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5438..6298
                     /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: 5438 .. 5506 / #ccffcc / signal sequence
                        2: 5507 .. 6298 / #ccffcc
                     Cleavage site after base 5506"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5656..5675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      6469..7057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     6958..6977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     7211..7228
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    7345..7366
                     /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        7381..7411
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 7381 .. 7386 / #ffffff / -35
                        2: 7387 .. 7404 / #ffffff
                        3: 7405 .. 7411 / #ffffff / -10"
     protein_bind    7419..7435
                     /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     7424..7446
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     7443..7459
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     7443..7459
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     7478..7498
                     /label=T3
                     /note="T3 promoter, forward primer"
                     /note="color: black; sequence: GCAATTAACCCTCACTAAAGG"
     promoter        7480..7498
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        7526..7752
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             7753..7933
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    7980..8105
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    8598..8831
                     /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 gccaccatgg cgaagaagac gtacgacctg cttttcaagc tgctcctgat cggggattcc
      601 ggagtgggga agacctgcgt cctttttcgt ttttcggatg atgccttcaa tactaccttt
      661 atttccacca taggaataga cttcaagatc aaaacagttg aattacaagg aaagaagatc
      721 aagctacaga tatgggatac agcaggccag gagcgatttc acaccatcac aacctcctac
      781 tacagaggcg caatgggtat catgctagta tatgacatca ccaatggtaa aagttttgaa
      841 aacatcagca aatggcttag aaacatagat gagcatgcca atgaagatgt ggaaagaatg
      901 ttactaggaa acaagtgtga tatggacgac aaaagagttg tacctaaagg aaaaggagaa
      961 cagattgcaa gggagcatgg tattaggttt tttgagacta gtgcaaaagc aaatataaac
     1021 atcgaaaagg cgttcctcac gttagctgaa gatatccttc gaaagacccc tgtaaaagag
     1081 cccaacagtg aaaatgtaga tatcagcagt ggaggaggcg tgacaggctg gaagagcaaa
     1141 tgctgcggat ccggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat
     1201 cctggcccaa tggaaccagc ggagcagccg agcgagttag tgtcagccga gggccgaaac
     1261 cggaaggcgg tgctgtgcca gcgttgcggc tcccgggtgc tgcagccagg gaccgctctc
     1321 ttctctcgcc gacagctttt ccttccctcc atgagaaaga agccagctct gtctgacggc
     1381 agcaatcctg acggcgatct cctccaggaa cactggctgg ttgaggacat gttcattttt
     1441 gagaatgtgg gcttcaccaa ggacgtgggc aacatcaagt ttctggtctg cgcagactgt
     1501 gaaattggac caattggctg gcattgccta gatgacaaga acagtttcta tgtggccttg
     1561 gaacgagttt cccatgagac cggtagcgga gctactaact tcagcctgct gaagcaggct
     1621 ggagacgtgg aggagaaccc tggacctacg cgtatggtga gcaagggcga ggaggataac
     1681 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1741 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1801 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     1861 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     1921 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     1981 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2041 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2101 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2161 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2221 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2281 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2341 ggcatggacg agctgtacaa gatcgtttaa ttaaggggtt ggggttgcgc cttttccaag
     2401 gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg
     2461 gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg
     2521 ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt
     2581 ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc
     2641 tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca
     2701 atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg
     2761 cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca
     2821 agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc
     2881 cccagcaatt caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     2941 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     3001 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     3061 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     3121 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     3181 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     3241 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc gaaggtagag
     3301 gttctctcct cacttgtggt gatgttgaag aaaaccctgg tccaatgtct agactggaca
     3361 agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc gaaggcctga
     3421 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
     3481 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
     3541 acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
     3601 accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
     3661 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
     3721 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
     3781 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
     3841 tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
     3901 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
     3961 gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     4021 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     4081 tccccgggta actaagtaag gatcgatcca agatatcgta ttcttaacta tgttgctcct
     4141 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4201 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4261 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4321 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4381 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4441 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     4501 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4561 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4621 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgt ttcgcctcgg
     4681 cgtccggact agaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     4741 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     4801 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     4861 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     4921 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     4981 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     5041 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     5101 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5161 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     5221 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5281 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5341 gtagtgagga ggcttttttg gaggcctttc aaatatgtat ccgctcatga gacaataacc
     5401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7081 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7141 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7201 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7261 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     7321 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7381 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7441 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     7501 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
     7561 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
     7621 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
     7681 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
     7741 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7801 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7861 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7921 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     7981 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     8041 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     8101 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     8161 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     8221 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     8281 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     8341 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     8401 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     8461 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     8521 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     8581 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     8641 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     8701 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     8761 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     8821 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     8881 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     8941 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     9001 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     9061 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     9121 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     9181 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg ac
//
