LOCUS       Exported                9067 bp ds-DNA     circular SYN 08-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 9067)
  AUTHORS   Habashy et al
  TITLE     CRISPR/Cas9-based system for loss-of-function screening
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9067)
  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            "Unpublished"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..9067
                     /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    541..546
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     misc_feature    547..1413
                     /label=OTX2
                     /note="color: #a6acb3"
     misc_feature    735..754
                     /label=possible sgRNA with fewest mismatches in OTX1
                     /note="color: #a6acb3"
     misc_feature    1426..1482
                     /label=P2A
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1492..2199
                     /label=mCherry
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(1738..1759)
                     /label=073019_mCherry_earlyseqrv
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    2075..2096
                     /label=080219_mCherry_seqfw
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        2210..2720
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(2226..2244)
                     /label=121720_PGK_seqrv
                     /note="color: black; sequence: cagggctgccttggaaaag; added: 
                     2020-12-17"
     primer_bind     2639..2657
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
                     /note="color: black; sequence: GTGTTCCGCATTCTGCAAG"
     CDS             2730..3125
                     /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             3126..3179
                     /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 3176"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3180..3926
                     /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: 3180 .. 3797 / #993366 / rTetR
                        2: 3798 .. 3803 / #cc99b2
                        3: 3804 .. 3839 / #993366 / VP16 minimal transcriptio...
                        4: 3840 .. 3842 / #cc99b2
                        5: 3843 .. 3878 / #993366 / VP16 minimal transcriptio...
                        6: 3879 .. 3881 / #cc99b2
                        7: 3882 .. 3917 / #993366 / VP16 minimal transcriptio...
                        8: 3918 .. 3926 / #cc99b2"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(3966..3986)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     CDS             complement(4384..4395)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4383"
                     /translation="IEGR"
     LTR             4600..4833
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    4905..5026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4942..4961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     4996..5015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     rep_origin      5066..5201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     5128..5147
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(5151..5162)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     promoter        5201..5272
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5273..6133
                     /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: 5273 .. 5341 / #ccffcc / signal sequence
                        2: 5342 .. 6133 / #ccffcc
                     Cleavage site after base 5341"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5491..5510)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     rep_origin      6304..6892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     6793..6812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     7046..7063
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     protein_bind    7180..7201
                     /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        7216..7246
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 7216 .. 7221 / #ffffff / -35
                        2: 7222 .. 7239 / #ffffff
                        3: 7240 .. 7246 / #ffffff / -10"
     protein_bind    7254..7270
                     /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     7259..7281
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     7278..7294
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     7278..7294
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     7313..7333
                     /label=T3
                     /note="T3 promoter, forward primer"
                     /note="color: black; sequence: GCAATTAACCCTCACTAAAGG"
     promoter        7315..7333
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        7361..7587
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             7588..7768
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    7815..7940
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    8433..8666
                     /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 gccaccatga tgtcttatct taagcaaccg ccttacgcag tcaatgggct gagtctgacc
      601 acttcgggta tggacttgct gcacccctcc gtgggctacc cggccacccc ccggaaacag
      661 cgccgggaga ggacgacgtt cactcgggcg cagctagatg tgctggaagc actgtttgcc
      721 aagacccggt acccagacat cttcatgcga gaggaggtgg cactgaaaat caacttgccc
      781 gagtcgaggg tgcaggtatg gtttaagaat cgaagagcta agtgccgcca acaacagcaa
      841 caacagcaga atggaggtca aaacaaagtg agacctgcca aaaagaagac atctccagct
      901 cgggaagtga gttcagagag tggaacaagt ggccaattca ctcccccctc tagcacctca
      961 gtcccgacca ttgccagcag cagtgctcct gtgtctatct ggagcccagc ttccatctcc
     1021 ccactgtcag atcccttgtc cacctcctct tcctgcatgc agaggtccta tcccatgacc
     1081 tatactcagg cttcaggtta tagtcaagga tatgctggct caacttccta ctttgggggc
     1141 atggactgtg gatcatattt gacccctatg catcaccagc ttcccggacc aggggccaca
     1201 ctcagtccca tgggtaccaa tgcagtcacc agccatctca atcagtcccc agcttctctt
     1261 tccacccagg gatatggagc ttcaagcttg ggttttaact caaccactga ttgcttggat
     1321 tataaggacc aaactgcctc ctggaagctt aacttcaatg ctgactgctt ggattataaa
     1381 gatcagacat cctcgtggaa attccaggtt ttgaccggta gcggagctac taacttcagc
     1441 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctacgcgtat ggtgagcaag
     1501 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     1561 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1621 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     1681 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1741 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1801 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1861 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1921 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1981 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     2041 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     2101 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     2161 cgccactcca ccggcggcat ggacgagctg tacaagatcg tttaattaag gggttggggt
     2221 tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt
     2281 ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg
     2341 tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc
     2401 taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac
     2461 gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc
     2521 gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag
     2581 gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt
     2641 gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa
     2701 tcaccgacct ctctccccag caattcacca tggccaagcc tttgtctcaa gaagaatcca
     2761 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     2821 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     2881 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     2941 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     3001 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     3061 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     3121 agggcgaagg tagaggttct ctcctcactt gtggtgatgt tgaagaaaac cctggtccaa
     3181 tgtctagact ggacaagagc aaagtcataa acggagctct ggaattactc aatggtgtcg
     3241 gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc
     3301 tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg
     3361 acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac tttctgcgga
     3421 acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc
     3481 tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt
     3541 gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta
     3601 cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac
     3661 ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc gaccggcagg
     3721 gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc
     3781 taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta gacatgctcc
     3841 cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt
     3901 ttgaccttga catgctcccc gggtaactaa gtaaggatcg atccaagata tcgtattctt
     3961 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4021 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4081 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4141 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4201 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4261 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4321 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4381 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4441 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4501 cctgtttcgc ctcggcgtcc ggactagagg tacctttaag accaatgact tacaaggcag
     4561 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     4621 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     4681 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     4741 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     4801 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     4861 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     4921 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4981 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     5041 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5101 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5161 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctttcaaata tgtatccgct
     5221 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5281 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5341 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5401 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5461 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5521 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5581 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5641 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5701 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5761 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5821 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5881 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5941 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6001 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6061 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6121 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6961 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7021 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7201 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7261 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     7321 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     7381 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     7441 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     7501 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     7561 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     7621 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7681 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7741 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     7801 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     7861 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     7921 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     7981 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     8041 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     8101 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     8161 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     8221 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     8281 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     8341 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     8401 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     8461 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     8521 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     8581 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     8641 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     8701 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     8761 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     8821 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     8881 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     8941 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     9001 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     9061 aggggac
//
