LOCUS       Exported                9549 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Lentiviral expression of sgRNA targeted to the androgen receptor 
            (AR) promoter for CRISPRi knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    Lenti_CRISPRi_sgAR3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9549)
  AUTHORS   Ma M, Ghosh S, Tavernari D, Katarkar A, Clocchiatti A, Mazzeo L, 
            Samarkina A, Epiney J, Yu YR, Ho PC, Levesque MP, Ozdemir BC, 
            Ciriello G, Dummer R, Dotto GP
  TITLE     Sustained androgen receptor signaling is a determinant of melanoma 
            cell growth potential and tumorigenesis.
  JOURNAL   J Exp Med. 2021 Feb 1;218(2). pii: 211509. doi: 
            10.1084/jem.20201137.
  PUBMED    33112375
REFERENCE   2  (bases 1 to 9549)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9549
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        100..1278
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          331..1269
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1226..1246
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1285..1294
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1300..2316
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     CDS             2341..2397
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2398..3114
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2443..2464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2704..2723)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3051..3072
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3132..3720
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3185..3205)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3603..3614)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3792..4025
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4103..4224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4140..4159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4194..4213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4264..4399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4326..4345
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4419..4438)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4420..4438)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4448..4465)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4448..4464)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4457..4479)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4606..5061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4693..4712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4903..4924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5087..5191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5192..6052
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5410..5429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6223..6811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6712..6731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6965..6982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7099..7120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7135..7165
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7173..7189
                     /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)."
     primer_bind     7178..7200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7197..7213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7197..7213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7232..7252
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7234..7252
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7280..7506
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7507..7687
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7734..7859
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8352..8585
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8770..8814
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        8992..9232
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     8992..9012
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9163..9182
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9262..9337
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9393..9510
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cggctcgagg gggcccgggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
       61 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
      121 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
      181 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
      241 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
      301 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
      361 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
      421 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
      481 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
      541 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
      601 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
      661 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
      721 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
      781 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
      841 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
      901 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
      961 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     1021 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     1081 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     1141 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     1201 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     1261 ccatttcagg tgtcgtgacg tacggccacc atggaattca aaaagcctga actcaccgct
     1321 acctctgtcg agaagtttct gatcgaaaag tttgacagtg tatccgacct gatgcagctc
     1381 tccgagggcg aagaatctcg ggctttcagc ttcgatgtgg gagggcgtgg atatgtcctg
     1441 cgggtgaata gctgcgccga tggtttctac aaagatcgct atgtttatcg gcactttgca
     1501 tccgccgctc tccctattcc cgaagtgctt gacattgggg agttcagcga gagcctgacc
     1561 tattgcatct cccgccgtgc acagggtgtc accttgcaag acctgcctga aaccgaactg
     1621 cccgctgttc tccagcccgt cgccgaggcc atggatgcca tcgctgccgc cgatcttagc
     1681 cagaccagcg ggttcggccc attcggacct caaggaatcg gtcaatacac tacatggcgc
     1741 gatttcatct gcgctattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     1801 accgtcagtg cctccgtcgc ccaggctctc gatgagctga tgctttgggc cgaggactgc
     1861 cccgaagtcc ggcacctcgt gcacgccgat ttcggctcca acaatgtcct gaccgacaat
     1921 ggccgcataa cagccgtcat tgactggagc gaggccatgt tcggggattc ccaatacgag
     1981 gtcgccaaca tcttcttctg gaggccctgg ttggcttgta tggagcagca gacccgctac
     2041 ttcgagcgga ggcatcccga gcttgcagga tctcctcggc tccgggctta tatgctccgc
     2101 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     2161 gctcagggtc gctgcgacgc aatcgtccgg tccggagccg ggactgtcgg gcgtacacaa
     2221 atcgcccgca gaagcgctgc cgtctggacc gatggctgtg tggaagtgct cgccgatagt
     2281 ggaaacagac gccccagcac tcgtcctagg gcaaaggatc tgcagtctag aggatctggt
     2341 gccacgaatt tctcactact caagcaggcc ggtgatgtcg aggaaaaccc tggtcctatg
     2401 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2461 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2521 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2581 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2641 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2701 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2761 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2821 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2881 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2941 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3001 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3061 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgacgc
     3121 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3181 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3241 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3301 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3361 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3421 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3481 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3541 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3601 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3661 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3721 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     3781 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     3841 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     3901 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     3961 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     4021 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     4081 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     4141 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4201 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     4261 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4321 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4381 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     4441 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     4501 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     4561 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     4621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5041 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5401 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5701 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6961 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7021 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7081 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     7141 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7201 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     7261 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     7321 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     7381 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     7441 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     7501 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7561 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7621 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7681 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     7741 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     7801 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     7861 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     7921 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     7981 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     8041 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     8101 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     8161 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     8221 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     8281 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     8341 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     8401 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     8461 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     8521 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     8581 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     8641 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     8701 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     8761 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     8821 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     8881 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     8941 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta
     9001 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga
     9061 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     9121 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     9181 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
     9241 gtggacactg aatttggaag ggttttagag ctagaaatag caagttaaaa taaggctagt
     9301 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg gcgtaactag
     9361 atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     9421 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     9481 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     9541 ctttggcgc
//
