LOCUS       sequence-331182-        9549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of sgRNA targeted to the androgen receptor 
            (AR) promoter for CRISPRi knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Exp 
            Med."; date: "2021-02-1"; volume: "218(2). pii: 211509. doi"; pages:
            " 10.1084/jem.20201137"
COMMENT     Sequence Label: Lenti_CRISPRi_sgAR2
FEATURES             Location/Qualifiers
     source          1..9549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        64..1242
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          295..1233
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1190..1210
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1249..1258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1264..2280
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     CDS             2305..2361
                     /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             join(2362..2364,2365..2367,2368..3081)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2362..2364,2365..2367,2368..3078)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2407..2428)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2668..2687)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3015..3036
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3096..3684
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3149..3169)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3567..3578)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3756..3989
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4067..4188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4104..4123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4158..4177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4228..4363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4383..4402)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4384..4402)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4412..4429)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4412..4428)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4421..4443)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4570..5025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4657..4676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4867..4888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5051..5155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5156..5224,5225..6016)
                     /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(5374..5393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6187..6775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6676..6695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6929..6946
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7063..7084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7099..7104,7105..7122,7123..7129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7137..7153
                     /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     7142..7164
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7161..7177
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7161..7177
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7196..7216
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7198..7216
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7244..7470
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7471..7651
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7698..7823
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8316..8549
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8734..8778
                     /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        8956..9204
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8956..8976
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9127..9146
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9226..9301
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9357..9474
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
       61 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
      121 gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa
      181 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
      241 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
      301 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
      361 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      421 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      481 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      541 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      601 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      661 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      721 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      781 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      841 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      901 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      961 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     1021 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1081 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1141 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1201 acagtggttc aaagtttttt tcttccattt caggtgtcgt gacgtacggc caccatggaa
     1261 ttcaaaaagc ctgaactcac cgctacctct gtcgagaagt ttctgatcga aaagtttgac
     1321 agtgtatccg acctgatgca gctctccgag ggcgaagaat ctcgggcttt cagcttcgat
     1381 gtgggagggc gtggatatgt cctgcgggtg aatagctgcg ccgatggttt ctacaaagat
     1441 cgctatgttt atcggcactt tgcatccgcc gctctcccta ttcccgaagt gcttgacatt
     1501 ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcaccttg
     1561 caagacctgc ctgaaaccga actgcccgct gttctccagc ccgtcgccga ggccatggat
     1621 gccatcgctg ccgccgatct tagccagacc agcgggttcg gcccattcgg acctcaagga
     1681 atcggtcaat acactacatg gcgcgatttc atctgcgcta ttgctgatcc ccatgtgtat
     1741 cactggcaaa ctgtgatgga cgacaccgtc agtgcctccg tcgcccaggc tctcgatgag
     1801 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc cgatttcggc
     1861 tccaacaatg tcctgaccga caatggccgc ataacagccg tcattgactg gagcgaggcc
     1921 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc ctggttggct
     1981 tgtatggagc agcagacccg ctacttcgag cggaggcatc ccgagcttgc aggatctcct
     2041 cggctccggg cttatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     2101 ggcaatttcg atgatgcagc ttgggctcag ggtcgctgcg acgcaatcgt ccggtccgga
     2161 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ctgccgtctg gaccgatggc
     2221 tgtgtggaag tgctcgccga tagtggaaac agacgcccca gcactcgtcc tagggcaaag
     2281 gatctgcagt ctagaggatc tggtgccacg aatttctcac tactcaagca ggccggtgat
     2341 gtcgaggaaa accctggtcc tatggtgagc aagggcgagg agctgttcac cggggtggtg
     2401 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2461 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2521 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2581 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2641 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2701 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2761 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2821 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2881 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2941 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3001 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3061 atggacgagc tgtacaagtg acgcgttaag tcgacaatca acctctggat tacaaaattt
     3121 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3181 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3241 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3301 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3361 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3421 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3481 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3541 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3601 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3661 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     3721 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     3781 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     3841 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3901 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3961 cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat
     4021 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     4081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     4201 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4261 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4321 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     4381 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     4441 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4501 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4561 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4621 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4681 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4741 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4801 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4861 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4921 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4981 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     5041 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5101 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5161 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5221 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5281 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5341 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5401 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5461 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5521 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5581 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5641 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5701 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5761 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5821 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5881 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5941 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6001 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6061 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6121 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6181 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6241 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6301 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6361 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6421 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6481 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6541 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6601 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6661 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6721 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6781 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6841 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6901 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6961 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7021 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7081 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     7141 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     7201 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     7261 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     7321 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     7381 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     7441 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     7501 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7561 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7621 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     7681 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     7741 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     7801 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     7861 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     7921 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     7981 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     8041 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     8101 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     8161 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     8221 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     8281 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     8341 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     8401 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     8461 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     8521 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     8581 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     8641 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     8701 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     8761 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     8821 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     8881 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     8941 ccgaggggac ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac
     9001 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
     9061 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     9121 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     9181 atcttgtgga aaggacgaaa caccgcagcc taaccaggcg ggtcggtttt agagctagaa
     9241 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     9301 cttttttaag cttggcgtaa ctagatcttg agacaaatgg cagtattcat ccacaatttt
     9361 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     9421 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     9481 tattacaggg acagcagaga tccactttgg cgccggctcg agggggcccg ggtgcaaaga
     9541 tggataaag
//
