LOCUS       sequence-331184-        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_sgAR10
FEATURES             Location/Qualifiers
     source          1..9549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        80..1258
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          311..1249
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1206..1226
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1265..1274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1280..2296
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     CDS             2321..2377
                     /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(2378..2380,2381..2383,2384..3097)
                     /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(2378..2380,2381..2383,2384..3094)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2423..2444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2684..2703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3031..3052
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3112..3700
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3165..3185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3583..3594)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3772..4005
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4083..4204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4120..4139)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4174..4193
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4244..4379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4399..4418)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4400..4418)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4428..4445)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4428..4444)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4437..4459)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4586..5041
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4673..4692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4883..4904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5067..5171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5172..5240,5241..6032)
                     /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(5390..5409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6203..6791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6692..6711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6945..6962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7079..7100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7115..7120,7121..7138,7139..7145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7153..7169
                     /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     7158..7180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7177..7193
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7177..7193
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7212..7232
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7214..7232
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7260..7486
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7487..7667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7714..7839
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8332..8565
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8750..8794
                     /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        8972..9220
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8972..8992
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9143..9162
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9242..9317
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9373..9490
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt
       61 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
      121 agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg
      181 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
      241 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
      301 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
      361 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct
      421 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg
      481 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct
      541 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc
      601 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg
      661 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc
      721 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg
      781 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
      841 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
      901 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
      961 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
     1021 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
     1081 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
     1141 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
     1201 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgacg
     1261 tacggccacc atggaattca aaaagcctga actcaccgct acctctgtcg agaagtttct
     1321 gatcgaaaag tttgacagtg tatccgacct gatgcagctc tccgagggcg aagaatctcg
     1381 ggctttcagc ttcgatgtgg gagggcgtgg atatgtcctg cgggtgaata gctgcgccga
     1441 tggtttctac aaagatcgct atgtttatcg gcactttgca tccgccgctc tccctattcc
     1501 cgaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     1561 acagggtgtc accttgcaag acctgcctga aaccgaactg cccgctgttc tccagcccgt
     1621 cgccgaggcc atggatgcca tcgctgccgc cgatcttagc cagaccagcg ggttcggccc
     1681 attcggacct caaggaatcg gtcaatacac tacatggcgc gatttcatct gcgctattgc
     1741 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cctccgtcgc
     1801 ccaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     1861 gcacgccgat ttcggctcca acaatgtcct gaccgacaat ggccgcataa cagccgtcat
     1921 tgactggagc gaggccatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     1981 gaggccctgg ttggcttgta tggagcagca gacccgctac ttcgagcgga ggcatcccga
     2041 gcttgcagga tctcctcggc tccgggctta tatgctccgc attggtcttg accaactcta
     2101 tcagagcttg gttgacggca atttcgatga tgcagcttgg gctcagggtc gctgcgacgc
     2161 aatcgtccgg tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgctgc
     2221 cgtctggacc gatggctgtg tggaagtgct cgccgatagt ggaaacagac gccccagcac
     2281 tcgtcctagg gcaaaggatc tgcagtctag aggatctggt gccacgaatt tctcactact
     2341 caagcaggcc ggtgatgtcg aggaaaaccc tggtcctatg gtgagcaagg gcgaggagct
     2401 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2461 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2521 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2581 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2641 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2701 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2761 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2821 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2881 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2941 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     3001 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3061 cgggatcact ctcggcatgg acgagctgta caagtgacgc gttaagtcga caatcaacct
     3121 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3181 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3241 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3301 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3361 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3421 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3481 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     3541 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3601 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3661 cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga
     3721 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     3781 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     3841 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     3901 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     3961 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt
     4021 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     4081 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4141 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4201 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4321 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4381 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     4441 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4501 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4561 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4621 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4681 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4741 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4801 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4861 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4921 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4981 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5041 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5401 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5581 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6001 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6061 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6121 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6961 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7021 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7081 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7141 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     7201 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     7261 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     7321 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     7381 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     7441 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     7501 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     7561 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     7621 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     7681 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     7741 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     7801 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     7861 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     7921 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     7981 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     8041 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     8101 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     8161 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     8221 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     8281 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     8341 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     8401 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     8461 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     8521 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     8581 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     8641 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     8701 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     8761 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     8821 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     8881 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     8941 gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga ttccttcata
     9001 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
     9061 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     9121 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     9181 tttcttggct ttatatatct tgtggaaagg acgaaacacc gtctgtctct taatacttga
     9241 agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     9301 tggcaccgag tcggtgcttt tttaagcttg gcgtaactag atcttgagac aaatggcagt
     9361 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     9421 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     9481 tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgaggg
     9541 ggcccgggt
//
