LOCUS       sequence-331179-        9549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of negative control sgRNA 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_sgNeg1
FEATURES             Location/Qualifiers
     source          1..9549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..1250
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          303..1241
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1198..1218
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1257..1266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1272..2288
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK"
     CDS             2313..2369
                     /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(2370..2372,2373..2375,2376..3089)
                     /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(2370..2372,2373..2375,2376..3086)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2415..2436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2676..2695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3023..3044
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3104..3692
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3157..3177)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3575..3586)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3764..3997
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4075..4196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4112..4131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4166..4185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4236..4371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4391..4410)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4392..4410)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4420..4437)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4420..4436)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4429..4451)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4578..5033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4665..4684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4875..4896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5059..5163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5164..5232,5233..6024)
                     /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(5382..5401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6195..6783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6684..6703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6937..6954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7071..7092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7107..7112,7113..7130,7131..7137)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7145..7161
                     /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     7150..7172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7169..7185
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7169..7185
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7204..7224
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7206..7224
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7252..7478
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7479..7659
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7706..7831
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8324..8557
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8742..8786
                     /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        8964..9212
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8964..8984
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9135..9154
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9234..9309
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9365..9482
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag
       61 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
      121 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa
      181 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      241 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      301 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
      361 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
      421 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
      481 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
      541 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
      601 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
      661 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
      721 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
      781 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
      841 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
      901 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
      961 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     1021 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     1081 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     1141 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     1201 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga cgtacggcca
     1261 ccatggaatt caaaaagcct gaactcaccg ctacctctgt cgagaagttt ctgatcgaaa
     1321 agtttgacag tgtatccgac ctgatgcagc tctccgaggg cgaagaatct cgggctttca
     1381 gcttcgatgt gggagggcgt ggatatgtcc tgcgggtgaa tagctgcgcc gatggtttct
     1441 acaaagatcg ctatgtttat cggcactttg catccgccgc tctccctatt cccgaagtgc
     1501 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     1561 tcaccttgca agacctgcct gaaaccgaac tgcccgctgt tctccagccc gtcgccgagg
     1621 ccatggatgc catcgctgcc gccgatctta gccagaccag cgggttcggc ccattcggac
     1681 ctcaaggaat cggtcaatac actacatggc gcgatttcat ctgcgctatt gctgatcccc
     1741 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcctccgtc gcccaggctc
     1801 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgccg
     1861 atttcggctc caacaatgtc ctgaccgaca atggccgcat aacagccgtc attgactgga
     1921 gcgaggccat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccct
     1981 ggttggcttg tatggagcag cagacccgct acttcgagcg gaggcatccc gagcttgcag
     2041 gatctcctcg gctccgggct tatatgctcc gcattggtct tgaccaactc tatcagagct
     2101 tggttgacgg caatttcgat gatgcagctt gggctcaggg tcgctgcgac gcaatcgtcc
     2161 ggtccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgct gccgtctgga
     2221 ccgatggctg tgtggaagtg ctcgccgata gtggaaacag acgccccagc actcgtccta
     2281 gggcaaagga tctgcagtct agaggatctg gtgccacgaa tttctcacta ctcaagcagg
     2341 ccggtgatgt cgaggaaaac cctggtccta tggtgagcaa gggcgaggag ctgttcaccg
     2401 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2461 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2521 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2581 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2641 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2701 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2761 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2821 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2881 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2941 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3001 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3061 ctctcggcat ggacgagctg tacaagtgac gcgttaagtc gacaatcaac ctctggatta
     3121 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3181 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3241 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3301 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3361 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3421 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3481 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     3541 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3601 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3661 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     3721 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     3781 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     3841 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     3901 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     3961 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     4021 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     4081 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4141 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4201 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4321 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     4381 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     4441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4561 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4621 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4681 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4741 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4801 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4861 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4921 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4981 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5401 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5701 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6961 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7021 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7081 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7141 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7201 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     7261 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     7321 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     7381 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     7441 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     7501 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7561 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7621 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     7681 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     7741 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     7801 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     7861 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     7921 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     7981 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     8041 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     8101 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     8161 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     8221 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     8281 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     8341 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     8401 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     8461 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     8521 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     8581 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     8641 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     8701 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     8761 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     8821 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     8881 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     8941 tcccaacccc gaggggaccc agagagggcc tatttcccat gattccttca tatttgcata
     9001 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     9061 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     9121 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     9181 ctttatatat cttgtggaaa ggacgaaaca ccgcgccaaa cgtgccctga cgggttttag
     9241 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     9301 agtcggtgct tttttaagct tggcgtaact agatcttgag acaaatggca gtattcatcc
     9361 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     9421 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     9481 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag ggggcccggg
     9541 tgcaaagat
//
