LOCUS       sequence-325286-        9609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentivirus sgRNA delivery backbone with Cre-removable
            tagBFP reporter and hygromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9609)
  AUTHORS   Dubrot J, Lane-Reticker SK, Kessler EA, Ayer A, Mishra G, Wolfe CH, 
            Zimmer MD, Du PP, Mahapatra A, Ockerman KM, Davis TGR, Kohnle IC, 
            Pope HW, Allen PM, Olander KE, Iracheta-Vellve A, Doench JG, Haining
            WN, Yates KB, Manguso RT
  TITLE     In vivo screens using a selective CRISPR antigen removal lentiviral 
            vector system reveal immune dependencies in renal cell carcinoma.
  JOURNAL   Immunity. 2021 Jan 20. pii: S1074-7613(21)00001-7. doi: 
            10.1016/j.immuni.2021.01.001.
  PUBMED    33497609
REFERENCE   2  (bases 1 to 9609)
  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: "Immunity. 
            2021 Jan 20. pii: S1074-7613(21)00001-7. doi: 
            10.1016/j.immuni.2021.01.001."
COMMENT     Sequence Label: pSCAR_sgRNA_hygro-tagBFP-lox2272
FEATURES             Location/Qualifiers
     source          1..9609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      711..1299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1200..1219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1453..1470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1587..1608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1623..1628,1629..1646,1647..1653)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1661..1677
                     /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     1666..1688
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1685..1701
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1685..1701
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1720..1740
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1722..1740
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1768..1994
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1995..2175
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2222..2347
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2840..3073
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3258..3302
                     /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        3483..3731
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3483..3503
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3654..3673
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3753..3828
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3890..4007
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(4066..4099)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        4180..5358
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4411..5349
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5306..5326
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5374..6393
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     CDS             6418..6474
                     /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"
     protein_bind    complement(7180..7213)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    7229..7817
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7282..7302)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7700..7711)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7889..8122
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8200..8321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8237..8256)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8291..8310
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8361..8496
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8516..8535)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8517..8535)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8545..8562)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8545..8561)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8554..8576)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8703..9158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8790..8809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9000..9021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9184..9288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9289..9357,9358..9609,1..540)
                     /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(9507..9526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
       61 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      121 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      181 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
      241 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      301 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      361 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      421 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
      481 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
      541 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
      601 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
      661 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      721 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      841 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1081 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1201 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1261 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1321 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1381 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1441 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     1501 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1561 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1621 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1681 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     1741 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     1801 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     1861 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     1921 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     1981 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2041 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2101 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2161 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     2221 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     2281 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     2341 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     2401 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     2461 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     2521 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     2581 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     2641 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     2701 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     2761 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     2821 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     2881 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     2941 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     3001 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     3061 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     3121 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     3181 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     3241 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     3301 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     3361 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     3421 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccggta
     3481 ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     3541 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     3601 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     3661 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     3721 gacgaaacac cggagacgct tttttcgtct cagtttgaga gctagaaata gcaagttcaa
     3781 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc
     3841 aagcttggcg taactagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa
     3901 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     3961 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     4021 gggacagcag agatccactt tggcgccggc tcgagggggc ccgggataac ttcgtatagg
     4081 atactttata cgaagttatt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     4141 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     4201 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
     4261 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4321 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4381 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     4441 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     4501 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     4561 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     4621 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     4681 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     4741 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     4801 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     4861 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     4921 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     4981 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     5041 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     5101 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     5161 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     5221 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     5281 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     5341 ccatttcagg tgtcgtgacg tacggccacc accatgaaga agcctgagct gaccgccaca
     5401 tccgtggaga agttcctgat cgagaagttt gactctgtga gcgatctgat gcagctgtct
     5461 gagggcgagg agagcagggc cttcagcttc gacgtgggcg gaaggggcta cgtgctgaga
     5521 gtgaacagct gcgccgacgg attctacaag gatagatacg tgtaccgcca ctttgcttcc
     5581 gccgctctgc caatccctga ggtgctggac atcggcgagt tctccgagtc tctgacctac
     5641 tgtatcagca ggagagccca gggagtgacc ctgcaggatc tgcctgagac agagctgcca
     5701 gccgtgctgc agccagtggc tgaggctatg gacgctatcg ccgctgccga tctgtctcag
     5761 acaagcggct tcggaccctt tggccctcag ggcatcggac agtacaccac atggcgcgac
     5821 ttcatctgcg ctatcgccga tccccacgtg taccactggc agaccgtgat ggacgataca
     5881 gtgagcgcct ccgtggctca ggccctggac gagctgatgc tgtgggccga ggattgtcct
     5941 gaggtgcggc acctggtgca cgctgacttc ggctccaaca acgtgctgac cgataacgga
     6001 cgcatcacag ccgtgatcga ctggagcgag gctatgtttg gcgattccca gtacgaggtg
     6061 gccaacatct tcttttggag gccatggctg gcttgcatgg agcagcagac cagatacttc
     6121 gagcggcgcc accctgagct ggctggatct ccaaggctga gagcttacat gctgaggatc
     6181 ggactggacc agctgtacca gagcctggtg gatggcaact ttgacgatgc tgcttgggct
     6241 cagggacggt gcgacgctat cgtgcgctcc ggagctggaa ccgtgggaag aacacagatc
     6301 gccaggagat ctgctgccgt gtggacagac ggatgcgtgg aggtgctggc tgattctgga
     6361 aaccggcgcc caagcacccg gccccgcgct aaggacctgc agctgcaggg atccggagct
     6421 acaaacttct ctctgctgaa gcaggctggc gatgtggagg agaacccagg acccgccagc
     6481 tccgagctga tcaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac
     6541 caccacttta agtgcacatc tgagggcgag ggaaagccct acgagggaac ccagacaatg
     6601 cggatcaagg tggtggaggg cggacctctg ccattcgctt ttgacatcct ggccaccagc
     6661 ttcctgtacg gctccaagac ctttatcaac cacacacagg gaatcccaga cttctttaag
     6721 cagtccttcc ccgagggctt tacctgggag cgggtgacca catacgagga tggcggagtg
     6781 ctgaccgcca cacaggacac aagcctgcag gatggatgtc tgatctacaa cgtgaagatc
     6841 cgcggcgtga acttcacctc caacggacct gtgatgcaga agaagacact gggctgggag
     6901 gcctttaccg agacactgta cccagctgac ggcggactgg agggaaggaa cgatatggcc
     6961 ctgaagctgg tgggcggatc tcacctgatc gctaacatca agaccacata cagaagcaag
     7021 aagcccgcca agaacctgaa gatgcctggc gtgtactacg tggactacag gctggagaga
     7081 atcaaggagg ctaacaacga gacctacgtg gagcagcacg aggtggctgt ggccagatac
     7141 tgtgatctgc caagcaagct gggccacaag ctgaattaga taacttcgta taggatactt
     7201 tatacgaagt tatacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     7261 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     7321 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     7381 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     7441 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     7501 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     7561 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     7621 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     7681 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     7741 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     7801 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     7861 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     7921 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     7981 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     8041 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     8101 tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat
     8161 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     8221 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8281 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     8341 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     8401 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     8461 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     8521 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     8581 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     8641 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     8701 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     8761 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     8821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8881 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     8941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     9001 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     9061 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     9121 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     9181 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     9241 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     9301 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     9361 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     9421 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     9481 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     9541 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     9601 ccagtcaca
//
