LOCUS       sequence-376602-        9383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector to co-express a mouse Itgb1 spsgRNA (sg1-Itgb1) 
            with NLS-mScarlet-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9383)
  AUTHORS   Sekiguchi R, Mehlferber MM, Matsumoto K, Wang S
  TITLE     Efficient Gene Knockout in Salivary Gland Epithelial Explant 
            Cultures.
  JOURNAL   J Dent Res. 2023 Feb;102(2):197-206. doi: 10.1177/00220345221128201.
            Epub 2022 Nov 10.
  PUBMED    36366748
REFERENCE   2  (bases 1 to 9383)
  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"; doi: 
            "10.1177/00220345221128201"; journalName: "J Dent Res"; date: 
            "2023-02"; volume: "102"; issue: "2"; pages: "197-206"
COMMENT     Sequence Label: 
            pW401-lenti-sg1-mmItgb1-pEF1s-NLS-mScarlet-I-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..9383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..221
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     176..196
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             239..419
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    466..591
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1084..1317
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1502..1546
                     /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"
     misc_feature    1844..1961
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2012..2260
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2012..2032
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2183..2202
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2282..2357
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2419..2630
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2658..2705
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2706..3398
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2836..2854)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3104..3123
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3408..3464
                     /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             3465..3863
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3879..4467
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3932..3952)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4350..4361)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4539..4772
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4798..4815)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4804..5028
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5074..5502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5161..5180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5371..5392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5511..5531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5516..5845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5696..5831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5758..5777
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5893..5940
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5959..6333
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6463..6584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6500..6519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6554..6573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6633..6649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6633..6649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6646..6668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6657..6673
                     /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)."
     promoter        complement(join(6681..6687,6688..6705,6706..6711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6726..6747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6864..6881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7035..7623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7115..7134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7794..8585,8586..8654))
                     /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     8417..8436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8655..8759)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8834..8853)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9025..9383,1..21)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
       61 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      121 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      181 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
      241 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      301 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      361 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      421 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
      481 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
      541 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
      601 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
      661 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
      721 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
      781 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
      841 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
      901 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
      961 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1021 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1081 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1141 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1201 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1261 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1321 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1381 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     1441 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     1501 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     1561 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     1621 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     1681 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     1741 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     1801 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     1861 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     1921 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     1981 gcagagatcc agtttggtta attaaggtac cgagggccta tttcccatga ttccttcata
     2041 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
     2101 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     2161 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     2221 tttcttggct ttatatatct tgtggaaagg acgaaacacc gcaaactgat caatccaatc
     2281 cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     2341 tggcaccgag tcggtgcttt tttgaattcg ctagctaggt cttgaaagga gtgggaattg
     2401 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     2461 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     2521 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     2581 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gaccggttct
     2641 agagcgctgc caccatgaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga
     2701 agaaagtgag caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     2761 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     2821 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg
     2881 acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac cccgccgaca
     2941 tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     3001 tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga
     3061 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     3121 agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg
     3181 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     3241 ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt
     3301 tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg
     3361 gccgccactc caccggcggc atggacgagc tgtacaaggg atccggcgca acaaacttct
     3421 ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgatggcc aagcctttgt
     3481 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     3541 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     3601 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     3661 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     3721 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     3781 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     3841 ctggttatgt gtgggagggc taaacgcgtt aagtcgacaa tcaacctctg gattacaaaa
     3901 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3961 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     4021 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     4081 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     4141 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     4201 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     4261 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     4321 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     4381 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     4441 ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc
     4501 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     4561 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     4621 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     4681 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     4741 acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct
     4801 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4861 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4921 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4981 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     5041 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     5101 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5161 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5221 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5281 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5341 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5401 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5461 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     5521 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5581 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5641 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5701 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5761 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5821 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5881 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     5941 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     6001 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     6061 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     6121 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     6181 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     6241 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     6301 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     6361 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6421 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6481 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6541 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6601 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6661 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6721 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6781 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6841 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6901 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7201 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7261 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7381 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7441 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8101 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8401 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8761 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8821 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8881 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8941 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     9001 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     9061 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     9121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     9181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     9241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     9301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     9361 ctacttggca gtacatctac gta
//
