LOCUS       sequence-340545-       11243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector to co-express an sasgRNA with NLS-mScarlet-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11243)
  AUTHORS   Wang S, Matsumoto K, Lish SR, Cartagena-Rivera AX, Yamada KM
  TITLE     Budding epithelial morphogenesis driven by cell-matrix versus 
            cell-cell adhesion.
  JOURNAL   Cell. 2021 Jul 8;184(14):3702-3716.e30. doi: 
            10.1016/j.cell.2021.05.015. Epub 2021 Jun 15.
  PUBMED    34133940
REFERENCE   2  (bases 1 to 11243)
  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: 
            "10.1016/j.cell.2021.05.015"; journalName: "Cell"; date: "2021-07-8-
            8"; volume: "184"; issue: "14"; pages: "3702-3716.e30"
COMMENT     Sequence Label: 
            pW214-lenti-sasgRNA-Esp3I-2kb-filler-pEF1s-NLS-mScarlet-I-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..11243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        89..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     331..350
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        466..513
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             532..906
                     /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    1036..1157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1073..1092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1127..1146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1206..1222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1206..1222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1219..1241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1230..1246
                     /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(1254..1260,1261..1278,1279..1284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1299..1320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1437..1454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1608..2196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1688..1707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2367..3158,3159..3227))
                     /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     2990..3009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3228..3332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3407..3426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3598..3977
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3979..4177
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4132..4152
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4195..4375
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4422..4547
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5040..5273
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5458..5502
                     /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    5800..5917
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5968..6216
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5968..5988
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6139..6158
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8098..8173
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        8235..8446
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             8474..8521
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8522..9214
                     /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(8652..8670)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8920..8939
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9224..9280
                     /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             9281..9679
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9695..10283
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9748..9768)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10166..10177)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10355..10588
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10614..10631)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10620..10844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(10890..11243,1..75)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10977..10996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11187..11208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
       61 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
      481 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
      541 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
      601 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
      661 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
      721 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
      781 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
      841 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
      901 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
      961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1021 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1141 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     1201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3361 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     3421 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     3481 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     3541 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     3601 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     3661 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3721 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3781 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3841 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3901 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3961 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4021 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4081 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4141 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
     4201 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4261 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4321 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     4381 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     4441 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     4501 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     4561 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     4621 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     4681 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     4741 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     4801 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     4861 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     4921 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     4981 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     5041 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     5101 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     5161 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     5221 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     5281 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     5341 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     5401 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     5461 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     5521 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     5581 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     5641 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     5701 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     5761 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     5821 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     5881 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     5941 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     6001 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     6061 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     6121 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     6181 ttggctttat atatcttgtg gaaaggacga aacaccggag acggttgtaa atgagcacac
     6241 aaaatacaca tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct
     6301 ttttaataac tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat
     6361 aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt
     6421 aataatagct actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg
     6481 agcatcttca aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac
     6541 tgacttcatt agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt
     6601 tttaacaaat tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt
     6661 tgttaataaa caattttcca taatacattt aacaacatgt gatccagctg ctttttttac
     6721 agctttcatg tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa
     6781 taccatatca atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt
     6841 agctgttgtg catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc
     6901 ttttaataat tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc
     6961 aaattcaatt gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa
     7021 taatgtgtga tctatatatt tgtttagttt cattttttct cctatatatt catttttaat
     7081 tttaattctt taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca
     7141 cctataaaat aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac
     7201 ttatcttcta aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg
     7261 tagtcttcaa cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat
     7321 gatttcatta atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa
     7381 gtgaaagtgt aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt
     7441 ttattttgtt ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca
     7501 tctttatctc tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca
     7561 aaatttaatc cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt
     7621 aaagttttta caatattaac tccatatttt tcatgagcga ttctatcacc caaatcactt
     7681 gttacaaaac ttgaatatag agccggattt tttggaatgc tatttaagcg ttttagattt
     7741 gataattttc aatcaattaa aattggtcct gtttgatttc catctaatct tacaaaatga
     7801 ccatcatgtt ttattgccat tccaaatctg tcagcatctg ggtcattcat aataataata
     7861 tctgcatcat gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga
     7921 tttggattta caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata
     7981 acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact
     8041 gcgctaaaaa taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt
     8101 atagtactct ggaaacagaa tctactataa caaggcaaaa tgccgtgttt atctcgtcaa
     8161 cttgttggcg agattttttg aattcgctag ctaggtcttg aaaggagtgg gaattggctc
     8221 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     8281 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     8341 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     8401 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggttctagag
     8461 cgctgccacc atgaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
     8521 agtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg
     8581 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     8641 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat
     8701 cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg ccgacatccc
     8761 cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     8821 ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta
     8881 caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac
     8941 aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga
     9001 cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac
     9061 ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga
     9121 catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg
     9181 ccactccacc ggcggcatgg acgagctgta caagggatcc ggcgcaacaa acttctctct
     9241 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atggccaagc ctttgtctca
     9301 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     9361 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     9421 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     9481 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     9541 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     9601 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     9661 ttatgtgtgg gagggctaaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     9721 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9781 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9841 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9901 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9961 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
    10021 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
    10081 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
    10141 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
    10201 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
    10261 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
    10321 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
    10381 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
    10441 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    10501 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    10561 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac
    10621 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
    10681 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
    10741 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
    10801 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
    10861 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
    10921 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
    10981 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
    11041 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
    11101 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
    11161 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
    11221 ccctatctcg gtctattctt ttg
//
