LOCUS       sequence-334408-       11255 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector to co-express an spsgRNA with NLS-mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11255)
  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 11255)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: 
            pW211-lenti-spsgRNA-Esp3I-2kb-filler-pEF1s-NLS-mNeonGreen-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..11255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..33)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        18..347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      198..333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     260..279
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        395..442
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             461..835
                     /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    965..1086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1002..1021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1056..1075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1135..1151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1135..1151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1148..1170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1159..1175
                     /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(1183..1189,1190..1207,1208..1213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1228..1249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1366..1383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1537..2125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1617..1636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2296..3087,3088..3156))
                     /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     2919..2938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3157..3261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3336..3355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3527..3906
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3908..4106
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4061..4081
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4124..4304
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4351..4476
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4969..5202
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5387..5431
                     /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    5729..5846
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5897..6145
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5897..5917
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6068..6087
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8027..8102
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8164..8375
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             8403..8450
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8451..9155
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      8703..8712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9165..9221
                     /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             9222..9620
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9636..10224
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9689..9709)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10107..10118)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10296..10529
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10555..10572)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10561..10785
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(10831..11255,1..4)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10918..10937)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11128..11149
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
       61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
      181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
      301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
      361 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
      421 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
      481 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
      541 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
      601 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
      661 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
      721 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
      781 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
      841 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
      901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
      961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     1141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     3301 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3361 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     3421 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     3481 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
     3541 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3601 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3661 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3781 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3841 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     3901 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     3961 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4021 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     4081 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
     4141 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     4201 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     4261 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     4321 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     4381 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     4441 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     4501 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     4561 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     4621 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     4681 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     4741 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     4801 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     4861 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     4921 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     4981 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     5041 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     5101 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     5161 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     5221 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     5281 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     5341 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     5401 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     5461 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     5521 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     5581 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     5641 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
     5701 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     5761 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     5821 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     5881 ggttaattaa ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata
     5941 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     6001 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     6061 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     6121 tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa tgagcacaca aaatacacat
     6181 gctaaaatat tatattctat gacctttata aaatcaacca aaatcttctt tttaataact
     6241 ttagtatcaa taattagaat ttttatgttc ctttttgcaa acttttaata aaaatgagca
     6301 aaataaaaaa acgctagttt tagtaactcg cgttgttttc ttcaccttta ataatagcta
     6361 ctccaccact tgttcctaag cggtcagctc ctgcttcaat cattttttga gcatcttcaa
     6421 atgttctaac tccaccagct gctttaacta aagcattgtc tttaacaact gacttcatta
     6481 gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt ttaacaaatt
     6541 ctaatccagc ttcaacagct atttcacaag ctttcatgat ttcttctttt gttaataaac
     6601 aattttccat aatacattta acaacatgtg atccagctgc tttttttaca gctttcatgt
     6661 cttctaaaac taattcataa tttttgtctt ttaatgcacc aatatttaat accatatcaa
     6721 tttctgttgc accatcttta attgcttcag aaacttcgaa tgcttttgta gctgttgtgc
     6781 atgcacctag aggaaaacct acaacatttg ttattcctac atttgtgcct tttaataatt
     6841 ctttacaata gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca aattcaattg
     6901 cttcatcaca taattgttta atttcagctt tcgtagcatc ttgttttaat aatgtgtgat
     6961 ctatatattt gtttagtttc attttttctc ctatatattc atttttaatt ttaattcttt
     7021 aataatttcg tctactttaa ctttagcgtt ttgaacagat tcaccaacac ctataaaata
     7081 aatttttagt ttaggttcag ttccacttgg gcgaacagca aatcatgact tatcttctaa
     7141 ataaaatttt agtaagtctt gtcctggcat attatacatt ccatcgatgt agtcttcaac
     7201 attaacaact ttaagtccag caatttgagt taagggtgtt gctctcaatg atttcattaa
     7261 tggttcaatt tttaatttct tttcttctgg tttaaaattc aagtttaaag tgaaagtgta
     7321 atatgcaccc atttctttaa ataaatcttc taaatagtct actaatgttt tattttgttt
     7381 tttataaaat caagcagcct ctgctattaa tatagaagct tgtattccat ctttatctct
     7441 agctgagtca tcaattacat atccataact ttcttcataa gcaaaaacaa aatttaatcc
     7501 gttatcttct tctttagcaa tttctctacc cattcattta aatccagtta aagtttttac
     7561 aatattaact ccatattttt catgagcgat tctatcaccc aaatcacttg ttacaaaact
     7621 tgaatataga gccggatttt ttggaatgct atttaagcgt tttagatttg ataattttca
     7681 atcaattaaa attggtcctg tttgatttcc atctaatctt acaaaatgac catcatgttt
     7741 tattgccatt ccaaatctgt cagcatctgg gtcattcata ataataatat ctgcatcatg
     7801 tttaatacca tattcaagcg gtatttttca tgcaggatca aattctggat ttggatttac
     7861 aacattttta aatgtttcat cttcaaatgc atgctcttca acctcaataa cgttatatcc
     7921 tgattcacgt aatatttttg gggtaaattt agttcctgtt ccattaactg cgctaaaaat
     7981 aatttttaaa tcttttttag cttcttgctc ttttttgtac gtctctgttt tagagctaga
     8041 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     8101 gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     8161 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     8221 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     8281 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     8341 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca
     8401 tgaagcgacc tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gtgagcaagg
     8461 gcgaggagga taacatggcc tctctcccag cgacacatga gttacacatc tttggctcca
     8521 tcaacggtgt ggactttgac atggtgggtc agggcaccgg caatccaaat gatggttatg
     8581 aggagttaaa cctgaagtcc accaagggtg acctccagtt ctccccctgg attctggtcc
     8641 ctcatatcgg gtatggcttc catcagtacc tgccctaccc tgacgggatg tcgcctttcc
     8701 aggccgccat ggtagatggc tccggatacc aagtccatcg cacaatgcag tttgaagatg
     8761 gtgcctccct tactgttaac taccgctaca cctacgaggg aagccacatc aaaggagagg
     8821 cccaggtgaa ggggactggt ttccctgctg acggtcctgt gatgaccaac tcgctgaccg
     8881 ctgcggactg gtgcaggtcg aagaagactt accccaacga caaaaccatc atcagtacct
     8941 ttaagtggag ttacaccact ggaaatggca agcgctaccg gagcactgcg cggaccacct
     9001 acacctttgc caagccaatg gcggctaact atctgaagaa ccagccgatg tacgtgttcc
     9061 gtaagacgga gctcaagcac tccaagaccg agctcaactt caaggagtgg caaaaggcct
     9121 ttaccgatgt gatgggcatg gacgagctgt acaagggatc cggcgcaaca aacttctctc
     9181 tgctgaaaca agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc
     9241 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     9301 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     9361 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     9421 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     9481 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     9541 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     9601 gttatgtgtg ggagggctaa acgcgttaag tcgacaatca acctctggat tacaaaattt
     9661 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     9721 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     9781 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     9841 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     9901 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     9961 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
    10021 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
    10081 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
    10141 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
    10201 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
    10261 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
    10321 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
    10381 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    10441 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    10501 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
    10561 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    10621 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    10681 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    10741 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    10801 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    10861 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    10921 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    10981 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    11041 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    11101 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    11161 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    11221 taaaaaatga gctgatttaa caaaaattta acgcg
//
