LOCUS       sequence-334399-       11243 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector to co-express an spsgRNA 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 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: 
            pW212-lenti-spsgRNA-Esp3I-2kb-filler-pEF1s-NLS-mScarlet-I-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..11243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..103
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             122..496
                     /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    626..747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(663..682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     717..736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(796..812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(796..812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(809..831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    820..836
                     /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(844..850,851..868,869..874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    889..910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1027..1044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1198..1786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1278..1297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1957..2748,2749..2817))
                     /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     2580..2599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2818..2922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2997..3016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3188..3567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3569..3767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3722..3742
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3785..3965
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4012..4137
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4630..4863
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5048..5092
                     /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    5390..5507
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5558..5806
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5558..5578
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5729..5748
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7688..7763
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7825..8036
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             8064..8111
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8112..8804
                     /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(8242..8260)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8510..8529
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8814..8870
                     /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             8871..9269
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9285..9873
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9338..9358)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9756..9767)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9945..10178
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10204..10221)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10210..10434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10480..10908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10567..10586)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10777..10798
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10917..10937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(10922..11243,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11102..11237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11164..11183
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
       61 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
      121 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
      181 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
      241 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
      301 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
      361 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
      421 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
      481 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
      541 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
      601 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
      661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      721 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
      781 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
      841 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
      901 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      961 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1021 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1081 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2041 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2101 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2161 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2221 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2281 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2341 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2401 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2461 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2521 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2581 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2641 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2701 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2761 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2821 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2881 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2941 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     3001 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     3061 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     3121 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     3181 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     3241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3541 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3721 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg
     3781 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     3841 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     3901 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     3961 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     4021 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     4081 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     4141 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     4201 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     4261 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     4321 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     4381 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     4441 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     4501 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     4561 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     4621 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     4681 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     4741 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     4801 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     4861 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     4921 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     4981 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     5041 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     5101 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     5161 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     5221 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     5281 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     5341 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     5401 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     5461 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     5521 gggacagcag agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc
     5581 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
     5641 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     5701 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     5761 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggag acggttgtaa
     5821 atgagcacac aaaatacaca tgctaaaata ttatattcta tgacctttat aaaatcaacc
     5881 aaaatcttct ttttaataac tttagtatca ataattagaa tttttatgtt cctttttgca
     5941 aacttttaat aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt
     6001 cttcaccttt aataatagct actccaccac ttgttcctaa gcggtcagct cctgcttcaa
     6061 tcattttttg agcatcttca aatgttctaa ctccaccagc tgctttaact aaagcattgt
     6121 ctttaacaac tgacttcatt agtttaacat cttcaaatgt tgcacctgat tttgaaaatc
     6181 ctgttgatgt tttaacaaat tctaatccag cttcaacagc tatttcacaa gctttcatga
     6241 tttcttcttt tgttaataaa caattttcca taatacattt aacaacatgt gatccagctg
     6301 ctttttttac agctttcatg tcttctaaaa ctaattcata atttttgtct tttaatgcac
     6361 caatatttaa taccatatca atttctgttg caccatcttt aattgcttca gaaacttcga
     6421 atgcttttgt agctgttgtg catgcaccta gaggaaaacc tacaacattt gttattccta
     6481 catttgtgcc ttttaataat tctttacaat agcttgttca atatgaatta acacaaactg
     6541 ttgcaaaatc aaattcaatt gcttcatcac ataattgttt aatttcagct ttcgtagcat
     6601 cttgttttaa taatgtgtga tctatatatt tgtttagttt cattttttct cctatatatt
     6661 catttttaat tttaattctt taataatttc gtctacttta actttagcgt tttgaacaga
     6721 ttcaccaaca cctataaaat aaatttttag tttaggttca gttccacttg ggcgaacagc
     6781 aaatcatgac ttatcttcta aataaaattt tagtaagtct tgtcctggca tattatacat
     6841 tccatcgatg tagtcttcaa cattaacaac tttaagtcca gcaatttgag ttaagggtgt
     6901 tgctctcaat gatttcatta atggttcaat ttttaatttc ttttcttctg gtttaaaatt
     6961 caagtttaaa gtgaaagtgt aatatgcacc catttcttta aataaatctt ctaaatagtc
     7021 tactaatgtt ttattttgtt ttttataaaa tcaagcagcc tctgctatta atatagaagc
     7081 ttgtattcca tctttatctc tagctgagtc atcaattaca tatccataac tttcttcata
     7141 agcaaaaaca aaatttaatc cgttatcttc ttctttagca atttctctac ccattcattt
     7201 aaatccagtt aaagttttta caatattaac tccatatttt tcatgagcga ttctatcacc
     7261 caaatcactt gttacaaaac ttgaatatag agccggattt tttggaatgc tatttaagcg
     7321 ttttagattt gataattttc aatcaattaa aattggtcct gtttgatttc catctaatct
     7381 tacaaaatga ccatcatgtt ttattgccat tccaaatctg tcagcatctg ggtcattcat
     7441 aataataata tctgcatcat gtttaatacc atattcaagc ggtatttttc atgcaggatc
     7501 aaattctgga tttggattta caacattttt aaatgtttca tcttcaaatg catgctcttc
     7561 aacctcaata acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt
     7621 tccattaact gcgctaaaaa taatttttaa atctttttta gcttcttgct cttttttgta
     7681 cgtctctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     7741 aaaaagtggc accgagtcgg tgcttttttg aattcgctag ctaggtcttg aaaggagtgg
     7801 gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     7861 tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg
     7921 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     7981 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc
     8041 ggttctagag cgctgccacc atgaagcgac ctgccgccac aaagaaggct ggacaggcta
     8101 agaagaagaa agtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     8161 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     8221 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     8281 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg
     8341 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     8401 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     8461 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     8521 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     8581 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     8641 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     8701 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     8761 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggatcc ggcgcaacaa
     8821 acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atggccaagc
     8881 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     8941 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     9001 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     9061 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     9121 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     9181 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     9241 tgccctctgg ttatgtgtgg gagggctaaa cgcgttaagt cgacaatcaa cctctggatt
     9301 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     9361 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     9421 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     9481 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     9541 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     9601 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     9661 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     9721 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     9781 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     9841 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     9901 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     9961 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
    10021 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
    10081 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
    10141 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
    10201 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
    10261 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
    10321 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
    10381 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
    10441 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
    10501 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
    10561 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    10621 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    10681 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    10741 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
    10801 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    10861 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
    10921 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
    10981 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
    11041 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
    11101 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
    11161 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
    11221 aggctttttt ggaggcctag gct
//
