pWPI-N3ICD

LOCUS      pWPI-N3ICD      13103 bp DNA     circular SYN 25-AUG-2024
DEFINITION  Constitutive overexpression of human NOTCH3 intracellular domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13103)
  AUTHORS   Gastfriend BD, Snyder ME, Holt HE, Daneman R, Palecek SP, Shusta EV
  TITLE     Notch3 directs differentiation of brain mural cells from human 
            pluripotent stem cell-derived neural crest.
  JOURNAL   Sci Adv. 2024 Feb 2;10(5):eadi1737. doi: 10.1126/sciadv.adi1737. 
            Epub 2024 Feb 2.
  PUBMED    38306433
REFERENCE   2  (bases 1 to 13103)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2024 from SnapGene Server 7.0.3
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2024-02-2"; volume: "10(5):eadi1737. doi"; pages: " 
            10.1126/sciadv.adi1737. Epub 2024 Feb 2"
FEATURES             Location/Qualifiers
     source          1..13103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1299..1532
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1717..1761
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2144..3322
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2375..3313
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3270..3290
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3350..3359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5370..5487
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5536..6103
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5554..6105
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5698..5715)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5925..5944
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6108..6824
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             6108..6824
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6153..6174)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6414..6433)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6761..6782
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6903..6919)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6904..6920)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    6933..7521
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6986..7006)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7404..7415)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(7665..7698)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7718..7898
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7923..7941)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7924..7941)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8231..8250)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(8464..8483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8583..8605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8643..8661)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8729..8833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8834..9694
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9052..9071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9865..10453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10354..10373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10607..10624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10694..10714)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10699..11028
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10879..11014
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          12162..12227
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12357..12377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12802..12936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12839..12858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12893..12912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1201 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1261 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1321 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1381 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1441 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1501 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1561 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1621 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1681 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1741 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1801 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     1861 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     1921 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     1981 gacagatcca ttcgattagt gaacggatct cgacggtatc gatcacgaga ctagcctcga
     2041 ccatcgatgt cgacgataag ctttgcaaag atggataaag ttttaaacag agaggaatct
     2101 ttgcagctaa tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca
     2161 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     2221 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     2281 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     2341 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     2401 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     2461 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     2521 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     2581 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     2641 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     2701 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     2761 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     2821 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     2881 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     2941 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     3001 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     3061 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     3121 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     3181 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     3241 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     3301 tcttccattt caggtgtcgt gaggaatttc gacatttaaa tttaattaag ccaccatggt
     3361 catggtggcc cggcgcaagc gcgagcacag caccctctgg ttccctgagg gcttctcact
     3421 gcacaaggac gtggcctctg gtcacaaggg ccggcgggaa cccgtgggcc aggacgcgct
     3481 gggcatgaag aacatggcca agggtgagag cctgatgggg gaggtggcca cagactggat
     3541 ggacacagag tgcccagagg ccaagcggct aaaggtagag gagccaggca tgggggctga
     3601 ggaggctgtg gattgccgtc agtggactca acaccatctg gttgctgctg acatccgcgt
     3661 ggcaccagcc atggcactga caccaccaca gggcgacgca gatgctgatg gcatggatgt
     3721 caatgtgcgt ggcccagatg gcttcacccc gctaatgctg gcttccttct gtgggggggc
     3781 tctggagcca atgccaactg aagaggatga ggcagatgac acatcagcta gcatcatctc
     3841 cgacctgatc tgccaggggg ctcagcttgg ggcacggact gaccgtactg gcgagactgc
     3901 tttgcacctg gctgcccgtt atgcccgtgc tgatgcagcc aagcggctgc tggatgctgg
     3961 ggcagacacc aatgcccagg accactcagg ccgcactccc ctgcacacag ctgtcacagc
     4021 cgatgcccag ggtgtcttcc agattctcat ccgaaaccgc tctacagact tggatgcccg
     4081 catggcagat ggctcaacgg cactgatcct ggcggcccgc ctggcagtag agggcatggt
     4141 ggaagagctc atcgccagcc atgctgatgt caatgctgtg gatgagcttg ggaaatcagc
     4201 cttacactgg gctgcggctg tgaacaacgt ggaagccact ttggccctgc tcaaaaatgg
     4261 agccaataag gacatgcagg atagcaagga ggagaccccc ctattcctgg ccgcccgcga
     4321 gggcagctat gaggctgcca agctgctgtt ggaccacttt gccaaccgtg agatcaccga
     4381 ccacctggac aggctgccgc gggacgtagc ccaggagaga ctgcaccagg acatcgtgcg
     4441 cttgctggat caacccagtg ggccccgcag cccccccggt ccccacggcc tggggcctct
     4501 gctctgtcct ccaggggcct tcctccctgg cctcaaagcg gcacagtcgg ggtccaagaa
     4561 gagcaggagg ccccccggga aggcggggct ggggccgcag gggccccggg ggcggggcaa
     4621 gaagctgacg ctggcctgcc cgggccccct ggctgacagc tcggtcacgc tgtcgcccgt
     4681 ggactcgctg gactccccgc ggcctttcgg tgggccccct gcttcccctg gtggcttccc
     4741 ccttgagggg ccctatgcag ctgccactgc cactgcagtg tctctggcac agcttggtgg
     4801 cccaggccgg gcgggtctag ggcgccagcc ccctggagga tgtgtactca gcctgggcct
     4861 gctgaaccct gtggctgtgc ccctcgattg ggcccggctg cccccacctg cccctccagg
     4921 cccctcgttc ctgctgccac tggcgccggg accccagctg ctcaacccag ggacccccgt
     4981 ctccccgcag gagcggcccc cgccttacct ggcagtccca ggacatggcg aggagtaccc
     5041 ggcggctggg gcacacagca gccccccaaa ggcccgcttc ctgcgggttc ccagtgagca
     5101 cccttacctg accccatccc ccgaatcccc tgagcactgg gccagcccct cacctccctc
     5161 cctctcagac tggtccgaat ccacgcctag cccagccact gccactgggg ccatggccac
     5221 caccactggg gcactgcctg cccagccact tcccttgtct gttcccagct cccttgctca
     5281 ggcccagacc cagctggggc cccagccgga agttaccccc aagaggcaag tgttggcctg
     5341 attaattaat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac
     5401 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     5461 aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga ggtttaaact
     5521 acgggctgca ggaattccgc cccccccccc ctaacgttac tggccgaagc cgcttggaat
     5581 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     5641 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     5701 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     5761 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5821 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5881 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5941 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     6001 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc
     6061 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taataccatg gtgagcaagg
     6121 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     6181 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     6241 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     6301 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     6361 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     6421 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     6481 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     6541 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     6601 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     6661 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     6721 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     6781 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc
     6841 gactagctag tagctagcta gctagtcgag ctcaagcttc gaattcgata tcaagcttat
     6901 cgcgataccg tcgacctcga gggaattccg ataatcaacc tctggattac aaaatttgtg
     6961 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     7021 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     7081 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     7141 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     7201 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     7261 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     7321 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     7381 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     7441 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     7501 ccctttgggc cgcctccccg catcgggaat tcgagctcgg tacctttaag accaatgact
     7561 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     7621 attcactccc aacgaagaca agatgggatc aattcaccat gggaataact tcgtatagca
     7681 tacattatac gaagttatgc tgctttttgc ttgtactggg tctctctggt tagaccagat
     7741 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7801 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7861 cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atctagaatt aattccgtgt
     7921 attctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag
     7981 ccgcgttcta acgacaatat gtacaagcct aattgtgtag catctggctt actgaagcag
     8041 accctatcat ctctctcgta aactgccgtc agagtcggtt tggttggacg aaccttctga
     8101 gtttctggta acgccgtccc gcacccggaa atggtcagcg aaccaatcag cagggtcatc
     8161 gctagccaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     8221 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     8281 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
     8341 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
     8401 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaat ggtgcactct
     8461 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     8521 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     8581 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     8641 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     8701 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     8761 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     8821 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     8881 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     8941 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     9001 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     9061 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     9121 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     9181 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     9241 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     9301 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     9361 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     9421 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     9481 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     9541 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     9601 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     9661 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     9721 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     9781 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     9841 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     9901 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     9961 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
    10021 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
    10081 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
    10141 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
    10201 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
    10261 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
    10321 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
    10381 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
    10441 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
    10501 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
    10561 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
    10621 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
    10681 ttaatgcagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
    10741 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
    10801 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
    10861 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
    10921 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
    10981 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttggacac
    11041 aagacaggct tgcgagatat gtttgagaat accactttat cccgcgtcag ggagaggcag
    11101 tgcgtaaaaa gacgcggact catgtgaaat actggttttt agtgcgccag atctctataa
    11161 tctcgcgcaa cctattttcc cctcgaacac tttttaagcc gtagataaac aggctgggac
    11221 acttcacatg agcgaaaaat acatcgtcac ctgggacatg ttgcagatcc atgcacgtaa
    11281 actcgcaagc cgactgatgc cttctgaaca atggaaaggc attattgccg taagccgtgg
    11341 cggtctgtac cgggtgcgtt actggcgcgt gaactgggta ttcgtcatgt cgataccgtt
    11401 tgtatttcca gctacgatca cgacaaccag cgcgagctta aagtgctgaa acgcgcagaa
    11461 ggcgatggcg aaggcttcat cgttattgat gacctggtgg ataccggtgg tactgcggtt
    11521 gcgattcgtg aaatgtatcc aaaagcgcac tttgtcacca tcttcgcaaa accggctggt
    11581 cgtccgctgg ttgatgacta tgttgttgat atcccgcaag atacctggat tgaacagccg
    11641 tgggatatgg gcgtcgtatt cgtcccgcca atctccggtc gctaatcttt tcaacgcctg
    11701 gcactgccgg gcgttgttct ttttaacttc aggcgggtta caatagtttc cagtaagtat
    11761 tctggaggct gcatccatga cacaggcaaa cctgagcgaa accctgttca aaccccgctt
    11821 taaacatcct gaaacctcga cgctagtccg ccgctttaat cacggcgcac aaccgcctgt
    11881 gcagtcggcc cttgatggta aaaccatccc tcactggtat cgcatgatta accgtctgat
    11941 gtggatctgg cgcggcattg acccacgcga aatcctcgac gtccaggcac gtattgtgat
    12001 gagcgatgcc gaacgtaccg acgatgattt atacgatacg gtgattggct accgtggcgg
    12061 caactggatt tatgagtggg ccccggatct ttgtgaagga accttacttc tgtggtgtga
    12121 cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa
    12181 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg
    12241 gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag
    12301 aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa
    12361 aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga
    12421 gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa
    12481 aagctgcact gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc
    12541 ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg
    12601 ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta
    12661 ataaggaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt
    12721 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
    12781 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
    12841 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    12901 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
    12961 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
    13021 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    13081 atatgtacta caaacttagt agt
//