LOCUS       sequence-464744-       11862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a downstream of a ubiquitous chromatin opening element drives 
            expression of GOI and IRES GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11862)
  AUTHORS   Khan KA, Qureshi AA, Xu P, Kuo HY, Schumacher TN, Michael IP, Kerbel
            RS
  TITLE     Immunological Tolerance to Luciferase and Fluorescent Proteins using
            Tol Mice Enables Development of Improved Tumor Models for 
            Investigating Immunity and Metastasis.
  JOURNAL   Cancer Res. 2025 Apr 14. doi: 10.1158/0008-5472.CAN-24-2418.
  PUBMED    40228139
REFERENCE   2  (bases 1 to 11862)
  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"; journalName: "Cancer Res.
            2025 Apr 14. doi: 10.1158/0008-5472.CAN-24-2418."
COMMENT     Sequence Label: pLUEF1 IRES GFP (backbone)
FEATURES             Location/Qualifiers
     source          1..11862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     426..446
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    526..643
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    692..1259
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    710..1261
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(854..871)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1081..1100
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1264..1266,1267..1269,1270..1980)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1264..1266,1267..1269,1270..1980)
                     /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(1309..1330)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1570..1589)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1917..1938
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2059..2075)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2060..2076)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    2089..2677
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2142..2162)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2560..2571)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2821..2854)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             2874..3054
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3079..3097)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3080..3097)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3387..3406)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3620..3639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3739..3761
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3799..3817)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3885..3989
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3990..4058,4059..4850)
                     /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(4208..4227)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5021..5609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5510..5529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5763..5780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5850..5870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5855..6184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6035..6170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          7318..7383
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7513..7533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7958..8092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7995..8014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8049..8068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8260..8893
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8940..9065
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9560..9793
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9978..10022
                     /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    10307..10424
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(11156..11862,1..478)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(11387..11862,1..469)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
       61 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
      121 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
      181 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
      241 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
      301 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
      361 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
      421 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg
      481 aatttcgaca tttaaattta attaatctcg acggtatcgg ttaactttta aaagaaaagg
      541 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
      601 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttatcgatc acgagactag
      661 cctcgaggtt taaactacgg gctgcaggaa ttccgccccc ccccccctaa cgttactggc
      721 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
      781 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
      841 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
      901 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
      961 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1021 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     1081 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     1141 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     1201 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     1261 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1321 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1381 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1441 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1501 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1561 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1621 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1681 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1741 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1801 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1861 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1921 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1981 tccggactca gatctcgact agctagtagc tagctagcta gtcgagctca agcttcgaat
     2041 tcgatatcaa gcttatcgcg ataccgtcga cctcgaggga attccgataa tcaacctctg
     2101 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2161 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2221 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2281 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2341 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2401 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2461 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     2521 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2581 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2641 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gggaattcga gctcggtacc
     2701 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     2761 gggactggaa gggctaattc actcccaacg aagacaagat gggatcaatt caccatggga
     2821 ataacttcgt atagcataca ttatacgaag ttatgctgct ttttgcttgt actgggtctc
     2881 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     2941 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     3001 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct
     3061 agaattaatt ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt
     3121 tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc
     3181 tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt
     3241 tggacgaacc ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc
     3301 aatcagcagg gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac
     3361 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     3421 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     3481 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     3541 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     3601 tcgaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3661 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3721 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3781 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3841 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3901 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3961 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4021 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4081 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4141 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4201 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4261 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4321 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4381 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4441 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4501 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4561 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4621 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4681 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4741 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4801 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4861 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4921 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5161 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5581 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5641 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5701 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5761 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5821 cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6001 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6061 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6121 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6181 caaaaagctt ggacacaaga caggcttgcg agatatgttt gagaatacca ctttatcccg
     6241 cgtcagggag aggcagtgcg taaaaagacg cggactcatg tgaaatactg gtttttagtg
     6301 cgccagatct ctataatctc gcgcaaccta ttttcccctc gaacactttt taagccgtag
     6361 ataaacaggc tgggacactt cacatgagcg aaaaatacat cgtcacctgg gacatgttgc
     6421 agatccatgc acgtaaactc gcaagccgac tgatgccttc tgaacaatgg aaaggcatta
     6481 ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac tgggtattcg
     6541 tcatgtcgat accgtttgta tttccagcta cgatcacgac aaccagcgcg agcttaaagt
     6601 gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt attgatgacc tggtggatac
     6661 cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg tcaccatctt
     6721 cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt gttgatatcc cgcaagatac
     6781 ctggattgaa cagccgtggg atatgggcgt cgtattcgtc ccgccaatct ccggtcgcta
     6841 atcttttcaa cgcctggcac tgccgggcgt tgttcttttt aacttcaggc gggttacaat
     6901 agtttccagt aagtattctg gaggctgcat ccatgacaca ggcaaacctg agcgaaaccc
     6961 tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct agtccgccgc tttaatcacg
     7021 gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac catccctcac tggtatcgca
     7081 tgattaaccg tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc ctcgacgtcc
     7141 aggcacgtat tgtgatgagc gatgccgaac gtaccgacga tgatttatac gatacggtga
     7201 ttggctaccg tggcggcaac tggatttatg agtgggcccc ggatctttgt gaaggaacct
     7261 tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta
     7321 aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt
     7381 tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa
     7441 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa
     7501 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa
     7561 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt
     7621 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta
     7681 acctttataa gtaggcataa cagttataat cataacatac tgttttttct tactccacac
     7741 aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta
     7801 atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat
     7861 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     7921 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     7981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     8041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     8101 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     8161 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     8221 aaattgtatt tgttaaatat gtactacaaa cttagtagtt ggaagggcta attcactccc
     8281 aaagaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     8341 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
     8401 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
     8461 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
     8521 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
     8581 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
     8641 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
     8701 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     8761 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     8821 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     8881 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     8941 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     9001 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     9061 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     9121 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     9181 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     9241 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     9301 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     9361 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcgcggccg ctgatcttca
     9421 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     9481 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     9541 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     9601 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     9661 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     9721 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     9781 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     9841 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     9901 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     9961 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
    10021 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
    10081 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
    10141 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
    10201 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
    10261 tgaacggatc tcgacggtat cgccaaatgg cagtattcat ccacaatttt aaaagaaaag
    10321 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
    10381 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
    10441 acagcagaga tccagtttgg atcgataagc ttgatatcga attcgggagg tggtccctgc
    10501 agttacgcca atgataaccc ccgccagaaa aatcttagta gccttccctt tttgttttcc
    10561 gtgccccaac tcggcggatt gactcggccc cttccggaaa cacccgaatc aacttctagt
    10621 caaattattg ttcacgccgc aatgacccac ccctggcccg cgtctgtgga actgacccct
    10681 ggtgtacagg agagttcgct gctgaaagtg gtcccaaagg ggtactagtt tttaagctcc
    10741 caactccccc tcccccagcg tctggaggat tccacaccct cgcaccgcag gggcgaggaa
    10801 gtgggcggag tccggttttg gcgccagccg ctgaggctgc caagcagaaa agccaccgct
    10861 gaggagactc cggtcactgt cctcgccccg cctccccctt ccctcccctt ggggaccacc
    10921 gggcgccacg ccgcgaacgg taagtgccgc ggtcgtcggc gcctccgccc tccccctagg
    10981 gccccaattc ccagcgggcg cggcgcgcgg cccctccccc cgccgggcgc gcgcccgctg
    11041 ccccgccctt cgtggccgcc cggcgtgggc ggtgccaccc ctccccccgg cggccccgcg
    11101 cgcagctccc ggctccctcc cccttcggat gtggcttgag ctgtaggcgc ggaggggctc
    11161 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
    11221 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
    11281 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
    11341 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
    11401 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
    11461 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
    11521 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
    11581 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
    11641 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
    11701 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
    11761 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
    11821 gccaccgaga atcggacggg ggtagtctca agctggccgg cc
//
