LOCUS       sequence-120118-       14193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a firefly luciferase-enhanced green 
            fluorescent protein-cre recombinase fusion protein in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14193)
  AUTHORS   Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, 
            Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, 
            Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis
            PL, Jeffree RL, Johns TG, Boyd AW
  TITLE     A reference collection of patient-derived cell line and xenograft 
            models of proneural, classical and mesenchymal glioblastoma.
  JOURNAL   Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z.
  PUBMED    30894629
REFERENCE   2  (bases 1 to 14193)
  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: "Sci Rep."; 
            date: "2019-03-20"; volume: "9(1):4902. doi"; pages: " 
            10.1038/s41598-019-41277-z"
COMMENT     Sequence Label: pF CAG luc-EGFP-cre puro
FEATURES             Location/Qualifiers
     source          1..14193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..35
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        159..488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      339..474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        536..583
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     polyA_signal    1107..1228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1144..1163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1198..1217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1277..1293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1277..1293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1290..1312)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1301..1317
                     /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(1325..1331,1332..1349,1350..1355))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1370..1391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1508..1525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1679..2267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1759..1778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2438..3229,3230..3298))
                     /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     3061..3080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3299..3404)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3479..3498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3670..4049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4051..4249
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4204..4224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4267..4447
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4494..4619
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5112..5345
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5531..5575
                     /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    5873..5990
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6098..6401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6404..6679
                     /label=chicken beta-actin promoter
     intron          6680..7688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7612..7634
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7696..7715
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      7746..7755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7752..9401
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(7818..7836)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             join(9417..9419,9420..9422,9423..10133)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(9417..9419,9420..9422,9423..10133)
                     /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(9723..9742)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10070..10091
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             10173..11201
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     promoter        11254..11583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11434..11569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             11593..12192
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(11593..11612)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     12089..12109
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    12318..12906
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12371..12391)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12789..12800)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12909..12925)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12910..12926)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             13431..13611
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(13637..13654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13643..13867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(13913..14193,1..145)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13999..14018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
       61 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      121 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
      181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
      301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
      361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
      421 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
      481 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
      541 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
      601 catggcccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
      661 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
      721 gtggggtccg ggacgacggg accctgttca tcagcggcgg tccaggacca ggtggtgccg
      781 gacacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
      841 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
      901 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
      961 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     1021 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1081 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     1141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1201 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     1261 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1321 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1381 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1441 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1501 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1561 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1861 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2101 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2341 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2401 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2461 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2521 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2581 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2641 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2701 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2761 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2821 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2881 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2941 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3001 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3061 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3121 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3181 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3241 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3301 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3361 tatttgaatg tattttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3421 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     3481 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     3541 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     3601 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     3661 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     3721 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3781 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3841 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3901 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3961 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4021 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     4081 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4141 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4201 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
     4261 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     4321 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     4381 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4441 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     4501 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     4561 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     4621 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     4681 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     4741 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     4801 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     4861 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     4921 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     4981 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     5041 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     5101 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     5161 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     5221 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     5281 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     5341 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     5401 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     5461 gacagagaaa ttaacaatta cacaagctta atacactcct ttaattgaag aatcgcaaaa
     5521 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     5581 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     5641 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     5701 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     5761 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     5821 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     5881 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     5941 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     6001 acagggacag cagagatcca gtttggttaa ttaagactag ttattaatag taatcaatta
     6061 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggactat ttacggtaaa
     6241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6361 cttggcagta catctacgta ttagtcatcg ctattaccat gggtcgaggt gagccccacg
     6421 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     6481 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggcgcgcgc caggcggggc
     6541 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
     6601 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     6661 gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg ccccgtgccc cgctccgcgc
     6721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
     6781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc tcgtttcttt
     6841 tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc cctttgtgcg ggggggagcg
     6901 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc ccgcgctgcc
     6961 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgcg tgtgcgcgag
     7021 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct
     7081 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt
     7141 aacccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     7201 ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     7261 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc
     7321 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     7381 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg gcggagccga aatctgggag
     7441 gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt gcggcgccgg caggaaggaa
     7501 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct
     7561 cggggctgcc gcagggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     7621 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     7681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaaggat
     7741 ccgccgccac catggaagac gccaaaaaca taaagaaagg cccggcgcca ttctatcctc
     7801 tagaggatgg aaccgctgga gagcaactgc ataaggctat gaagagatac gccctggttc
     7861 ctggaacaat tgcttttaca gatgcacata tcgaggtgaa catcacgtac gcggaatact
     7921 tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat acaaatcaca
     7981 gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg ggcgcgttat
     8041 ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca
     8101 gtatgaacat ttcgcagcct accgtagtgt ttgtttccaa aaaggggttg caaaaaattt
     8161 tgaacgtgca aaaaaaatta ccaataatcc agaaaattat tatcatggat tctaaaacgg
     8221 attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct cccggtttta
     8281 atgaatacga ttttgtacca gagtcctttg atcgtgacaa aacaattgca ctgataatga
     8341 attcctctgg atctactggg ttacctaagg gtgtggccct tccgcataga actgcctgcg
     8401 tcagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg gatactgcga
     8461 ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc ggatatttga
     8521 tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt ttacgatccc
     8581 ttcaggatta caaaattcaa agtgcgttgc tagtaccaac cctattttca ttcttcgcca
     8641 aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct tctgggggcg
     8701 cacctctttc gaaagaagtc ggggaagcgg ttgcaaaacg cttccatctt ccagggatac
     8761 gacaaggata tgggctcact gagactacat cagctattct gattacaccc gagggggatg
     8821 ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt gtggatctgg
     8881 ataccgggaa aacgctgggc gttaatcaga gaggcgaatt atgtgtcaga ggacctatga
     8941 ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac aaggatggat
     9001 ggctacattc tggagacata gcttactggg acgaagacga acacttcttc atagttgacc
     9061 gcttgaagtc tttaattaaa tacaaaggat atcaggtggc ccccgctgaa ttggaatcga
     9121 tattgttaca acaccccaac atcttcgacg cgggcgtggc aggtcttccc gacgatgacg
     9181 ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag
     9241 agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg
     9301 tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga aaaatcagag
     9361 agatcctcat aaaggccaag aagggcggaa agtccaaatt ggctagcgga tccggaatgg
     9421 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     9481 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     9541 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     9601 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     9661 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     9721 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     9781 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     9841 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     9901 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     9961 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
    10021 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
    10081 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac
    10141 tcagatctcg agctcaagct tcgaattctg catccaattt actgaccgta caccaaaatt
    10201 tgcctgcatt accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt
    10261 tcagggatcg ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt
    10321 cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg
    10381 ttcgcgatta tcttctatat cttcaggcgc gcggtctggc agtaaaaact atccagcaac
    10441 atttgggcca gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca
    10501 atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg
    10561 caaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa
    10621 atagcgatcg ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc
    10681 tgttacgtat agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg
    10741 ggagaatgtt aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga
    10801 aggcacttag cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag
    10861 ctgatgatcc gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat
    10921 ctgccaccag ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat
    10981 tgatttacgg cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg
    11041 cccgtgtcgg agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc
    11101 aagctggtgg ctggaccaat gtaaatattg tcatgaacta tatccgtaac ctggatagtg
    11161 aaacaggggc aatggtgcgc ctgctggaag atggcgatta agctagcgtt aactctagag
    11221 atccgcggcc gcatcgattc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc
    11281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
    11341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    11401 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
    11461 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
    11521 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    11581 aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
    11641 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
    11701 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
    11761 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
    11821 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
    11881 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
    11941 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
    12001 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
    12061 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
    12121 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
    12181 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
    12241 catgcatcga ttagtccaat ttgttaaaga caggataacc tcgagggcgc gccgaattcg
    12301 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
    12361 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
    12421 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
    12481 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
    12541 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
    12601 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
    12661 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
    12721 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
    12781 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
    12841 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
    12901 ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc acaagtagca
    12961 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
    13021 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
    13081 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
    13141 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
    13201 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
    13261 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
    13321 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
    13381 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
    13441 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
    13501 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    13561 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
    13621 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
    13681 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
    13741 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
    13801 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
    13861 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
    13921 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    13981 gccagcgccc tagcgccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    14041 gctttcccgt cagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
    14101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    14161 tagacggttt ttcgcccttt gacgttggag tcc
//
