LOCUS       sequence-318915-       11435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GEF dead IQSEC1 mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11435)
  AUTHORS   Nacke M, Sandilands E, Nikolatou K, Roman-Fernandez A, Mason S, 
            Patel R, Lilla S, Yelland T, Galbraith LCA, Freckmann EC, McGarry L,
            Morton JP, Shanks E, Leung HY, Markert E, Ismail S, Zanivan S, Blyth
            K, Bryant DM
  TITLE     An ARF GTPase module promoting invasion and metastasis through 
            regulating phosphoinositide metabolism.
  JOURNAL   Nat Commun. 2021 Mar 12;12(1):1623. doi: 10.1038/s41467-021-21847-4.
  PUBMED    33712589
REFERENCE   2  (bases 1 to 11435)
  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: "Nat 
            Commun."; date: "2021-03-12"; volume: "12(1):1623. doi"; pages: " 
            10.1038/s41467-021-21847-4"
COMMENT     Sequence Label: pLEX304-GFP-IQSEC1 v2-E620K GEF dead mutant
FEATURES             Location/Qualifiers
     source          1..11435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(638..640,641..643,644..1354)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(638..640,641..643,644..1354)
                     /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(683..704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(944..963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1291..1312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4463..5051
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4516..4536)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4934..4945)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5123..5356
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5434..5555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5471..5490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5525..5544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5595..5730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5750..5769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5751..5769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5779..5796)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5779..5795)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5788..5810)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5937..6392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6024..6043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6234..6255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6418..6522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6523..6591,6592..7383)
                     /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(6741..6760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7554..8142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8043..8062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8296..8313
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8430..8451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8466..8471,8472..8489,8490..8496)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8504..8520
                     /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)."
     primer_bind     8509..8531
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8528..8544
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8528..8544
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8563..8583
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8565..8583
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8611..8837
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8838..9018
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9065..9190
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9683..9916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10101..10145
                     /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        10320..10820
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     10739..10757
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             10832..11227
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11287..11404
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tttggcgccg gctcgagggg gcccggggtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
      661 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      721 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      781 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
      841 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      901 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      961 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1021 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1081 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1141 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1201 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1261 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1321 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc
     1381 ttacatgtgg tgcctgcact gcaactcgga gaggacccag tcccttctgg agctagagct
     1441 tgacagcggc gtcgagggcg aggcccccag cagtgagact ggcacatccc tggacagccc
     1501 ctcagcctac ccccagggcc ccttggtgcc cggttccagc ctgagcccgg atcactacga
     1561 gcacacgtca gtgggagcct atgggctgta ctcggggccg ccggggcaac agcagcgcac
     1621 gcggaggccc aagctgcagc actcgacctc catcctgcgc aagcaggctg aggaggaggc
     1681 catcaagcgc tcacgctcac tctccgagag ctatgagctc tcctcggacc tgcaggacaa
     1741 gcaggtggag atgctagagc gaaagtatgg ggggcgcctg gtaacccgcc atgcggcccg
     1801 caccatccag acggcgtttc gccaatatca aatgaataaa aacttcgagc gcttgcgcag
     1861 ctccatgtca gagaaccgca tgtcacgccg gattgtgctg tccaacatga ggatgcagtt
     1921 ctcctttgag gggcctgaga aagtgcacag ctcctacttc gaggggaagc aggtctcagt
     1981 gactaacgac ggctcccagc tgggagccct ggtgtcccct gagtgtggtg acctcagcga
     2041 gcccaccacc ctcaagtctc cggccccctc cagtgacttt gcggacgcca tcaccgagct
     2101 ggaggacgcc ttctctaggc aagtgaaatc actggccgag tccatcgacg atgccctcaa
     2161 ctgccgcagc ctgcacactg aggaggcacc ggccctggat gcggcgcggg cccgggacac
     2221 cgaaccccag acagccctgc acggcatgga ccaccgcaaa ctggacgaga tgacggcctc
     2281 gtacagtgat gtcaccctgt acatcgatga ggaggagctg tcgccccctc tgcccctctc
     2341 gcaggcaggg gaccggccgt ccagcaccga gtcggacctg cggctacggg ctgggggcgc
     2401 agccccagac tactgggccc tggcccacaa agaggacaag gctgacacgg acacgagctg
     2461 ccggagcacg ccgtcgctgg agcggcagga gcagcggctg cgggtggagc atctgccgct
     2521 gctcaccatc gagccaccca gcgacagctc tgtggacctt agtgaccgct cggagcgggg
     2581 gtcactcaag aggcagagtg cttacgagcg cagccttggc gggcagcagg gcagtcccaa
     2641 gcatggtccc cacagcggcg cccccaagag cctcccccgg gaggagcctg agttgcggcc
     2701 ccggcccccc aggcccctgg acagccactt ggccatcaat ggctcagcca accggcagag
     2761 caagtctgag tcggactact cagacggtga caatgacagc atcaacagca cgtccaactc
     2821 caacgatacc atcaactgca gctccgagtc atcgtcccgt gacagcctgc gggagcagac
     2881 gctcagcaag cagacctacc acaaggaggc ccgcaacagc tgggactcgc ctgcctttag
     2941 caacgatgtc atccgcaaga ggcactaccg catcggcctg aacctcttca acaagaagcc
     3001 tgagaaggga gtccagtacc tcatcgagcg tggctttgtg cccgacacgc ccgtcggggt
     3061 ggcccacttc ctgctgcagc gcaagggcct cagccggcag atgatcggcg agttcctggg
     3121 caaccggcag aagcagttca accgtgacgt gctcgactgc gtcgtggacg agatggactt
     3181 ctctaccatg gagctggatg aggccctcag gaaattccag gcgcacatcc gtgtccaagg
     3241 gaaggctcag aaagtggagc ggctcataga ggcgttcagc cagcgctact gcatctgcaa
     3301 ccctggggtg gtgcggcaat tccggaaccc agacaccatt ttcatcctgg ccttcgccat
     3361 catcctgctg aacaccgaca tgtacagccc caatgtcaag cccgagcgga aaatgaagct
     3421 agaggacttc atcaagaacc tccgaggtgt ggacgatggt gaggacattc cccgtgagat
     3481 gctgatgggg atctatgaac ggatccgtaa gcgagagcta aagaccaatg aggaccatgt
     3541 gtcccaggtg cagaaggtgg agaagctcat tgtggggaaa aagccgatcg gatccctgca
     3601 tcccgggctc ggctgtgtgc tctctctgcc ccaccgtcgg ttggtctgct actgccggct
     3661 ctttgaggtt ccagacccaa acaagcccca gaaactcgga ctacaccagc gagaaatctt
     3721 cctgttcaac gacctcctgg tggtcaccaa gatcttccag aagaagaaga actcggtgac
     3781 gtacagcttc cgacagtcct tctccttgta cggcatgcag gtcctgctct tcgagaacca
     3841 gtactacccc aatggcatcc ggctcacctc gtctgtcccc ggagcagata tcaaagtgtt
     3901 aataaacttc aacgccccca accctcaaga ccggaagaaa ttcaccgatg acctgcggga
     3961 gtccattgcg gaagtccaag agatggagaa gcacaggata gagtcggagc tcgagaagca
     4021 gaaaggcgtc gtgcggccca gcatgtccca gtgctctagc ctcaaaaagg agtcgggcaa
     4081 cggaacactg agccgggcct gcctggacga cagctatgcc agcggtgagg gcctcaagcg
     4141 cagcgccctc agcagctccc tgcgggacct ctcggaagcc gggaagcgag ggcgtcgcag
     4201 cagtgcggga tcgctagaga gcaatgtgga atttcagcct ttcgagccac tgcagccgtc
     4261 agtgctgtgc tcctaagaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     4321 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4381 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttgcta
     4441 accggtggcg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4501 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4561 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4621 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4681 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4741 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4801 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4861 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4921 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4981 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5041 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5101 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     5161 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5221 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     5281 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     5341 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
     5401 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     5461 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5521 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     5581 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5641 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5701 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     5761 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5821 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5881 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     5941 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6001 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     6061 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     6121 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6181 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6241 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6301 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6361 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     6421 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6541 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6601 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6661 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6721 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6781 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6841 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6901 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6961 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7021 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7081 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7141 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7201 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7261 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7321 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7381 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7441 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7501 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7561 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7621 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7681 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7741 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7801 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7861 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7921 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7981 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8041 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8101 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8161 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8221 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8281 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     8341 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8401 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8461 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8521 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     8581 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     8641 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     8701 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     8761 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     8821 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8881 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8941 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9001 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     9061 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     9121 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     9181 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     9241 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     9301 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     9361 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     9421 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     9481 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     9541 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     9601 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     9661 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     9721 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     9781 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     9841 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     9901 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     9961 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
    10021 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
    10081 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
    10141 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
    10201 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
    10261 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccct
    10321 tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt
    10381 ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg
    10441 tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc
    10501 taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac
    10561 gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc
    10621 gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag
    10681 gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt
    10741 gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa
    10801 tcaccgacct ctctccccag ggggtaccac catggccaag cctttgtctc aagaagaatc
    10861 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
    10921 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
    10981 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
    11041 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
    11101 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
    11161 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
    11221 ggagggcctg cagctgcagt agtaagaatt ctagatcttg agacaaatgg cagtattcat
    11281 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
    11341 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
    11401 ttttcgggtt tattacaggg acagcagaga tccac
//
