LOCUS       sequence-377366-       10060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Tiam1(DH-PH) fused to miRFP703-eDHFR(69K6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10060)
  AUTHORS   Hino N, Matsuda K, Jikko Y, Maryu G, Sakai K, Imamura R, Tsukiji S, 
            Aoki K, Terai K, Hirashima T, Trepat X, Matsuda M
  TITLE     A feedback loop between lamellipodial extension and HGF-ERK 
            signaling specifies leader cells during collective cell migration.
  JOURNAL   Dev Cell. 2022 Oct 10;57(19):2290-2304.e7. doi: 
            10.1016/j.devcel.2022.09.003. Epub 2022 Sep 28.
  PUBMED    36174555
REFERENCE   2  (bases 1 to 10060)
  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"; doi: 
            "10.1016/j.devcel.2022.09.003"; journalName: "Dev Cell"; date: 
            "2022-10-10- 10"; volume: "57"; issue: "19"; pages: "2290-2304.e7"
COMMENT     Sequence Label: pPBbleo-miRFP703-eDHFR(69K6)-Tiam1(DH-PH)
FEATURES             Location/Qualifiers
     source          1..10060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1668..2684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2608..2630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2692..2711
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2751..2760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2757..3701
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     misc_feature    6178..6727
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6322..6339)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6549..6568
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6730..7104
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(7144..7161)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7150..7374
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7421..7437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7445..7461
                     /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(7469..7475,7476..7493,7494..7499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7514..7535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7769..7831
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7872..7892)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7872..7890)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7911..7927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7935..7951
                     /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(7959..7965,7966..7983,7984..7989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8004..8025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8142..8159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8313..8901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8393..8412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9072..9863,9864..9932))
                     /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     9695..9714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9933..10037)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     1441 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     1501 gggggggggg gsgcgcgccr ggsggggsgg gggggggsgg gggggggggs ggggsgrggs
     1561 ggrgaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1621 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1681 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1741 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1801 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1861 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1921 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1981 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     2041 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     2101 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     2161 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     2221 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     2281 ccggggrggg ctcgggggrg gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     2341 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     2401 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     2461 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     2521 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2581 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2641 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     2701 tggttattgt gctgtctcat cattttggca aagaattggg atccgaattc gccaccatgg
     2761 tagcaggtca tgcctctggc agccccgcat tcgggaccgc ctctcattcg aattgcgaac
     2821 atgaagagat ccacctcgcc ggctcgatcc agccgcatgg cgcgcttctg gtcgtcagcg
     2881 aacatgatca tcgcgtcatc caggccagcg ccaacgccgc ggaatttctg aatctcggaa
     2941 gcgtactcgg cgttccgctc gccgagatcg acggcgatct gttgatcaag atcctgccgc
     3001 atctcgatcc caccgccgaa ggcatgccgg tcgcggtgcg ctgccggatc ggcaatccct
     3061 ctacggagta ctgcggtctg atgcatcggc ctccggaagg cgggctgatc atcgaactcg
     3121 aacgtgccgg cccgtcgatc gatctgtcag gcacgctggc gccggcgctg gagcggatcc
     3181 gcacggcggg ttcactgcgc gcgctgtgcg atgacaccgt gctgctgttt cagcagtgca
     3241 ccggctacga ccgggtgatg gtgtatcgtt tcgatgagca aggccacggc ctggtattct
     3301 ccgagtgcca tgtgcctggg ctcgaatcct atttcggcaa ccgctatccg tcgtcgctgg
     3361 tcccgcagat ggcgcggcag ctgtacgtgc ggcagcgcgt ccgcgtgctg gtcgacgtca
     3421 cctatcagcc ggtgccgctg gagccgcggc tgtcgccgct gaccgggcgc gatctcgaca
     3481 tgtcgggctg cttcctgcgc tcgatgtcgc cgatccatct gcagttcctg aaggacatgg
     3541 gcgtgcgcgc caccctggcg gtgtcgctgg tggtcggcgg caagctgtgg ggcctggttg
     3601 tctgtcacca ttatctgccg cgcttcatcc gtttcgagct gcgggcgatc tgcaaacggc
     3661 tcgccgaaag gatcgcgacg cggatcaccg cgcttgagag cctgtacaag tctgggtccg
     3721 cagccagtag cgccggaagt atgatcagtc tgattgcggc gttagcggta gatcgcgtta
     3781 tcggcatgga aaacgccatg ccgtggaacc tgcctgccga tctcgcctgg tttaaacgca
     3841 acaccttaaa taaacccgtg attatgggcc gccatacctg ggaatcaatc ggtcgtccgt
     3901 tgccaggacg caaaaatatt atcctcagca gtcaaccggg tacggacaaa aaaaagaaaa
     3961 agaaagatcg cgtaacgtgg gtgaagtcgg tggatgaagc catcgcggcg tgtggtgacg
     4021 taccagaaat catggtgatt ggcggcggtc gcgtttatga acagttcttg ccaaaagcgc
     4081 aaaaactgta tctgacgcat atcgacgcag aagtggaagg cgacacccat ttcccggatt
     4141 acgagccgga tgactgggaa tcggtattca gcgaattcca cgatgctgat gcgcagaact
     4201 ctcacagcta ttgctttgag attctggagc ggcggctgta caagtccgga ctcagatctc
     4261 gacaaggtag tggtgctggc tctggtgctg gtagtggcgc tggttccggt gctggctctg
     4321 gcgcgcctcg agcaatgaac ccctctgacc agagcccatc tcctcaggac tccacggggc
     4381 ctcagctggc gaccatgaga caactctcgg atgcagataa gctgcgcaag gtgatctgcg
     4441 agctcctgga gacggagcgc acctacgtga aggatttaaa ctgtcttatg gagagatacc
     4501 taaagcctct tcaaaaagaa acttttctca cccaggatga gcttgacgtg ctttttggaa
     4561 atttaacgga aatggtagag tttcaagtag aattccttaa aactctagaa gatggagtga
     4621 gactggtacc tgatttggaa aagcttgaga aggttgatca atttaagaaa gtgctgttct
     4681 ctctgggggg atcattcctg tattatgctg accgcttcaa gctctacagt gccttctgcg
     4741 ccagccacac aaaagttccc aaggtcctgg tgaaagccaa gacagacacg gctttcaagg
     4801 cattcttgga tgcccagaac ccgaagcagc agcactcatc cacgctggag tcgtacctca
     4861 tcaagcccat ccagaggatc ctcaagtacc cacttctgct cagggagctg ttcgccctga
     4921 ccgatgcgga gagcgaggag cactaccacc tggacgtggc catcaagacc atgaacaagg
     4981 ttgccagtca catcaatgag atgcagaaaa tccatgaaga gtttggggct gtgtttgacc
     5041 agctgattgc tgaacagact ggtgggaaaa aagaggttgc agatctgagc atgggagacc
     5101 tgcttttgca cactaccgtg atctggctga acccgccggc ctcgctgggc aagtggaaaa
     5161 aggaaccaga gttggcagca ttcgtcttca aaactgctgt ggtccttgtg tataaagatg
     5221 gttccaaaca gaagaagaaa cttgtaggat ctcacaggct ttccatttat gaggactggg
     5281 accccttcag atttcgacac atgatcccca cggaagcgct gcaggttcga gctttggcga
     5341 gtgcagatgc agaggcaaat gccgtgtgtg aaattgtcca tgtaaaatcc gagtctgaag
     5401 ggaggccgga gagggtcttt cacttgtgct gcagctcccc agagagccga aaggatttcc
     5461 taaaggctgt gcattcaatc ctgcgtgata agcacagaag acagctcctc aaaaccgaga
     5521 gccttccctc atcccagcaa tatgtccctt ttggaggcaa aagattgtgt gcactgaagg
     5581 gggccaggcc ggccatgagc agggcagtgt ctgccccaag caagtctctt gggaggagga
     5641 ggcggcggct ggctcgaaac aggtttacca ttgattctga tgccgtctcc gcaagcagcc
     5701 cggagaaaga gtcccagcag ccccccggtg gcggggacac tgaccgatgg gtagaggagc
     5761 agtttgatct tgctcagtat gaggagcaag atgacatcaa ggagacagac atcctcagtg
     5821 acgatgatga gttctgtgag tccgtgaagg gtgcctcagt ggacagagac ctgcaggagc
     5881 ggcttcaggc cacctccatc agtcagcggg aaagaggccg gaaaaccctg gatagtcacg
     5941 cgtcccgcat ggcacagctc aagaagcaag ctgccctgtc ggggatcaat ggaggcctgg
     6001 agagcgcaag cgaggaagtc atttgggtta ggcgtgaaga ctttgccccc tccaggaaac
     6061 tgaacactga gatctgagcg gccgctctag agtcgacggg ccgcggtaac aattgttaac
     6121 taacttaagc tagcaacggt ttccctctag cgggatcaat tccgcccccc ccccctaacg
     6181 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     6241 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     6301 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     6361 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     6421 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     6481 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     6541 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     6601 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     6661 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     6721 gataatacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     6781 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     6841 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     6901 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     6961 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     7021 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     7081 ttcgtggccg aggagcagga ctgaaagtcg agatgcatcg atgatctaga gctcgctgat
     7141 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7201 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7261 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7321 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     7381 aggcggaaag aacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7441 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7501 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7561 tccagtcggg aaacctgtcg tgccagcgga tcttaaaagt tttgttactt tatagaagaa
     7621 attttgagtt tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat
     7681 ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat
     7741 aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa
     7801 cgtacgtcac aatatgatta tctttctagg gttaatctag tatacgcgtt taattaatcc
     7861 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     7921 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7981 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     8041 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     8101 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     8161 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     8221 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     8281 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8341 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8401 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8461 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8521 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8581 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8641 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8701 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8761 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8821 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8881 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8941 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     9001 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     9061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     9121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     9181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     9241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9361 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     9421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9661 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    10021 caaatagggg ttccgcgcac atttccccga aaagtgccac
//
