LOCUS       sequence-416585-       12298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To monitor the status of TGF-alpha, the plasmid encodes a 
            recombinant TGF-alpha fused extracellularly to the mScarlet 
            fluorescent protein and intracellularly to the mNeonGreen 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12298)
  AUTHORS   Deguchi E, Lin S, Hirayama D, Matsuda K, Tanave A, Sumiyama K, 
            Tsukiji S, Otani T, Furuse M, Sorkin A, Matsuda M, Terai K
  TITLE     Low-affinity ligands of the epidermal growth factor receptor are 
            long-range signal transmitters in collective cell migration of 
            epithelial cells.
  JOURNAL   Cell Rep. 2024 Nov 14;43(11):114986. doi: 
            10.1016/j.celrep.2024.114986.
  PUBMED    39546398
REFERENCE   2  (bases 1 to 12298)
  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: "Cell Rep.";
            date: "2024-11-14"; volume: "43(11):114986. doi"; pages: " 
            10.1016/j.celrep.2024.114986"
COMMENT     Sequence Label: pCSIIpuro-TGFa-ScNeo
FEATURES             Location/Qualifiers
     source          1..12298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1676..1909
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2094..2138
                     /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    2412..2529
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2585..3747
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2816..3738
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3695..3715
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3904..3930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3931..4626
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(4064..4082)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4332..4351
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5807..6356
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5951..5968)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6178..6197
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6359..6958
                     /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(6359..6378)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6855..6875
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6993..7581
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7046..7066)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7464..7475)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8100..8280
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8306..8323)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8312..8536
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8582..9010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8669..8688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8879..8900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9019..9039)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9024..9353
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9204..9339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9266..9285
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9401..9448
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9467..9841
                     /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"
     polyA_signal    9971..10092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10008..10027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10062..10081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10141..10157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10141..10157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10154..10176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10165..10181
                     /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(10189..10195,10196..10213,10214..10219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10234..10255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10372..10389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10543..11131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10623..10642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11302..12093,12094..12162))
                     /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     11925..11944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12163..12267)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gggccgctga tcttcagacc tggaggagga gatatgaggg
     1561 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1621 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1681 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1741 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1801 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1861 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1921 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1981 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     2041 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     2101 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2161 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2221 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2281 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2341 gtggagagag agacagagac agatccattc gattagtgaa cggatctaca aatggcagta
     2401 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2461 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2521 caaaattttc gggtttatta cagggacagc agaaattcac tttgaattaa ttcaagcttc
     2581 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2641 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     2701 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2761 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     2821 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2881 aattacttcc acctggctcc agtacgtgat tcttgatccc gagctggagc caggggcggg
     2941 ccttgcgctt taggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     3001 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     3061 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     3121 gtaaatgcgg gccaggatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     3181 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     3241 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     3301 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     3361 gaaagatggc cgcttcccgg ccctgctcca gggggctcaa aatggaggac gcggcgctcg
     3421 ggagagcggg cgggtgagtc acccacacaa aggaaagggg cctttccgtc ctcagccgtc
     3481 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctggagc
     3541 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     3601 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     3661 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     3721 ttttttcttc catttcaggt gtcgtgaaca cgctaccggt ccgaattctc caccatggtc
     3781 ccctcggctg gacagctcgc cctgttcgct ctgggtattg tgttggctgc gtgccaggcc
     3841 ttggagaaca gcacgtcccc gctgagtgca gacccgcccg tggctgcagc agtggtgtcc
     3901 cattacccat acgatgttcc ggattacgct atggtgagca agggcgaggc agtgatcaag
     3961 gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc
     4021 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     4081 aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc
     4141 agggccttca ccaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag
     4201 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag
     4261 gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc
     4321 cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg
     4381 ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac
     4441 ggaggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg
     4501 cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc
     4561 gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg
     4621 tacaagggtg gtggtagtgg tggtggttca gtgtcccatt ttaatgactg cccagattcc
     4681 cacactcagt tctgcttcca tggaacctgc aggtttttgg tgcaggagga caagccagca
     4741 tgtgtctgcc attctgggta cgttggtgca cgctgtgagc atgcggacct cctggccgtg
     4801 gtggctgcca gccagaagaa gcaggccatc accgccttgg tggtggtctc catcgtggcc
     4861 ctggctgtcc ttatcatcac atgtgtgctg atacactgct gccaggtccg aaaacactgt
     4921 gagtggtgcc gggccctcat ctgccggcac gagaagccca gcgccctcct gaagggaaga
     4981 accgcttgct gccactcaga aacaatggta agcaaggggg aggaagataa tatggcttcc
     5041 cttccagcaa cgcacgaatt gcatatcttt ggaagtatta atggagttga tttcgacatg
     5101 gtggggcagg gtacgggaaa cccaaacgat ggctatgaag aacttaacct caagtctacc
     5161 aaaggcgatc tccaattctc accctggata ttggtcccac acatcggcta cgggttccat
     5221 caatacctgc cttatcctga cgggatgagt ccttttcaag ctgctatggt tgacggaagt
     5281 ggctaccaag ttcaccgcac aatgcaattt gaggatggtg catctcttac ggttaattac
     5341 cgatatacat atgagggttc acacatcaag ggggaagctc aggtcaaggg cactggattt
     5401 cccgcagatg gcccagttat gaccaattct ctgaccgcag ccgattggtg tcgaagcaag
     5461 aagacctacc ctaacgacaa gactatcatc tcaaccttca aatggtccta caccacaggt
     5521 aacggaaagc ggtataggtc tacagctcgg acaacatata cctttgcgaa gcctatggcc
     5581 gctaactatt tgaaaaacca accgatgtat gtttttcgga agaccgaatt gaaacattca
     5641 aaaacggagt tgaatttcaa agagtggcag aaggcattca ccgacgtaat gggtatggat
     5701 gagttgtata aagtggtcta agcggccgcg gctctagagg atccgttaac taacttaagc
     5761 tagcaacggt ttccctctag cgggatcaat tccgcccccc cccctaacgt tactggccga
     5821 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     5881 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     5941 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     6001 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     6061 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     6121 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     6181 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     6241 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     6301 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg ataataccat
     6361 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     6421 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     6481 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     6541 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     6601 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     6661 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6721 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6781 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6841 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6901 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgact
     6961 agagttaaca tcgagggatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     7021 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     7081 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     7141 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     7201 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     7261 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     7321 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     7381 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     7441 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     7501 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     7561 ccctttgggc cgcctccccg catcgatacc gtcgagacct agaaaaacat ggagcaatca
     7621 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     7681 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     7741 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     7801 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     7861 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca
     7921 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct
     7981 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga
     8041 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg
     8101 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     8161 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     8221 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     8281 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     8341 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     8401 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     8461 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     8521 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     8581 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     8641 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     8701 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     8761 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     8821 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     8881 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     8941 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     9001 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     9061 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     9121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     9181 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     9241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     9301 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     9361 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     9421 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     9481 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     9541 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     9601 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     9661 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     9721 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     9781 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     9841 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     9901 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     9961 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
    10021 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
    10081 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
    10141 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    10201 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    10261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
    10321 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
    10381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
    10441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    10501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    10561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    10621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    10681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    10741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    10801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    10861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    10921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    10981 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    11041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    11101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    11161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
    11221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    11281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    11341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    11401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
    11461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
    11521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
    11581 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
    11641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
    11701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
    11761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
    11821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
    11881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    11941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    12001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    12061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    12121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    12181 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    12241 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
