LOCUS       sequence-421035-        8686 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 8686)
  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 8686)
  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: pPBbleo-TGFa-ScNeo
FEATURES             Location/Qualifiers
     source          1..8686
                     /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"
     CDS             2886..2912
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2913..3608
                     /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(3046..3064)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3314..3333
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4804..5353
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4948..4965)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5175..5194
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5356..5730
                     /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(5770..5787)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5776..6000
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6047..6063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6071..6087
                     /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(6095..6101,6102..6119,6120..6125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6140..6161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6395..6457
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6498..6518)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6498..6516)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6537..6553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6561..6577
                     /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(6585..6591,6592..6609,6610..6615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6630..6651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6768..6785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6939..7527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7019..7038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7698..8489,8490..8558))
                     /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     8321..8340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8559..8663)
                     /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 gcgcgcgccr ggsggggsgg ggsggggsgr ggggsggggs ggggsgrggc
     1561 ggagaggtgc 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 ctcgggggag 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 tccaccatgg
     2761 tcccctcggc tggacagctc gccctgttcg ctctgggtat tgtgttggct gcgtgccagg
     2821 ccttggagaa cagcacgtcc ccgctgagtg cagacccgcc cgtggctgca gcagtggtgt
     2881 cccattaccc atacgatgtt ccggattacg ctatggtgag caagggcgag gcagtgatca
     2941 aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga
     3001 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     3061 ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct
     3121 ccagggcctt caccaagcac cccgccgaca tccccgacta ctataagcag tccttccccg
     3181 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc
     3241 aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact
     3301 tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc
     3361 ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg
     3421 acggaggccg ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga
     3481 tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca
     3541 ccgtggtgga acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc
     3601 tgtacaaggg tggtggtagt ggtggtggtt cagtgtccca ttttaatgac tgcccagatt
     3661 cccacactca gttctgcttc catggaacct gcaggttttt ggtgcaggag gacaagccag
     3721 catgtgtctg ccattctggg tacgttggtg cacgctgtga gcatgcggac ctcctggccg
     3781 tggtggctgc cagccagaag aagcaggcca tcaccgcctt ggtggtggtc tccatcgtgg
     3841 ccctggctgt ccttatcatc acatgtgtgc tgatacactg ctgccaggtc cgaaaacact
     3901 gtgagtggtg ccgggccctc atctgccggc acgagaagcc cagcgccctc ctgaagggaa
     3961 gaaccgcttg ctgccactca gaaacaatgg taagcaaggg ggaggaagat aatatggctt
     4021 cccttccagc aacgcacgaa ttgcatatct ttggaagtat taatggagtt gatttcgaca
     4081 tggtggggca gggtacggga aacccaaacg atggctatga agaacttaac ctcaagtcta
     4141 ccaaaggcga tctccaattc tcaccctgga tattggtccc acacatcggc tacgggttcc
     4201 atcaatacct gccttatcct gacgggatga gtccttttca agctgctatg gttgacggaa
     4261 gtggctacca agttcaccgc acaatgcaat ttgaggatgg tgcatctctt acggttaatt
     4321 accgatatac atatgagggt tcacacatca agggggaagc tcaggtcaag ggcactggat
     4381 ttcccgcaga tggcccagtt atgaccaatt ctctgaccgc agccgattgg tgtcgaagca
     4441 agaagaccta ccctaacgac aagactatca tctcaacctt caaatggtcc tacaccacag
     4501 gtaacggaaa gcggtatagg tctacagctc ggacaacata tacctttgcg aagcctatgg
     4561 ccgctaacta tttgaaaaac caaccgatgt atgtttttcg gaagaccgaa ttgaaacatt
     4621 caaaaacgga gttgaatttc aaagagtggc agaaggcatt caccgacgta atgggtatgg
     4681 atgagttgta taaagtggtc taagcggccg ctctagagtc gacgggccgc ggtaacaatt
     4741 gttaactaac ttaagctagc aacggtttcc ctctagcggg atcaattccg cccccccccc
     4801 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     4861 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     4921 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     4981 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     5041 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     5101 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     5161 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     5221 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     5281 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     5341 aaacacgata ataccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     5401 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     5461 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     5521 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     5581 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     5641 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     5701 gtgcacttcg tggccgagga gcaggactga aagtcgagat gcatcgatga tctagagctc
     5761 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5821 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5881 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5941 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     6001 cttctgaggc ggaaagaacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     6061 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6121 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6181 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatctt aaaagttttg ttactttata
     6241 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
     6301 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
     6361 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
     6421 ctttaacgta cgtcacaata tgattatctt tctagggtta atctagtata cgcgtttaat
     6481 taatccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     6541 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6601 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6661 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6721 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6781 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7681 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7741 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7801 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7861 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7921 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7981 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8041 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8101 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8161 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8221 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8281 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8341 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8401 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8461 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8521 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
