LOCUS       sequence-460021-        8550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of genetically encoded calcium indicator in zebrafish 
            hepatocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8550)
  AUTHORS   Pozo-Morales M, Garteizgogeascoa I, Perazzolo C, So J, Shin D, Singh
            SP
  TITLE     In vivo imaging of calcium dynamics in zebrafish hepatocytes.
  JOURNAL   Hepatology. 2023 Mar 1;77(3):789-801. doi: 10.1002/hep.32663. Epub 
            2023 Feb 17.
  PUBMED    35829917
REFERENCE   2  (bases 1 to 8550)
  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: "10.1002/hep.32663";
            journalName: "Hepatology"; date: "2023-03-1- 1"; volume: "77"; 
            issue: "3"; pages: "789-801"
COMMENT     Sequence Label: fabp10a:GCaMP6s; cryaa:mCherry
FEATURES             Location/Qualifiers
     source          1..8550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..62
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        43..61
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2293..2302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2299..2307,2308..2325,2326..2328,2329..2361,2362..2364,
                     2365..2388,2389..2415,2416..2472,2473..2478,2479..2748,
                     2749..2766,2767..3201,3202..3207,3208..3651)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      2395..2404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2685..2706
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2812..2833)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3073..3092)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3949..4083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4040..4059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4101..4148
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    4172..4379
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4368..4385
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4403..5113)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4681..4700)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4950..4968
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(5679..5712)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5731..5751)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5731..5749)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5782..5799)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5782..5798)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5791..5813)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5940..6368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6027..6046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6237..6258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6421..6525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6526..6594,6595..7386)
                     /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(6744..6763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7557..8145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8046..8065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8299..8316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8433..8454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8469..8474,8475..8492,8493..8499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8507..8523
                     /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     8512..8534
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8531..8547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8531..8547
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattacgcca agcgcgttag ggataacagg gtaatgcgcg cgtaatacga ctcactatag
       61 ggcgaattgg agctctgcag aagagcatct ctgaacacac aacacgtcca accttgaggc
      121 agatgggcta cagcagcaga agaccacacc gggtgccgct cctgtcagct aagaacagga
      181 aactgaggct acaattcaca cagactcacc aaaactggac gatatattat tagcccccct
      241 tagaattttt catttgataa tatttttctt ctggcgaaag cctcatttgt tttatatatt
      301 atagaataaa attagttttt aatagttttt atgccatttt aaggtcaata ttattagccc
      361 ctttaagcta ttttttttcg atagtctaca gaacaaacca tcggtataca atgacttgcc
      421 taattaccct aacctgccta gttaccctaa ttaacctagt taagccttta aatgtcactt
      481 taagctgtat agaagtgtct tgaagaatat ctagtctaat attattgact gtcatcatgg
      541 caaagataaa ataaatcagt tattaaaact attatgatta gaaatgtgct gaaacaatct
      601 gctctccgat aaacagaaat tgaacaaaat aaacaggggg gctaataaat ttaaggggtt
      661 aaataattct gattgcaaaa aaaatgatgt ctgcaaaact gttgcaaata atttatttgt
      721 gttgatttta agcaaacaaa ttaaatttaa taaaatacaa cttaatctgt ttgtttaaat
      781 tcagccctaa taaattgttt acagccactt aacgtaaaaa aattgagtaa atccaaggaa
      841 tcatctctga ataatttttt tcagtgtata tatatatata tatattctta caaaacaact
      901 catttacttt agtttaattt tcaggggcaa aaaactaaag taatcgacgt tgcttgaata
      961 aaaagtgtaa ttaagggaat gaggtaacat ttaaccatgt gtcaatgcag tttaaatatg
     1021 ccagttagtg gtatatgttt aaatggtaag ctattcaaaa ctttaaacta acttaaccag
     1081 ccttttgttg tcagactgaa cagactttcc atctgcatta ttagagacta atctttggct
     1141 ggatgaatga ttcatctgct gatatttcag aatagacaga ttgaggctgt ttctaatatg
     1201 attatgcaac ctgagggtga ttatttgaag caaactccac agaccagcag gtcattgacc
     1261 gtcgtgtgtt caaacagagc agaaacattt gcaaaactgg tctgacagga gaatccagtc
     1321 cagcacaaca catatgctga gcaaactgaa tcaatcctgc aggtcaactc tcgtgcttta
     1381 agtttattaa gattatttta tttatttatt attttattta tctatttatt tatttagttg
     1441 tttatttagc aattatttgt ttatttattt gtttatttat tcctgcagat catgccttgt
     1501 gcctttttac atttaattta atttttaatt taatttcctt ttactttttt ttattttttt
     1561 attttatttt attttacagt ctgacatata ctsmamtaaa accctcagat catgtcttat
     1621 gcatttmatt tnattttttn ttttttttnt ntttatttta ttwtttattt tattttattw
     1681 tttattttat tttattttat tttattttac agtctgacat actggaccaa taaatctctc
     1741 agatcatgtc tcatgcattt tacttttatt ttattagaat tagaaagatc aaaggaacaa
     1801 cttttaaaat attaattctg tatcaaaatc tcttttgata catttaattg atttaaaaaa
     1861 gcagttcacc caagaaacat ttcctcacag tcgaatggtt gtaaactttt atgaattact
     1921 ttcacagaaa aagatttttg gaagaatatt ggaaaaaaag cagccattga cttccatagt
     1981 aacaacaaaa aatactatgg aagtcaatgg ctgttttttc accattcggt atcttcattc
     2041 tggagcagaa ttttttgggt gatctgtccc tttaagtcgt caaatcctgg tgcaatattc
     2101 cacatgcact gacgagtctt tatttttggt ctgctactgc tgtgtgtgtg agggcattta
     2161 cctcatcttc tgctggagtt gatgaacggt gggttgttca aacagcagca ggtcattgac
     2221 tgaactcctc tcgatataaa agctgcagat ctgaagctga ccttcacttt gtgttgatcg
     2281 ctagtggatc tcgccaccat gggttctcat catcatcatc atcatggtat ggctagcatg
     2341 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgccacc
     2401 atggtcgact catcacgtcg taagtggaat aagacaggtc acgcagtcag agctataggt
     2461 cggctgagct cactcgagaa cgtctatatc aaggccgaca agcagaagaa cggcatcaag
     2521 gcgaacttcc acatccgcca caacatcgag gacggcggcg tgcagctcgc ctaccactac
     2581 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2641 gtgcagtcca aactttcgaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2701 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     2761 gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2821 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc
     2881 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2941 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3001 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacatcca ggagcgcacc
     3061 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3121 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3181 gggcacaagc tggagtacaa cctgccggac caactgactg aagagcagat cgcagaattt
     3241 aaagaggctt tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg
     3301 gggacggtga tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc
     3361 aatgaagtag atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg
     3421 gcaagaaaaa tgaaatacag ggacacggaa gaagaaatta gagaagcgtt cggtgtgttt
     3481 gataaggatg gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt
     3541 ggagagaagt taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg
     3601 gatggtcagg taaactacga agagtttgta caaatgatga cagcgaagtg agcggccgcc
     3661 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3721 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3781 cgcggggatc tcatgctgga gttcttcgcc caccccccgg atctaagctc tagataagta
     3841 atgatcataa tcagccatat cacatctgta gaggttttac ttgctttaaa aaacctccca
     3901 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3961 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4021 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4081 atccgggggt accgcgtcga gaagttccta ttccgaagtt cctattctct agaaagtata
     4141 ggaacttcgg gcccgcggtg gcggcgatct ctccccagca tgcctgctat tgtcttccca
     4201 atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag
     4261 acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
     4321 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
     4381 cgaggctgat cagcgggccg ctttacttgt acagctcgtc catgccgccg gtggagtggc
     4441 ggccctcggc gcgttcgtac tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt
     4501 ccaacttgat gttgacgttg taggcgccgg gcagctgcac gggcttcttg gccttgtagg
     4561 tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag cttcagcctc tgcttgatct
     4621 cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga ggaggcctcc cagcccatgg
     4681 tcttcttctg cattacgggg ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt
     4741 agatgaactc gccgtcctgc agggaggagt cctgggtcac ggtcaccacg ccgccgtcct
     4801 cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc aagtagtcgg
     4861 ggatgtcggc ggggtgcttc acgtaggcct tggagccgta catgaactga ggggacagga
     4921 tgtcccaggc gaagggcagg gggccaccct tggtcacctt cagcttggcg gtctgggtgc
     4981 cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc
     5041 cctccatgtg caccttgaag cgcatgaact ccttgatgat ggccatgtta tcctcctcgc
     5101 ccttgctcac catgttgtca gacctggtaa ctccttactg taacttattg cacctgatga
     5161 acaaaagatg tgaactgaac gcaagtagga ggctaataga ggctagtaca gtacaaagat
     5221 ccccaaatga tgtgtgccgg ccttagtgga ggcagacccc tggttatata cctgtgctgc
     5281 agaaagaggg gcttacaaat ggatctctct ggaatggcat tatctcatgg tgggaacagt
     5341 gttgtcagca gaatcagcgg ggtggactct gacctgagaa tgacagtcta gcactgaatg
     5401 gctcagacaa tactcttgtg gccaactatc accggaaaat aagtgcctgc caaaacaaac
     5461 cagaccagat gaagctacat ggctgtagaa cgtcagtatt taatgtgttg tggccacttg
     5521 gttcaccaga tttaagagct aattttttta aatatcacag ataatcaatc gctttttttt
     5581 tttcttgact gaaaagctta gactggaaaa ttctttgaaa aagtttgcac tacagaaaat
     5641 aaatacataa catcctgcac tgaatgcaca ctattaatga agttcctata ctttctagag
     5701 aataggaact tcggtaccca gcttttgttc cctttagtga gggttaattg cgcgctaggg
     5761 ataacagggt aatacgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5821 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5881 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgkga
     5941 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6001 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6061 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6121 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6181 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6241 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6301 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6361 cgcgaatttt aacaaaatat taacgcttac aaktyaggtg gcacttttcg gggaaatgtg
     6421 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6481 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6541 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6601 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6661 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6721 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6781 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6841 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6901 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6961 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7021 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7081 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7141 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7201 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7261 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7321 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7381 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7441 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7501 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7681 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7921 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8161 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8221 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8281 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8341 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8401 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8461 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8521 caatttcaca caggaaacag ctatgaccat
//
