LOCUS       sequence-386851-       10211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To establish Ngn3ER cell line for endocrine differentiation 
            (4-hydroxytamoxifen activation, Puromycin resistant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10211)
  AUTHORS   Huang X, Gu W, Zhang J, Lan Y, Colarusso JL, Li S, Pertl C, Lu J, 
            Kim H, Zhu J, Breault DT, Sevigny J, Zhou Q
  TITLE     Stomach-derived human insulin-secreting organoids restore glucose 
            homeostasis.
  JOURNAL   Nat Cell Biol. 2023 Apr 27. doi: 10.1038/s41556-023-01130-y.
  PUBMED    37106062
REFERENCE   2  (bases 1 to 10211)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2023 Apr 27. doi: 10.1038/s41556-023-01130-y."
COMMENT     Sequence Label: pLenti-EF1a-Ngn3ER-PuroR-mCherry
FEATURES             Location/Qualifiers
     source          1..10211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    351..372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(387..392,393..410,411..417)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    425..441
                     /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     430..452
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     449..465
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     449..465
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     515..534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             726..1359
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1406..1531
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2028..2261
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2446..2490
                     /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    2783..2900
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2948..4126
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3179..4117
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4074..4094
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    4139..4163
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4191..4200
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5811..5867
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5874..6470
                     /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(5874..5893)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6370..6390
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6477..6530
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6537..7247
                     /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(6682..6700)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6950..6969
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7254..7313
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     protein_bind    complement(7337..7361)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    7378..7966
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7431..7451)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7849..7860)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8114..8294
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8341..8359)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8427..8531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8532..8600,8601..9392)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8750..8769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9563..10151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag
      181 ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
      301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      361 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      421 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc
      481 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
      541 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
      601 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
      661 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
      721 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
      781 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
      841 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
      901 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
      961 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
     1021 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
     1081 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
     1141 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
     1201 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1261 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     1321 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     1381 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     1441 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     1501 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     1561 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     1621 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1681 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1741 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1801 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1861 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1921 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1981 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     2041 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     2101 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     2161 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     2221 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     2281 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     2341 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     2401 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     2461 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     2521 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     2581 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     2641 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     2701 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg ccgaattcac
     2761 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2821 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2881 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggacta
     2941 gtcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     3001 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
     3061 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     3121 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
     3181 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
     3241 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     3301 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     3361 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     3421 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     3481 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     3541 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     3601 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     3661 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     3721 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     3781 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     3841 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     3901 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     3961 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     4021 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     4081 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaagcg gccatcaaac
     4141 aagtttgtac aaaaaagcag gctggcgccg gaaccaattc agtcgacgcc gccaccatgg
     4201 cgcctcatcc cttggatgcg ctcaccatcc aagtgtcccc agagacacaa caaccttttc
     4261 ccggagcctc ggaccacgaa gtgctcagtt ccaattccac cccacctagc cccactctca
     4321 tacctaggga ctgctccgaa gcagaagtgg gtgactgccg agggacctcg aggaagctcc
     4381 gcgcccgacg cggagggcgc aacaggccca agagcgagtt ggcactcagc aaacagcgaa
     4441 gaagccggcg caagaaggcc aatgatcggg agcgcaatcg catgcacaac ctcaactcgg
     4501 cgctggatgc gctgcgcggt gtcctgccca ccttcccgga tgacgccaaa cttacaaaga
     4561 tcgagaccct gcgcttcgcc cacaactaca tctgggcact gactcagacg ctgcgcatag
     4621 cggaccacag cttctatggc ccggagcccc ctgtgccctg tggagagctg gggagccccg
     4681 gaggtggctc caacggggac tggggctcta tctactcccc agtctcccaa gcgggtaacc
     4741 tgagccccac ggcctcattg gaggaattcc ctggcctgca ggtgcccagc tccccatcct
     4801 atctgctccc gggagcactg gtgttctcag acttcttgag ggatccacga aatgaaatgg
     4861 gtgcttcagg agacatgagg gctgccaacc tttggccaag ccctcttgtg attaagcaca
     4921 ctaagaagaa tagccctgcc ttgtccttga cagctgacca gatggtcagt gccttgttgg
     4981 atgctgaacc gcccatgatc tattctgaat atgatccttc tagacccttc agtgaagcct
     5041 caatgatggg cttattgacc aacctagcag atagggagct ggttcatatg atcaactggg
     5101 caaagagagt gccaggcttt ggggacttga atctccatga tcaggtccac cttctcgagt
     5161 gtgcctggct ggagattctg atgattggtc tcgtctggcg ctccatggaa cacccgggga
     5221 agctcctgtt tgctcctaac ttgctcctgg acaggaatca aggtaaatgt gtggaaggca
     5281 tggtggagat ctttgacatg ttgcttgcta cgtcaagtcg gttccgcatg atgaacctgc
     5341 agggtgaaga gtttgtgtgc ctcaaatcca tcattttgct taattccgga gtgtacacgt
     5401 ttctgtccag caccttgaag tctctggaag agaaggacca catccaccgt gtcctggaca
     5461 agatcacaga cactttgatc cacctgatgg ccaaagctgg cctgactctg cagcagcagc
     5521 atcgccgcct agctcagctc cttctcattc tttcccatat ccggcatatg agtaacaaac
     5581 gcatggagca tctctacaac atgaaatgca agaacgtggt acccctctat gacctgctcc
     5641 tggagatgtt ggatgcccac cgccttcatg ccccagccag tcgcatggga gtgcccccag
     5701 aggagcccag ccagacccag ctggccacca ccagctccac ttcagcacat tccttacaaa
     5761 cctactacat acccccggaa gcagagggct tccccaacac gatcactagt gccacgaact
     5821 tctctctgtt aaagcaagca ggagatgttg aagaaaaccc cgggcctgga tccatgaccg
     5881 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     5941 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     6001 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     6061 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     6121 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     6181 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     6241 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     6301 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     6361 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     6421 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc ggatccgagg
     6481 gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct atcgatatgg
     6541 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     6601 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     6661 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     6721 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     6781 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     6841 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     6901 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     6961 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     7021 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     7081 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     7141 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     7201 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaatc gatcagtgta
     7261 ctaattatgc tctcttgaaa ttggctggag atgttgagag caacccaggt cccgcggccg
     7321 cactcgagat atctagaccc agctttcttg tacaaagtgg ttcgatcgat agatcctaat
     7381 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     7441 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     7501 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     7561 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     7621 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7681 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     7741 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     7801 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     7861 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     7921 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag
     7981 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     8041 ggaagggcta attcactccc aacgaagaca agatcacctg caggacaggc gcgccctgct
     8101 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     8161 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     8221 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     8281 gaaaatctct agcacccggg cgattaagga aagggctaga tcattcttga agacgaaagg
     8341 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     8401 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     8461 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     8521 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     8581 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     8641 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     8701 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     8761 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     8821 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     8881 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     8941 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     9001 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     9061 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     9121 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     9181 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     9241 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     9301 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     9361 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     9421 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     9481 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     9541 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     9601 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     9661 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     9721 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     9781 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     9841 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     9901 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     9961 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
    10021 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
    10081 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
    10141 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
    10201 ttgctcacat g
//
