LOCUS       sequence-267553-       10434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for constitutive expression of Afamin + Wnt3a for 
            production of conditioned media in HEK293T cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10434)
  AUTHORS   Fischer AD, Veronese-Paniagua DA, Swaminathan S, Kashima H, Rubin 
            DC, Madison BB
  TITLE     The oncogenic function of PLAGL2 is mediated via ASCL2 and IGF2 and 
            a Wnt-independent mechanism in colorectal cancer.
  JOURNAL   Am J Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022.
  PUBMED    37310750
REFERENCE   2  (bases 1 to 10434)
  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: "Am J 
            Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022."
COMMENT     Sequence Label: BII-CMV-AfmW3A
FEATURES             Location/Qualifiers
     source          1..10434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   570..604
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(912..928)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(912..928)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     enhancer        1237..1540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1541..1744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1695..1715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1741..1765
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3667..3720
                     /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"
     misc_feature    4821..5393
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4988..5005)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5215..5234
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5396..5998
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5895..5915
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6025..6613
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6078..6098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6496..6507)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6643..6777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6667..6686)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6721..6740
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7556..7618
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7748..7765)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7748..7764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7757..7779)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7906..8361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7993..8012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8203..8224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8387..8491
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8492..8560,8561..9352)
                     /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(8710..8729)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9523..10111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10012..10031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10265..10282
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10399..10420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagcgcgcc cgccgggtaa ctcacggggt
      121 atccatgtcc atttctgcgg catccagcca ggatacccgt cctcgctgac gtaatatccc
      181 agcgccgcac cgctgtcatt aatctgcaca ccggcacggc agttccggct gtcgccggta
      241 ttgttcgggt tgctgatgcg cttcgggctg accatccgga actgtgtccg gaaaagccgc
      301 gacgaactgg tatcccaggt ggcctgaacg aacagttcac cgttaaaggc gtgcatggcc
      361 acaccttccc gaatcatcat ggtaaacgtg cgttttcgct caacgtcaat gcagcagcag
      421 tcatcctcgg caaactcttt ccatgccgct tcaacctcgc gggaaaaggc acgggcttct
      481 tcctccccga tgcccagata gcgccagctt gggcgatgac tgagccggaa aaaagacccg
      541 acgatatgat cctgatgcag ctagattaac cctagaaaga tagtctgcgt aaaattgacg
      601 catgcattct tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt
      661 taggacatct cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg
      721 tcactgattt tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg
      781 tgacttttaa gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg
      841 ataacttatt atatatatat tttcttgtta tagatctgtg gaatgtgtgc tagcattagt
      901 tattaatagg ggatccacta gttctagagg gacagccccc ccccaaagcc cccagggatg
      961 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
     1021 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
     1081 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
     1141 ggggggatac ggggaaaagg cctccaaggc ctgcaggagt tattaatagt aatcaattac
     1201 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1261 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1321 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1381 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     1441 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     1501 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1561 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1621 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     1681 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     1741 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     1801 tagaagacac ccactagtct ggccaccatg agacacttaa aacttacagg ttttattttt
     1861 ttcctgttgc ctttgactga atccctagcc ctacccacaa agcctcagga cgtagatcat
     1921 ttcaatgcta cccagaaatt catagatgag aacacgacat acctggccat catcgcgttt
     1981 tcccagtatg tccaggaagc atcctttgat gaggtggaaa cgctggtgaa agtcatgctg
     2041 gattacaggg ataggtgctg ggctgacaac acacttccag agtgttcaaa gacagctaat
     2101 gacgctatcc aggacatgtt atgtgacatg gagggactgc cgcagaagca caacttttct
     2161 cattgctgcg ggaaagccgg ctttccaaga agactctgtt tcttctacaa caagaaagct
     2221 aatgtaggat tcttgccccc tttccccact ctggatccag aagagaaatg ccaagcttat
     2281 aaaaataaca gtgaatcttt tctacacctc tatatgtatg aggttgccag aaggaacccc
     2341 tttgtcttcg cccccgttct tctagccgtg gctgcttggt ttgaggaggc agcgaccaca
     2401 tgttgtgaac agcaacagaa agccacctgc tttcaagcca aggcagctcc catcacacaa
     2461 tatttaaaag catcatcttc ttatcaaaga aatgtttgtg gagcccttat aaagtttgga
     2521 cccaaggtct taaactctat aaacgttgct gtattcagta aaaaattccc caagattgga
     2581 tttaaggacc ttaccaccct cttagaagat gtttcttcca tgtatgaagg atgctgtgaa
     2641 ggagatgttg tgcactgcat ccgaagccag agccaggttg tgaaccatat ttgttcaaaa
     2701 caagattcca tctcaagtaa aatcaaagtg tgctgcgaaa agaagaccct agaacgtgaa
     2761 gcctgtataa ttaatgcaaa taaagacgac agaccggagg gcttatctct cagagaagcc
     2821 aaattcactg agagcgaaaa tgtgtgtcaa gaacgagatt ctgacccgga caagttcttt
     2881 gctgagttta tttatgaata ctccaggaga catcctgacc tgtctacacc agagctccta
     2941 agaattacta aagtgtatat ggacttctta gaagactgct gcagcagaga aaacccagca
     3001 ggttgttacc gccatgtgga ggataaattc aatgagacaa ctcagcgaag ccttgcaatg
     3061 gtgcagcagg agtgcaaaca gttccaggag ctggggaagg acactctgca gcgccatttc
     3121 cttgtcaagt tcaccaaggc agcccctcag ctccccatgg aagaactggt ctcccttagt
     3181 aaggaaatgg tggcagctct gacaacctgt tgcactctca gtgacgagtt cgcctgcgtt
     3241 gacaacttgg cagatttagt tcttggagag ttatgtgggg taaatacaaa tcgaactatc
     3301 aatcctgctg tggatcactg ctgcaaaaca gacttcgcct tcagaagaca ctgctttgag
     3361 catctgaaag ctgatacaac gtatgagctc ccatcagtct ccgcacttgt atctgctttg
     3421 catacagact ggtgtcagcc tcgtaaggag gaccttcaga ataagaaaca caggtttctg
     3481 gtcaacttgg tgaagtggat gccagggatc acggatgagg agtggctgtg cttgttcaca
     3541 aagttcactg ctgccaggga ggagtgcagc gaagtgcagg agcctgagtc ctgtttcagt
     3601 ccagagagtt caaaaactgg tgacgaaagt caagctacag agaaacaaag aagccttggc
     3661 tccggcgagg gcaggggaag tcttctaaca tgcggggacg tggaggaaaa tcccggccca
     3721 ctcggcatgg ccccactcgg atacttctta ctcctctgca gcctgaagca ggctctgggc
     3781 agctacccga tctggtggtc gctggctgtt gggccacagt attcctccct gggctcgcag
     3841 cccatcctgt gtgccagcat cccgggcctg gtccccaagc agctccgctt ctgcaggaac
     3901 tacgtggaga tcatgcccag cgtggccgag ggcatcaaga ttggcatcca ggagtgccag
     3961 caccagttcc gcggccgccg gtggaactgc accaccgtcc acgacagcct ggccatcttc
     4021 gggcccgtgc tggacaaagc taccagggag tcggcctttg tccacgccat tgcctcagcc
     4081 ggtgtggcct ttgcagtgac acgctcatgt gcagaaggca cggccgccat ctgtggctgc
     4141 agcagccgcc accagggctc accaggcaag ggctggaagt ggggtggctg tagcgaggac
     4201 atcgagtttg gtgggatggt gtctcgggag ttcgccgacg cccgggagaa ccggccagat
     4261 gcccgctcag ccatgaaccg ccacaacaac gaggctgggc gccaggccat cgccagccac
     4321 atgcacctca agtgcaagtg ccacgggctg tcgggcagct gcgaggtgaa gacatgctgg
     4381 tggtcgcaac ccgacttccg cgccatcggt gacttcctca aggacaagta cgacagcgcc
     4441 tcggagatgg tggtggagaa gcaccgggag tcccgcggct gggtggagac cctgcggccg
     4501 cgctacacct acttcaaggt gcccacggag cgcgacctgg tctactacga ggcctcgccc
     4561 aacttctgcg agcccaaccc tgagacgggc tccttcggca cgcgcgaccg cacctgcaac
     4621 gtcagctcgc acggcatcga cggctgcgac ctgctgtgct gcggccgcgg ccacaacgcg
     4681 cgagcggagc ggcgccggga gaagtgccgc tgcgtgttcc actggtgctg ctacgtcagc
     4741 tgccaggagt gcacgcgcgt ctacgacgtg cacacctgca agtgaccact gaaatggaca
     4801 agtagcggcc gcaaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     4861 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     4921 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     4981 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     5041 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     5101 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     5161 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     5221 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     5281 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     5341 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggc
     5401 caccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     5461 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     5521 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     5581 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     5641 cgtcgaagcg ggggcggtgt tcgccgagat cggctcgcgc atggccgagt tgagcggttc
     5701 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     5761 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5821 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgct ggggtgcccg ccttcctgga
     5881 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     5941 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt
     6001 ttgtttgaat gacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg
     6061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     6121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6181 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6241 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6301 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6361 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     6421 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6481 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6541 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6601 gccgcctccc cgcctggaat taattctgca atcgatacta gtgatccaga catgataaga
     6661 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     6721 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     6781 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     6841 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga gggacagccc
     6901 ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc agcagcgagc
     6961 cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg
     7021 ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg cttctcgctg
     7081 ctctttgagc ctgcagacac ctggggggat acggggaaaa agccatacca atggagctca
     7141 cggggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
     7201 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
     7261 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
     7321 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaagaattcg
     7381 ataaaagttt tgttacttta tagaagaaat tttgagtttt tgtttttttt aataaataaa
     7441 taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa atataataaa
     7501 acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata aaacacatgc
     7561 gtcaatttta cacatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt
     7621 aatctagctg cgtgttctgc agcgtgtcga gcatcttcat ctgctccatc acgctgtaaa
     7681 acacatttgc accgcgagtc tgcccgtcct ccacgggttc aaaaacgtga atgaacgagg
     7741 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     7801 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     7861 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     7921 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     7981 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     8041 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     8101 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     8161 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     8221 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     8281 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     8341 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     8401 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     8461 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     8521 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     8581 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     8641 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     8701 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     8761 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     8821 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     8881 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     8941 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     9001 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     9061 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     9121 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     9181 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     9241 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     9301 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     9361 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     9421 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     9481 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     9541 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     9601 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     9661 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     9721 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     9781 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     9841 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     9901 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     9961 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
    10021 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
    10081 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
    10141 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
    10201 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
    10261 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
    10321 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
    10381 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggc
//
