LOCUS       sequence-413074-        9825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector backbone encoding doxycycline-inducible expression 
            of membrane-localized Akt1 (without pleckstrin homology domain).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9825)
  AUTHORS   Underhill EJ, Toettcher JE
  TITLE     Control of gastruloid patterning and morphogenesis by the Erk and 
            Akt signaling pathways.
  JOURNAL   Development. 2023 Aug 15;150(16):dev201663. doi: 10.1242/dev.201663.
            Epub 2023 Aug 17.
  PUBMED    37590131
REFERENCE   2  (bases 1 to 9825)
  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: 
            "Development."; date: "2023-08-15"; volume: "150(16):dev201663. 
            doi"; pages: " 10.1242/dev.201663. Epub 2023 Aug 17"
COMMENT     Sequence Label: Dox-Akt
FEATURES             Location/Qualifiers
     source          1..9825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             75..770
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     CDS             780..836
                     /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             837..1583
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    1688..2276
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2159..2170)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   2519..2581
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2693..2712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2812..2834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2872..2890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2958..3062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3063..3131,3132..3923)
                     /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(3281..3300)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4094..4682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4583..4602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4836..4853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4970..4991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5006..5011,5012..5029,5030..5036)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5044..5060
                     /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     5049..5071
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5068..5084
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5068..5084
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5119..5153
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        5534..5912
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    5542..5560
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5578..5596
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5614..5632
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5650..5668
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5686..5704
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5722..5740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5758..5776
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5821..5845
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5958..5982
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5988..6008
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             6036..6734
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             7818..7844
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7824..7850
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     misc_feature    7970..8558
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(8441..8452)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    8803..8821
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8824..8842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          8887..9825
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     9782..9802
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gtgtcgtgag gggatctgga gctctcgaga attctcacgc gtcaagtgga gcaaggcagg
       61 tggacagtgg atccatggtg agcgagctga ttaaggagaa catgcccatg aagctgtaca
      121 tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct
      181 acgagggcac ccagaccatg agaatcaagg tcgtcgaggg cggccctctc cccttcgcct
      241 tcgacatcct ggctaccagc ttcatgtacg gcagcagaac cttcatcaag caccctccgg
      301 gcatccccga cttctttaag cagtccttcc ctgagggctt cacatgggag agagtcacca
      361 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
      421 tcatctacaa cgtcaaggtt agaggggtga acttcccagc caacggccct gtgatgcaga
      481 agaaaacact cggctgggag gcctccaccg agacgatgta ccccgctgac ggcggcctgg
      541 aaggcgcatg tgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaaccttg
      601 agaccacata cagatccaag aaacccgcta cgaacctcaa gatgcccggc gtctacaacg
      661 tggaccacag actggaaaga atcaaggagg ccgacgatga gacctacgtc gagcagcacg
      721 aggtggctgt ggccagatac tctactggtg gcgctggtga tggaggtaaa ggaagcggag
      781 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctatga
      841 gccggctgga caagagcaaa gtgatcaaca gcgccctgga actgctgaac ggcgtgggca
      901 tcgagggcct gaccacccgg aagctggccc agaaactggg cgtggaacag cccaccctgt
      961 actggcacgt gaagaacaag cgggccctgc tggacgccct gcccatcgag atgctggacc
     1021 ggcaccacac ccacagctgc cctctggaag gcgagagctg gcaggacttc ctgcggaaca
     1081 acgccaagag ctacagatgc gccctgctga gccaccggga cggcgccaaa gtgcacctgg
     1141 gcaccagacc caccgagaag cagtacgaga cactggaaaa ccagctggcc ttcctgtgcc
     1201 agcagggctt cagcctggaa aacgccctgt acgccctgag cgccgtgggc cacttcaccc
     1261 tgggctgcgt gctggaagaa caggaacacc aggtcgccaa agaggaaaga gagacaccca
     1321 ccaccgacag catgcccccc ctgctgaagc aggccatcga gctgttcgac agacagggcg
     1381 ccgagcccgc cttcctgttc ggcctggaac tgatcatctg cggcctcgag aagcagctga
     1441 agtgcgagag cggcggaccc accgacgccc tggacgactt cgacctggac atgctgcccg
     1501 ccgatgccct ggatgatttt gatctggata tgctccctgc tgacgctctc gatgacttcg
     1561 atctcgatat gctgccaggc tagcctgcag gcggccgcaa ctcccacctg caacatgcgt
     1621 gactgactga ggccgcgact ctagagtcga cctgcaggca tgcaagcttg atatcaagct
     1681 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1741 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1801 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1861 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1921 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1981 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2041 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2101 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2161 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2221 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     2281 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgatact
     2341 agttaaaagt tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat
     2401 aaataaacat aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat
     2461 aaaacttaat atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca
     2521 tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg
     2581 gttaatctag tatacgcgta tgcggccgct taattaatcc atggcgcctg atgcggtatt
     2641 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     2701 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2761 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2821 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     2881 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2941 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3001 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3061 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3121 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3181 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3241 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3301 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3361 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3421 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3481 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3541 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3601 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3661 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3721 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3781 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3841 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3901 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3961 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4021 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4201 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4261 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4321 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4381 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4441 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4501 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4561 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4621 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4681 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4741 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4801 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4861 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4921 ggcacgacag gtttcccgac tggaaagcgg gcagtgatcg caacgcaatt aatgtgagtt
     4981 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5041 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccgggg
     5101 cgcgccattc tagattaacc ctagaaagat agtctgcgta aaattgacgc atgcattctt
     5161 gaaatattgc tctctctttc taaatagcgc gaatccgtcg ctgtgcattt aggacatctc
     5221 agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt
     5281 gaactataac gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt gacttttaag
     5341 atttaactca tacgataatt atattgttat ttcatgttct acttacgtga taacttatta
     5401 tatatatatt ttcttgttat agatatcaac tagaatgcta gcatgggccc agatctccga
     5461 ggggacccga caggcccgca gggacagcag agatccagtt tggatcgatc gatcacgaga
     5521 ctagcctcga gttggcttta ctccctatca gtgatagaga acgtatgaag agtttactcc
     5581 ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa
     5641 ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata
     5701 gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc
     5761 ctatcagtga tagagaacgt ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag
     5821 agctcgttta gtgaaccgtc agatcgcctg gagcaattcc acaacacttt tgtcttatac
     5881 caactttccg taccacttcc taccctcgta aaccggttcg tcgactagtc cagtgtggtg
     5941 gaattctgca gatatcaaca agtttgtaca aaaaagcagg ctccaccatg ggttcctcca
     6001 aatccaagcc caaggacgca tctcagcgac gtcgcatgag cgagctgatt aaggagaaca
     6061 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     6121 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     6181 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     6241 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     6301 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     6361 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     6421 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc
     6481 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     6541 atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga
     6601 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
     6661 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     6721 ggcacaagct taatagtgct ggtggtgaat tcatgagcga ggctgaagag atggaggtgt
     6781 ccctggccaa gcccaagcac cgcgtgacca tgaacgagtt tgagtacctg aagctgctgg
     6841 gcaagggcac tttcggcaag gtgatcctgg tgaaggagaa ggccacaggc cgctactacg
     6901 ccatgaagat cctcaagaag gaagtcatcg tggccaagga cgaggtggcc cacacactca
     6961 ccgagaaccg cgtcctgcag aactccaggc accccttcct cacagccctg aagtactctt
     7021 tccagaccca cgaccgcctc tgctttgtca tggagtacgc caacgggggc gagctgttct
     7081 tccacctgtc ccgggaacgt gtgttctccg aggaccgggc ccgcttctat ggcgctgaga
     7141 ttgtgtcagc cctggactac ctgcactcgg agaagaacgt ggtgtaccgg gacctcaagc
     7201 tggagaacct catgctggac aaggacgggc acattaagat cacagacttc gggctgtgca
     7261 aggaggggat caaggacggt gccaccatga agaccttttg cggcacacct gagtacctgg
     7321 cccccgaggt gctggaggac aatgactacg gccgtgcagt ggactggtgg gggctgggcg
     7381 tggtcatgta cgagatgatg tgcggtcgcc tgcccttcta caaccaggac catgagaagc
     7441 tttttgagct catcctcatg gaggagatcc gcttcccgcg cacgcttggt cccgaggcca
     7501 agtccttgct ttcagggctg ctcaagaagg accccaagca gaggcttggc gggggctccg
     7561 aggacgccaa ggagatcatg cagcatcgct tctttgccgg tatcgtgtgg cagcacgtgt
     7621 acgagaagaa gctcagccca cccttcaagc cccaggtcac gtcggagact gacaccaggt
     7681 attttgatga ggagttcacg gcccagatga tcaccatcac accacctgac caagatgaca
     7741 gcatggagtg tgtggacagc gagcgcaggc cccacttccc ccagttctcc tactcggcca
     7801 gcagcacggc catggcttat ccttacgacg tccccgacta cgccagtctg ggacctggac
     7861 tgtgacctgc aggcggccgc aactcccacc tgcaacatgc gtgactgact gaggccgcga
     7921 ctctagagtc gacctgcagg catgcaagct tgatatcaag cttatcgata atcaacctct
     7981 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     8041 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     8101 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     8161 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     8221 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     8281 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     8341 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     8401 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     8461 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     8521 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgcccgcccc acgacccgca
     8581 gcgcccgacc gaaaggagcg cacgacccca tgcatcgggc tccggtgccc gtcagtgggc
     8641 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
     8701 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     8761 ttttcccgag ggtgggggag aaccgtatat aagtgcagga tctccctatc agtgatagag
     8821 atctccctat cagtgataga gatcgtcgac gttctttttc gcaacgggtt tgccgccaga
     8881 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
     8941 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
     9001 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
     9061 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
     9121 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
     9181 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
     9241 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
     9301 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
     9361 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     9421 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     9481 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     9541 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     9601 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     9661 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     9721 tggcacttga tgtaattctc cttggaattt gcccttttta agtttggatc ttggttcatt
     9781 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcag
//
