LOCUS       sequence-247972-       11240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for doxycycline-inducible BMP7 expression from the murine 
            ROSA26 safe harbor locus upon CRISPR/Cas9-mediated genomic insertion
            and stable selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11240)
  AUTHORS   Routh ED, Pullikuth AK, Jin G, Chifman J, Chou JW, D'Agostino RB Jr,
            Seino KI, Wada H, Print CG, Zhang W, Lu Y, Miller LD
  TITLE     Transcriptomic Features of T Cell-Barren Tumors Are Conserved Across
            Diverse Tumor Types.
  JOURNAL   Front Immunol. 2020 Feb 13;11:57. doi: 10.3389/fimmu.2020.00057. 
            eCollection 2020.
  PUBMED    32117236
REFERENCE   2  (bases 1 to 11240)
  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: "Front 
            Immunol."; date: "2020-02-13"; volume: "11:57. doi"; pages: " 
            10.3389/fimmu.2020.00057. eCollection 2020"
COMMENT     Sequence Label: pR26-mBMP7
FEATURES             Location/Qualifiers
     source          1..11240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..794
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    join(813..878,879..916)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    931..979
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        1004..1318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1012..1030
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1048..1066
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1083..1101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1119..1137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1155..1173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1190..1208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1226..1244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1291..1315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    2674..2722
                     /note="synthetic polyadenylation signal"
     promoter        2736..3013
                     /label=chicken beta-actin promoter
     intron          3316..3888
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     3896..3915
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        3943..3961
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3985..4602,4603..4608,4609..4644,4645..4647,4648..4683,
                     4684..4686,4687..4722,4723..4731)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGAR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    4734..4959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6002..6021
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6180..6779
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVESAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6180..6199)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6676..6696
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6780..6901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6817..6836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6871..6890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    6958..7768
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     promoter        complement(7843..7861)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7868..7885)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7868..7884)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7877..7899)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8022..8324
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     8203..8222
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             8673..9467
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             8673..9467
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8727..8746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9337..9356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      9667..9676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9673..10047
                     /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"
     rep_origin      10185..10773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10674..10693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10927..10944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11061..11082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11097..11102,11103..11120,11121..11127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11135..11151
                     /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     11140..11162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11159..11175
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11159..11175
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        11193..11211
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     11193..11210
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cagttaacgg cagccggagt gcgcagccgc cggcagcctc gctctgccca ctgggtgggg
       61 cgggaggtag gtggggtgag gcgagctgga cgtgcgggcg cggtcggcct ctggcggggc
      121 gggggggggg ggggrgggtc agcgaaagta gctcgcgcgc gagcggccgc ccaccctccc
      181 cttcctctgg gggagtcgtt ttacccgccg ccggccgggc ctcgtcgtct gattggctct
      241 cggggcccag aaaactggcc cttgccattg gctcgtgttc gtgcaagttg agtccatccg
      301 ccggccagcg ggggcggcga ggaggcgctc ccaggttccg gccctcccct cggccccgcg
      361 ccgcagagtc tggccgcgcg cccctgcgca acgtggcagg aagcgcgcgc tgggggcggg
      421 gacgggcagt agggctgagc ggctgcgggg cgggtgcaag cacgtttccg acttgagttg
      481 cctcaagagg ggcgtgctga gccagacctc catcgcgcac tccggggagt ggagggaagg
      541 agcgagggct cagttgggct gttttggagg caggaagcac ttgctctccc aaagtcgctc
      601 tgagttgtta tcagtaaggg agctgcagtg gagtaggcgg ggagaaggcc gcacccttct
      661 ccggaggggg gaggggagtg ttgcaatacc tttctgggag ttctctgctg cctcctggct
      721 tctgaggacc gccctgggcc tgggagaatc ccttccccct cttccctcgt gatctgcaac
      781 tccagtcttt ctagaatcga tctcgagaac tgtagggcgc agtagtccag ggtttccttg
      841 atgatgtcat acttatcctg tccctttttt ttccacagct cgcggttgag gacaaactct
      901 tcgcggtctt tccagtggtt aattagggat cggcaataaa aagacagaat aaaacgcacg
      961 ggtgttgggt cgtttgttca tcgatgccct ttcgtcttca ttcgagttta ctccctatca
     1021 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
     1081 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     1141 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
     1201 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
     1261 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     1321 gagaattcga gctcggtacc cggggatgct agcgaggaga tctgccacca tgcacgtgcg
     1381 ctcgctgcgc gctgcggcgc cacacagctt cgtggcgctc tgggcgcctc tgttcttgct
     1441 gcgctccgcc ctggccgatt tcagcctgga caacgaggtg cactccagct tcatccaccg
     1501 gcgcctccgc agccaggagc ggcgggagat gcagcgggag atcctgtcca tcttagggtt
     1561 gccccatcgc ccgcgcccgc acctccaggg aaagcataat tcggcgccca tgttcatgtt
     1621 ggacctgtac aacgccatgg cggtggagga gagcgggccg gacggacagg gcttctccta
     1681 cccctacaag gccgtcttca gtacccaggg ccccccttta gccagcctgc aggacagcca
     1741 cttcctcact gacgccgaca tggtcatgag cttcgtcaac ctagtggaac atgacaaaga
     1801 attcttccac cctcgatacc accatcggga gttccggttt gatctttcca agatccccga
     1861 gggcgaagcg gtgaccgcag ccgaattcag gatctataag gactacatcc gggagcgatt
     1921 tgacaacgag accttccaga tcacagtcta tcaggtgctc caggagcact caggcaggga
     1981 gtcggacctc ttcttgctgg acagccgcac catctgggct tctgaggagg gctggttggt
     2041 gtttgatatc acagccacca gcaaccactg ggtggtcaac cctcggcaca acctgggctt
     2101 acagctctct gtggagaccc tggatgggca gagcatcaac cccaagttgg caggcctgat
     2161 tggacggcat ggaccccaga acaagcaacc cttcatggtg gccttcttca aggccacgga
     2221 agtccatctc cgtagtatcc ggtccacggg gggcaagcag cgcagccaga atcgctccaa
     2281 gacgccaaag aaccaagagg ccctgaggat ggccagtgtg gcagaaaaca gcagcagtga
     2341 ccagaggcag gcctgcaaga aacatgagct gtacgtcagc ttccgagacc ttggctggca
     2401 ggactggatc attgcacctg aaggctatgc tgcctactac tgtgagggag agtgcgcctt
     2461 ccctctgaac tcctacatga acgccaccaa ccacgccatc gtccagacac tggttcactt
     2521 catcaaccca gacacagtac ccaagccctg ctgtgcgccc acccagctca acgccatctc
     2581 tgtcctctac ttcgacgaca gctctaatgt catcctgaag aagtacagaa acatggtggt
     2641 ccgggcctgt ggctgccact agacgcgtta ggaaataaaa gatctttatt ttcattagat
     2701 ctgtgtgttg gttttttgtg tgccgcgtcc tagagtcgag gtgagcccca cgttctgctt
     2761 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     2821 attttgtgca gcgatggggg cgcggggggg gggggggcgc gcgccaggcg gggcgggggg
     2881 gggcgggggg gggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     2941 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     3001 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     3061 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     3121 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
     3181 gtggctgcgt gaaagccttg aggggctccg ggaggatctt ctagggcctc tgagctattc
     3241 cagaagtagt gaagaggctt ttttggaggc ctaggctttt gcaaaaagct ccggatcgat
     3301 cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta
     3361 aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt
     3421 cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga
     3481 caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc
     3541 ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt
     3601 ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt
     3661 tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc
     3721 tggcgtggaa atattcttat tggtagaaac aactacatcc tggtcatcat cctgcctttc
     3781 tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa
     3841 ccgggcccct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     3901 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgtaatacga ctcactatag
     3961 ggcgaattga tcggatccgc caccatgtcc agactggaca agagcaaagt cataaacggc
     4021 gctctggaat tactcaatgg agtcggtatc gaaggcctga cgacaaggaa actcgctcaa
     4081 aagctgggag ttgagcagcc taccctgtac tggcacgtga agaacaagcg ggccctgctc
     4141 gatgccctgc caatcgagat gctggacagg catcataccc acttctgccc cctggaaggc
     4201 gagtcatggc aagactttct gcggaacaac gccaagtcat tccgctgtgc tctcctctca
     4261 catcgcgacg gggctaaagt gcatctcggc gcccgcccaa cagagaaaca gtacgaaacc
     4321 ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct ccctggagaa cgcactgtac
     4381 gctctgtccg ccgtgggcca ctttacactg ggctgcgtat tggaggaaca ggagcatcaa
     4441 gtagcaaaag aggaaagaga gacacctacc accgattcta tgcccccact tctgagacaa
     4501 gcaattgagc tgttcgaccg gcagggagcc gaacctgcct tccttttcgg cctggaacta
     4561 atcatatgtg gcctggagaa acagctaaag tgcgaaagcg gcgggccggc cgacgccctt
     4621 gacgattttg acttagacat gctcccagcc gatgcccttg acgactttga ccttgatatg
     4681 ctgcctgctg acgctcttga cgattttgac cttgacatgc tccccgggta aggctgtgcc
     4741 tttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4801 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4861 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4921 acaatagcag gcatgctggg gatgcggtgg gctctatggc gcgccgggtt ttggcgcctc
     4981 ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc
     5041 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     5101 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     5161 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     5221 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     5281 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     5341 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     5401 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     5461 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     5521 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     5581 gtgggcggca agaacccaag gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg
     5641 gtgagatggg ctggggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     5701 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     5761 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     5821 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     5881 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     5941 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     6001 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     6061 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt
     6121 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagaccaagt ttggccacca
     6181 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     6241 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     6301 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6361 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6421 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     6481 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6541 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     6601 gcgccgtcgt gctccccgga gtggagtcgg ccgagcgcgc cggggtgccc gccttcctgg
     6661 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6721 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa
     6781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6901 agggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt cgcccttaga
     6961 tgggcgggag tcttctgggc aggcttaaag gctaacctgg tgtgtgggcg ttgtcctgca
     7021 ggggaattga acaggtgtaa aattggaggg acaagacttc ccacagattt tcggttttgt
     7081 cgggaagttt tttaataggg gcaaataagg aaaatgggag gataggtagt catctggggt
     7141 tttatgcagc aaaactacag gttattattg cttgtgatcc gcctcggagt attttccatc
     7201 gaggtagatt aaagacatgc tcacccgagt tttatactct cctgcttgag atccttacta
     7261 cagtatgaaa ttacagtgtc gcgagttaga ctatgtaagc agaattttaa tcatttttaa
     7321 agagcccagt acttcatatc catttctccc gctccttctg cagccttatc aaaaggtatt
     7381 ttagaacact cattttagcc ccattttcat ttattatact ggcttatcca acccctagac
     7441 agagcattgg cattttccct ttcctgatct tagaagtctg atgactcatg aaaccagaca
     7501 gattagttac atacaccaca aatcgaggct gtagctgggg cctcaacact gcagttcttt
     7561 tataactcct tagtacactt tttgttgatc ctttgccttg atccttaatt ttcagtgtct
     7621 atcacctctc ccgtcaggtg gtgttccaca tttgggccta ttctcagtcc agggagtttt
     7681 acaacaatag atgtattgag aatccaacct aaagcttaac tttccactcc catgaatgcc
     7741 tctctccttt ttctccattt ataaactgaa gggcgaattc tgcagatatc catcacactg
     7801 gcggccgctc gatcgagcat gcatctagag ggcccaattc gccctatagt gagtcgtatt
     7861 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     7921 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     7981 ccgatcgccc ttcccaacag ttgcgcagcc tatacgtacg gcagtttaag gtttacacct
     8041 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
     8101 cggggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc
     8161 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
     8221 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
     8281 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggcatgagat
     8341 tatcaaaaag gatcttcacc tagatccttt tcacgtagaa agccagtccg cagaaacggt
     8401 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     8461 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     8521 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     8581 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagct
     8641 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     8701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     8761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     8821 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     8881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     8941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     9001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     9061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     9121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     9181 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     9241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     9301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     9361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     9421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt attaacgctt
     9481 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatca
     9541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     9601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atagcacgtg
     9661 aggagggcca ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     9721 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     9781 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     9841 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     9901 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     9961 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
    10021 cacttcgtgg ccgaggagca ggactgacac gtgctaaaac ttcattttta atttaaaagg
    10081 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    10141 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    10201 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    10261 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    10321 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    10381 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    10441 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    10501 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    10561 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    10621 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    10681 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    10741 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    10801 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    10861 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    10921 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    10981 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    11041 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    11101 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    11161 ggaaacagct atgaccatga ttacgccaag ctatttaggt gacactatag aatactcaag
    11221 ctatgcatca agcttggtac
//
