LOCUS       sequence-12685-e       12926 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12926)
  AUTHORS   Hsiao EC, Nguyen TD, Ng JK, Scott MJ, Chang WC, Zahed H, Conklin BR
  TITLE     Constitutive Gs activation using a single-construct 
            tetracycline-inducible expression system in embryonic stem cells and
            mice.
  JOURNAL   Stem Cell Res Ther. 2011 Mar 4. 2(2):11.
  PUBMED    21375737
REFERENCE   2  (bases 1 to 12926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cell 
            Res Ther. 2011 Mar 4. 2(2):11."
COMMENT     Sequence Label: Exp-pcDNA3.2delCMV(EF1-alpha-tTA/TetO-mCh-Rs1)
FEATURES             Location/Qualifiers
     source          1..12926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..81
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(121..140)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    165..213
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      415..843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(502..521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     712..733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(852..872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        857..1186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1037..1172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1099..1118
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1253..2047
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1307..1326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1917..1936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2223..2344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2260..2279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2314..2333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2393..2409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2393..2409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2406..2428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2417..2433
                     /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)."
     promoter        complement(join(2441..2447,2448..2465,2466..2471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2486..2507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2795..3383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3554..4345,4346..4414))
                     /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     4177..4196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4415..4519)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4594..4613)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4844..4863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4844..4862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4892..4916
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4949..4982)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5673..5884
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5897..6165
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             join(6221..6841,6842..7228)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(6294..6313)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     intron          7265..7837
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(7969..7988)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8034..8089
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8088..8107)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    complement(8928..8948)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    8975..9023
                     /note="synthetic polyadenylation signal"
     CDS             complement(10387..10443)
                     /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             complement(10468..11175)
                     /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(10743..10762)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     11012..11030
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      11172..11181
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          11254..11826
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(11939..11963)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(11960..11998)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    12031..12049
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12073..12091
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12115..12133
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12157..12175
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(12861..12894)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(join(12916..12926,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttgatct agagggcccg cggttcgaag gtaagcctat ccctaaccct
       61 ctcctcggtc tcgattctac gcgtaccggt tagtaatgag tttaaacggg ggaggctaac
      121 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
      181 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
      241 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
      301 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
      361 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
      421 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      481 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      541 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
      601 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      661 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      721 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      781 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      841 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
      901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
      961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     1021 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1081 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     1141 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     1201 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1261 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1321 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1381 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1441 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1501 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1561 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1621 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     1681 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1741 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     1801 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1861 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     1921 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1981 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2041 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     2101 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     2161 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     2221 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2281 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2341 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     2401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     3001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     4561 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     4621 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     4681 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     4741 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     4801 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     4861 gggagaccca agctggctag ttaagctatc aacaagtttg tacaaaaaag caggctggcg
     4921 ccggaaccaa ttcagtcgac gtttaaacat aacttcgtat agcatacatt atacgaagtt
     4981 atctcgagcc tcaggaaaca gggtaccagt gaaagtctgc aatgaattcg agttggcgcg
     5041 cctgtcattc taaatctctc tttcagccta aagctttttc cccgtatccc cccaggtgtc
     5101 tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc
     5161 gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga
     5221 gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct
     5281 gggggctttg ggagggggct gtccccgtga gctcccaggc gcgcctgtca ttctaaatct
     5341 ctctttcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca
     5401 gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc
     5461 gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc
     5521 tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg
     5581 gctgtccccg tgagctccca ggcgcgccaa ctcggtaccc ggggatccac atatgttaat
     5641 taagatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     5701 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     5761 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     5821 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     5881 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
     5941 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
     6001 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
     6061 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
     6121 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
     6181 gcctacaagg cctgcaggtc gacggatccg gggaattcat atgtctagat tagataaaag
     6241 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac
     6301 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa
     6361 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt
     6421 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag
     6481 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga
     6541 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact
     6601 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga
     6661 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc
     6721 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt
     6781 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc
     6841 cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga
     6901 tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct
     6961 ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga
     7021 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt
     7081 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc
     7141 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc
     7201 ccttggaatt gacgagtacg gtgggtaggg ggcgcgagga tcctctagag ctgagaactt
     7261 cagggtgagt ttggggaccc ttgattgttc tttctttttc gctattgtaa aattcatgtt
     7321 atatggaggg ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc ccttgtatca
     7381 ccatggaccc tcatgataat tttgtttctt tcactttcta ctctgttgac aaccattgtc
     7441 tcctcttatt ttcttttcat tttctgtaac tttttcgtta aactttagct tgcatttgta
     7501 acgaattttt aaattcactt ttgtttattt gtcagattgt aagtactttc tctaatcact
     7561 tttttttcaa ggcaatcagg gtatattata ttgtacttca gcacagtttt agagaacaat
     7621 tgttataatt aaatgataag gtagaatatt tctgcatata aattctggct ggcgtggaaa
     7681 tattcttatt ggtagaaaca actacaccct ggtcatcatc ctgcctttct ctttatggtt
     7741 acaatgatat acactgtttg agatgaggat aaaatactct gagtccaaac cgggcccctc
     7801 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttgt
     7861 tgtgctgtct catcattttg gcaaagaatt cactcctcag gtgcaggctg cctatcagaa
     7921 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     7981 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg ggtaataaag
     8041 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     8101 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     8161 atatgccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     8221 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     8281 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     8341 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     8401 ctcttatgga gatccctcga ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     8461 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     8521 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     8581 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     8641 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     8701 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     8761 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     8821 ttctcccttc gggaagcgtg gcgctttctc aattagctca cgctgtaggt atctcagttc
     8881 ggtgtaggtc gttcgctcca agctgggctg tgcgcggccg cagatctcaa ctttattata
     8941 catagttgct taagatctct gaatgcggcc gcttcacaca aaaaaccaac acacagatgt
     9001 aatgaaaata aagatatttt attgcggcca tcgtgatggc aggttgggcg tcgcttggtc
     9061 ggtcatttcg aaccccagag tcccgcctgc aggtccggaa ctagtctaag tgtcactggg
     9121 ctgagcagcc accaaaggag aagttgctgg cgggtgacac tgactctccc actggccacc
     9181 acactccact gcatccctta gtacatgtgt ggatccatta atggttgtgg ttgaacaagg
     9241 gacagtctgg cccagaatgg aaggtcttcg gtagcgctca tcatcacagc agaggatgat
     9301 gaggaaggca cgtctaaaag acttattcaa gaaggcgtag agaaaagggt tcaacccgga
     9361 attgatatag ccgagccaga ggaaagcagt ccacacctgc ccagggacag tgtagtctat
     9421 gaaaggatcc acaatattgg tgacaaagaa tggtgcccag cagaggcaga agcaacccat
     9481 gatgatgcat agggtcttgg ctgctttggt ctctgtcctc atgcgatgag tgctatgctg
     9541 gtctgccgac tgaggcctgc tctcggagga ggctcctgcc cgttgtaaca tctggatctg
     9601 atgggcatgc tccttagctg tgacatagat gcggtaatag gccagcacca tgaggagaaa
     9661 tgggatgtag aaggccacca cagagcaggt gatggcgtag ggcttgttga ccatgaagac
     9721 acagtacgta gagttagagt tctggttgaa cttcctcttt tctatcaaat caattatgcc
     9781 aatgttattc cagccttgca ttatagggag aaaagaaata aacgtgggga tgacccagca
     9841 gcctcccagc attaatgcga tgcgcagagg ggtcatcttg ttcctataga ccaaaggctg
     9901 gcagcagatg gcgtaatacc tatccagaga aatgcagcac aggtgaaaaa tcgatgccgt
     9961 tgtgagcagg acggccagag atgtccgaac aagacaaaac acctccccat aaatccagat
    10021 gtcttgaacc agctcaatgg caccaaaggg catcaccagc accgaaacca gcagatccgc
    10081 aaaagcaaga gatacaatga aataatttgt ttttattttc ctgagctgcc tgtcccagca
    10141 cacagccacc atcaccagca ggttccccaa gatggccatc aggataaccg tcgagagaaa
    10201 cgtgagcagc accaccttct ccactgaccc gaaaccctcc tcagaactca cattagcatc
    10261 aagtttgtcc atcctgcagc ccgggggatc cgcccgggcg cggccgctgg cgtcatcatc
    10321 gtccttgtag tcggcgaata ccaggcagaa gatgtagctc agggcgatga tcgtcttgat
    10381 accggtcggt cctgggtttt cttccacatc gcccgcctgt ttcagcaggc taaagttggt
    10441 cgcgccgctg ccctcgagaa accaggtctt gtacagctcg tccatgccgc cggtggagtg
    10501 gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
    10561 gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta
    10621 ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat
    10681 ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat
    10741 ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
    10801 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
    10861 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc
    10921 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag
    10981 gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt
    11041 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
    11101 gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc
    11161 gcccttgctc accatggtgg cggcgcctag gcagtcgacg aattctttgc caaaatgatg
    11221 agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga aggcatgaac
    11281 atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct caaacagtgt
    11341 atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt tgtttctacc
    11401 aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac cttatcattt
    11461 aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac cctgattgcc
    11521 ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa aagtgaattt
    11581 aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa atgaaaagaa
    11641 aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa attatcatga
    11701 gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga aaactttgcc
    11761 ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa gggtccccaa
    11821 actcaccctg aagttctcag gatccccggg taccgagctc gaattcgggg ccgcggaggc
    11881 tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga
    11941 tctgacggtt cactaaacga gctctgctta tataggcctc ccaccgtaca cgcctacctc
    12001 gacccgggta ccgagctcga ctttcacttt tctctatcac tgatagggag tggtaaactc
    12061 gactttcact tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta
    12121 tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt
    12181 aaactcgagt gaagacgaaa gggccttaat taacctcagg aaacagggta ccagtgaaag
    12241 tctgcaatga attcgagttg gcgcgcctgt cattctaaat ctctctttca gcctaaagct
    12301 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
    12361 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
    12421 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
    12481 gggggaggga cgtaattaca tccctggggg ctttgggagg gggctgtccc cgtgagctcc
    12541 caggcgcgcc tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc
    12601 caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca
    12661 ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc
    12721 ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt
    12781 acatccctgg gggctttggg ggggggctgt ccccgtgagc tcccaggcgc gccaactcgg
    12841 tacccgggga tccacatatg ataacttcgt atagcataca ttatacgaag ttatgtttaa
    12901 actcgagata tctagaccca gctttc
//
