LOCUS       sequence-356653-       18069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  targeting vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18069)
  TITLE     Daigle Lab TIGRE3.0 plasmids 2022
REFERENCE   2  (bases 1 to 18069)
  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"
COMMENT     Sequence Label: AiP1069-Ai210 (TITL-GC7f-ICF-IRES-tTA2)
FEATURES             Location/Qualifiers
     source          1..18069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..79
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             98..901
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(161..180)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     771..790
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     930..952
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     944..961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     945..961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1105..1154
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     promoter        4304..4618
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4312..4330
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4348..4366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4383..4401
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4419..4437
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4455..4473
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4490..4508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4526..4544
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4591..4615
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(4628..4661)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4738..4786
                     /note="synthetic polyadenylation signal"
     polyA_signal    4795..5271
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    5283..5744
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(5771..5804)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5815..5824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5821..5829,5830..5847,5848..5850,5851..5883,5884..5886,
                     5887..5910,5911..5937,5938..5994,5995..6000,6001..6270,
                     6271..6288,6289..6723,6724..6729,6730..7173)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      5917..5926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     6207..6228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7198..7786
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(7669..7680)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7817..8024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        10548..10851
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10853..11130
                     /label=chicken beta-actin promoter
     intron          11131..12147
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     12071..12093
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     12155..12174
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    12235..12268
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    12314..12362
                     /note="synthetic polyadenylation signal"
     polyA_signal    12371..12847
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(12882..12901)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    12926..12974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    13134..13167
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    13190..13763
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    13213..13765
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(13357..13374)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     13584..13603
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(13773..14390,14391..14396,14397..14432,14433..14435,
                     14436..14471,14472..14474,14475..14510,14511..14519)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    14560..15148
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(15031..15042)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    15179..15386
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    15400..15433
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        15460..15959
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(15495..15518)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     15892..15911
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    16651..16700
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     rep_origin      complement(17013..17601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(17093..17112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgttttgg cggtttaaac attaagcacg tgttgacaat taatcatcgg catagtatat
       61 cggcatagta taatacgaca aggtgaggaa ctaaaccatg ggatcggcca ttgaacaaga
      121 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
      181 acaacagacg atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
      241 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
      301 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
      361 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
      421 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
      481 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
      541 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
      601 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
      661 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
      721 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
      781 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
      841 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
      901 aagtgttaac tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      961 tgaattcgag ctcggtaccg aaaaaccgct cagcggcgcc cgcacccaaa cgaaaacacc
     1021 cccggcccaa gcggaccggg ggtgtcgcgt gaaatggatc agtgagcgtg gccgtggtgg
     1081 tggccgtggc cgtgctcgtc ctcgtcggcc ggcttgtcga cgacggcggt ctccgtcgtc
     1141 aggatcatcc gggccaccga ggcggcgttg agaacagccg agcgggtcac cttggccgga
     1201 tcgacgacgc cggcggagac cagatcaccg aactccagcg tggcggcgtt gaagccctgc
     1261 cccttgggaa gttcggagat ttcggccggc caatcgattt taccggggtg gtgcccatcc
     1321 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1381 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1441 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1501 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1561 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1621 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1681 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1741 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggtt
     1801 aattaagaat tcgagctcgc tagagcccca tcctcactga ctccgtcctg gagttggatg
     1861 agagataatg gccttacgtt gtgccagggg agggtcgggc tggatttagc aagatttacc
     1921 ttctccaaag agcggtgctg cagtggcaca gctgcccacg gaggtggggg ggtcaccgtc
     1981 cctggaggtg atgaagaact gtggggatgt ggcactgagg gacatggcca gtgggcacgg
     2041 tgggtgggtt ggggttggtc ttggggatct tggagggctt ttccagcctt catgatttga
     2101 cgattgtatg aacatctaca tggcaattct ccagctgcct gtcccagtcc tactgaccca
     2161 gctgtatctc tccaggcaag ctcttccacc ccttctgctt gcatccagac accatcaaac
     2221 atgcaggctc agacacaggg accagcagtg tctgtggcct ttttgtgctc ctctccatgc
     2281 tgggttttaa cttgctcttt gtccttctat cctatcttct tatccttaag gctgttctga
     2341 acgctgtgac ttggagagtg tcccagagcc ctcaacacct gcatgtccca cgtccatgct
     2401 gtcctgcact tccttatccc caagatctgc ctctccgtga tgcactgaat tggcaaacat
     2461 gtgtcacccc agaccaacaa tgtcacagca aactccccct tgataggaca agggggaatg
     2521 gctttacact gagacagggg aggtttgggt tggatatgag gaggcagttt ttcccccaga
     2581 gggtggtgac gcactgaaca ggttgcccaa ggaggctgtg gatgccccat ccctgcaggc
     2641 attcaaggcc aggctggatg tggctctggg cagcctgggc tgctggttga tgaccctgca
     2701 catagcaggg ggttggatct ggatgagcac tgtgctcctt tgcaacccag gccgttctat
     2761 gattctgtca ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt
     2821 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
     2881 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
     2941 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
     3001 cctgggggct ttgggggggg gctgtccctc tagcgagctc ggtacccaga gccccatcct
     3061 cactgactcc gtcctggagt tggatgagag ataatggcct tacgttgtgc caggggaggg
     3121 tcgggctgga tttagcaaga tttaccttct ccaaagagcg gtgctgcagt ggcacagctg
     3181 cccacggagg tgggggggtc accgtccctg gaggtgatga agaactgtgg ggatgtggca
     3241 ctgagggaca tggccagtgg gcacggtggg tgggttgggg ttggtcttgg ggatcttgga
     3301 gggcttttcc agccttcatg atttgacgat tgtatgaaca tctacatggc aattctccag
     3361 ctgcctgtcc cagtcctact gacccagctg tatctctcca ggcaagctct tccacccctt
     3421 ctgcttgcat ccagacacca tcaaacatgc aggctcagac acagggacca gcagtgtctg
     3481 tggccttttt gtgctcctct ccatgctggg ttttaacttg ctctttgtcc ttctatccta
     3541 tcttcttatc cttaaggctg ttctgaacgc tgtgacttgg agagtgtccc agagccctca
     3601 acacctgcat gtcccacgtc catgctgtcc tgcacttcct tatccccaag atctgcctct
     3661 ccgtgatgca ctgaattggc aaacatgtgt caccccagac caacaatgtc acagcaaact
     3721 cccccttgat aggacaaggg ggaatggctt tacactgaga caggggaggt ttgggttgga
     3781 tatgaggagg cagtttttcc cccagagggt ggtgacgcac tgaacaggtt gcccaaggag
     3841 gctgtggatg ccccatccct gcaggcattc aaggccaggc tggatgtggc tctgggcagc
     3901 ctgggctgct ggttgatgac cctgcacata gcagggggtt ggatctggat gagcactgtg
     3961 ctcctttgca acccaggccg ttctatgatt ctgtcattct aaatctctct ttcagcctaa
     4021 agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca
     4081 gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct
     4141 cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc
     4201 tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg tccctctagg
     4261 ggtacccggg gatccaccgg tatcgattac ctatcacaaa ttcgagttta ctccctatca
     4321 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
     4381 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     4441 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
     4501 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
     4561 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgccta
     4621 tttaaatata acttcgtata gcatacatta tacgaagtta ttcgcgatga ataaatgaaa
     4681 gcttgcagat cccctcagca ccatggctag cggcagcctc ggagtttgaa tagatagaat
     4741 aaaatatctt tattttcatt ccatctgtgt gttggttttt tgtgtgagat ctacgggtgg
     4801 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4861 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4921 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4981 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     5041 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc aggctcccga gttgttggga
     5101 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     5161 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     5221 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgcctcag
     5281 cgctgtaagt ctgcagaaat tgatgatcta ttaaacaata aagatgtcca ctaaaatgga
     5341 agtttttcct gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg
     5401 aaggattgga gctacggggg tgggggtggg gtgggattag ataaatgcct gctctttact
     5461 gaaggctctt tactattgct ttatgataat gtttcatagt tggatatcat aatttaaaca
     5521 agcaaaacca aattaagggc cagctcattc ctcccactca tgatctatag atctatagat
     5581 ctctcgtggg atcattgttt ttctcttgat tcccactttg tggttctaag tactgtggtt
     5641 tccaaatgtg tcagtttcat agcctgaaga acgagatcag cagcctctgt tccacataca
     5701 cttcattctc agtattgttt tgccaagttc taattccatc agaaagcttt tacttaactg
     5761 aatagctaac ataacttcgt atagcataca ttatacgaag ttatcacgcg tgccgccacc
     5821 atgggttctc atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg
     5881 ggtcgggatc tgtacgacga tgacgataag gatctcgcca ccatggtcga ctcatcacgt
     5941 cgtaagtgga ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag
     6001 aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt caagatccgc
     6061 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
     6121 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg
     6181 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     6241 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
     6301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     6361 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     6421 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     6481 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     6541 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     6601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     6661 gagcttaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     6721 aacctgccgg accaactgac tgaagagcag atcgcagaat ttaaagagct gttctcccta
     6781 tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct
     6841 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac
     6901 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac
     6961 acagacagtg aagaagaaat tagagaagcg ttccgtgtgt ttgataagga tggcaatggc
     7021 tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     7081 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac
     7141 gaagagtttg tacaaatgat gacagcgaag tgaacgcgtt acgtaaagct tgtcgataat
     7201 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     7261 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     7321 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     7381 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     7441 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7501 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     7561 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     7621 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     7681 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     7741 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg acaccgagcg
     7801 ctgctcgagg cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     7861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     7921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     7981 agcaaggggg aggattggga agacaatagc aggcatgctg gggacacgtg ggatcctcta
     8041 gagccccatc ctcactgact ccgtcctgga gttggatgag agataatggc cttacgttgt
     8101 gccaggggag ggtcgggctg gatttagcaa gatttacctt ctccaaagag cggtgctgca
     8161 gtggcacagc tgcccacgga ggtggggggg tcaccgtccc tggaggtgat gaagaactgt
     8221 ggggatgtgg cactgaggga catggccagt gggcacggtg ggtgggttgg ggttggtctt
     8281 ggggatcttg gagggctttt ccagccttca tgatttgacg attgtatgaa catctacatg
     8341 gcaattctcc agctgcctgt cccagtccta ctgacccagc tgtatctctc caggcaagct
     8401 cttccacccc ttctgcttgc atccagacac catcaaacat gcaggctcag acacagggac
     8461 cagcagtgtc tgtggccttt ttgtgctcct ctccatgctg ggttttaact tgctctttgt
     8521 ccttctatcc tatcttctta tccttaaggc tgttctgaac gctgtgactt ggagagtgtc
     8581 ccagagccct caacacctgc atgtcccacg tccatgctgt cctgcacttc cttatcccca
     8641 agatctgcct ctccgtgatg cactgaattg gcaaacatgt gtcaccccag accaacaatg
     8701 tcacagcaaa ctcccccttg ataggacaag ggggaatggc tttacactga gacaggggag
     8761 gtttgggttg gatatgagga ggcagttttt cccccagagg gtggtgacgc actgaacagg
     8821 ttgcccaagg aggctgtgga tgccccatcc ctgcaggcat tcaaggccag gctggatgtg
     8881 gctctgggca gcctgggctg ctggttgatg accctgcaca tagcaggggg ttggatctgg
     8941 atgagcactg tgctcctttg caacccaggc cgttctatga ttctgtcatt ctaaatctct
     9001 ctttcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc
     9061 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc
     9121 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc
     9181 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc
     9241 tgtccctcta gagccccatc ctcactgact ccgtcctgga gttggatgag agataatggc
     9301 cttacgttgt gccaggggag ggtcgggctg gatttagcaa gatttacctt ctccaaagag
     9361 cggtgctgca gtggcacagc tgcccacgga ggtggggggg tcaccgtccc tggaggtgat
     9421 gaagaactgt ggggatgtgg cactgaggga catggccagt gggcacggtg ggtgggttgg
     9481 ggttggtctt ggggatcttg gagggctttt ccagccttca tgatttgacg attgtatgaa
     9541 catctacatg gcaattctcc agctgcctgt cccagtccta ctgacccagc tgtatctctc
     9601 caggcaagct cttccacccc ttctgcttgc atccagacac catcaaacat gcaggctcag
     9661 acacagggac cagcagtgtc tgtggccttt ttgtgctcct ctccatgctg ggttttaact
     9721 tgctctttgt ccttctatcc tatcttctta tccttaaggc tgttctgaac gctgtgactt
     9781 ggagagtgtc ccagagccct caacacctgc atgtcccacg tccatgctgt cctgcacttc
     9841 cttatcccca agatctgcct ctccgtgatg cactgaattg gcaaacatgt gtcaccccag
     9901 accaacaatg tcacagcaaa ctcccccttg ataggacaag ggggaatggc tttacactga
     9961 gacaggggag gtttgggttg gatatgagga ggcagttttt cccccagagg gtggtgacgc
    10021 actgaacagg ttgcccaagg aggctgtgga tgccccatcc ctgcaggcat tcaaggccag
    10081 gctggatgtg gctctgggca gcctgggctg ctggttgatg accctgcaca tagcaggggg
    10141 ttggatctgg atgagcactg tgctcctttg caacccaggc cgttctatga ttctgtcatt
    10201 ctaaatctct ctttcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc
    10261 aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg
    10321 ctgtccccgc acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc
    10381 cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt
    10441 gggggggggc tgtccctcta gagtcgacgg cgcgccctgc aggaatctag ttattaatag
    10501 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
    10561 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
    10621 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
    10681 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
    10741 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
    10801 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
    10861 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
    10921 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
    10981 ccaggsgggg cggggsgggg sgrggggsgg ggsggggcga ggcggagagg tgcggcggca
    11041 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
    11101 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
    11161 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
    11221 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
    11281 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
    11341 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
    11401 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
    11461 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc
    11521 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc
    11581 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg
    11641 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg
    11701 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg
    11761 gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat
    11821 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
    11881 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
    11941 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
    12001 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
    12061 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
    12121 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
    12181 catcattttg gcaaagaatt gatttgatac cgcgggcccc gcgtggacgg taccgaagtt
    12241 cctattctct agaaagtata ggaacttccc tcagcaccat ggctagcggc agcctcggag
    12301 tttgaataga tagaataaaa tatctttatt ttcattccat ctgtgtgttg gttttttgtg
    12361 tgagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
    12421 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
    12481 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
    12541 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
    12601 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcaggc
    12661 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
    12721 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
    12781 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
    12841 tgtccttctg cctcagctaa ttgagtaggg gggaggctaa ctgaaacacg gaaggagaca
    12901 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
    12961 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
    13021 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
    13081 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc gtggaagttc
    13141 ctattctcta gaaagtatag gaacttcacg cgcgcgtcca ttaattccgc ccctctccct
    13201 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
    13261 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
    13321 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
    13381 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
    13441 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
    13501 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
    13561 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
    13621 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta
    13681 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
    13741 tcctttgaaa aacacgatga taataattca ccatgtctag actggacaag agcaaagtca
    13801 taaactctgc tctggaatta ctcaatgaag tcggtatcga aggcctgacg acaaggaaac
    13861 tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg
    13921 ccctgctcga tgccctggca atcgagatgc tggacaggca tcatacccac ttctgccccc
    13981 tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc
    14041 tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt
    14101 acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg
    14161 cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggatcagg
    14221 agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc
    14281 tgagacaagc aattgagctg ttcgaccatc agggagccga acctgccttc cttttcggcc
    14341 tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg
    14401 acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc
    14461 ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaac
    14521 taaaccaggt aaatggcgcg ttacgtaaag cttgtcgata atcaacctct ggattacaaa
    14581 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
    14641 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
    14701 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
    14761 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
    14821 tgtcaactcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
    14881 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
    14941 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
    15001 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
    15061 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
    15121 cggatctccc tttgggccgc ctccccgcat cgacaccgag cgctgctcga ggcctcgact
    15181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
    15241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
    15301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
    15361 gaagacaata gcaggcatgc tggggacacg tgggatccgg tgccagggcg tgcccttggg
    15421 ctccccgggc gcgtttgggc ggccgcatcg aattctaccg ggtaggggag gcgcttttcc
    15481 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
    15541 gcctctggcc tcgcacacat tccacatcct ccggtaggcg ccaaccggct ccgttctttg
    15601 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
    15661 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
    15721 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
    15781 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
    15841 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
    15901 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
    15961 cctgcaatcg ccgctagccg ccaccatgaa aaagcctgaa ctcaccgcga cgtctgtcga
    16021 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
    16081 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
    16141 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
    16201 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
    16261 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
    16321 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
    16381 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
    16441 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgt
    16501 aagtgctagc ccctaggcct gaatcgaaaa accgctcagc ggcgcccgca cccaaacgaa
    16561 aacacccccg gcccaagcgg accgggggtg tcgcgtgaaa tggatcagtg agcgtggccg
    16621 tggtggtggc cgtggccgtg ctcgtcctcg tcggccggct tgtcgacgac ggcggtctcc
    16681 gtcgtcagga tcatccgggc caccgaggcg gcgttgagaa cagccgagcg ggtcaccttg
    16741 gccggatcga cgacgccggc ggagaccaga tcaccgaact ccagcgtggc ggcgttgaag
    16801 ccctgcccct tgggaagttc ggagagagct cgtcgactgc agaggcctgc atgcaagctt
    16861 ggcgtaatca tggtcatagc tgtttcctta tcagctcact caaaggcggt aatacggtta
    16921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    16981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    17041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    17101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    17161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    17221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    17281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    17341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    17401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    17461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    17521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    17581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    17641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    17701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    17761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    17821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    17881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    17941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    18001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    18061 agtttgcgc
//
