LOCUS       sequence-419318-       10887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express an autophagy (LC3) reporter in mammalian cells (untested).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10887)
  TITLE     Harper Lab plasmids
REFERENCE   2  (bases 1 to 10887)
  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: pHAGE pGK LC3Flux (HaloTag7-mGFP-LC3-STOP)
FEATURES             Location/Qualifiers
     source          1..10887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..1665)
                     /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"
     promoter        complement(1684..1731)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1779..2108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1793..1928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1847..1866)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2093..2113
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2122..2550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2444..2463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2596..2820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2809..2826
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             2852..3085
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3157..3745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3263..3274
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3672..3692
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3746..4144)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4141..4150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4169..4380)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    4488..4512
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4906..5616,5617..5619))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4948..4969)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5297..5316
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5556..5577
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5641..6528)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     regulatory      6525..6534
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(6537..6561)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6568..7067)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6616..6635)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7009..7032
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    7163..7280
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7578..7622)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7807..8040
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8537..8662
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             8709..8889
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8907..9105)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8932..8952)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        9107..9486
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9658..9677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9752..9856
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9857..9925,9926..10717)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10075..10094)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca cgaagtgcac
     1321 gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct cgccgatctc
     1381 ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg accactcggc
     1441 gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg gcaccacctg
     1501 gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc
     1561 ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg ctccggcgac
     1621 gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt agttcctcac
     1681 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     1741 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     1801 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     1861 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1921 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1981 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2041 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2101 tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta
     2161 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2221 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2281 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2341 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2401 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2461 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2521 ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc
     2581 tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
     2641 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     2701 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     2761 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     2821 tcgaggctga tcagcgggtt taaacgggcc ctgctagaga ttttccacac tgactaaaag
     2881 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     2941 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     3001 caggctcaga tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt
     3061 cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct
     3121 acagctgcct tgtaagtcat tggtcttaaa gtcgacgcgg ggaggcggcc caaagggaga
     3181 tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc
     3241 cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc
     3301 gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca
     3361 acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
     3421 cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct
     3481 gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca
     3541 cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat
     3601 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
     3661 cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac
     3721 aaattttgta atccagaggt tgattttagc cctcccacac ataaccagag ggcagcaatt
     3781 cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag
     3841 gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct
     3901 catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca
     3961 gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga
     4021 tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc
     4081 tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg
     4141 ccatggtggc agcgctctag aaccggtcct gtgttctggc ggcaaacccg ttgcgaaaaa
     4201 gaacgttcac ggcgactact gcacttatat acggttctcc cccaccctcg ggaaaaaggc
     4261 ggagccagta cacgacatca ctttcccagt ttaccccgcg ccaccttctc taggcaccgg
     4321 atcaattgcc gacccctccc cccaacttct cggggactgt gggcgatgtg cgctctgccc
     4381 actgacgggc accggagcca attcccactc ctttcaagac ctagctagtc ctgcaggttc
     4441 gctaccttag gaccgttata gttacgctcg aggtttaaac tcattaaacc actttgtaca
     4501 agaaagctgg gttcattacc cgaacgtctc ctgggaggca tagaccatgt acaggaagcc
     4561 gtcttcatct ctctcgctct cgtacacttc agagatgggt gtggacacac tcaccatgct
     4621 gtgcccattc accaggagga agaaggcttg gttagcattg agctgcaggc gccttctaat
     4681 tatcttgatg agttcgctca tattcacgtg atcaggtaca aggaacttgg tcttgtccag
     4741 gacgggcagc tgcttctcac ccttgtatcg ctctataatc actgggatct tggtggggtg
     4801 ctgctcccgg atgagccgga catcttccac tctttgttcg aagctccggc gctgtttgaa
     4861 ggtcttctcg gacggacccg aattcccaga tcccgagcct gaacccttgt acagctcgtc
     4921 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     4981 ctcgttgggg tctttgctca gtttggactg ggtgctcagg tagtggttgt cgggcagcag
     5041 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     5101 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     5161 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     5221 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     5281 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     5341 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     5401 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     5461 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     5521 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     5581 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc ccggatcccg agcctgaacc
     5641 accggaaatc tccagagtag acagccagcg cgcgatctcg ctgccgatca ggtccgggtt
     5701 gtcttcttgc agcagattca gacctgggcc gatgtccaca gccttgcagt taggcaggct
     5761 tttggccagg cgagcggctt cggccggtgg gatcagaacg cctggggtgc cccagaacag
     5821 cagcttcggg acaggggact ggtgcagcca gtccatgtat tcttcgacca gcgcgacgat
     5881 gttcgctggc tcaccggcga ttggcagctc gtttgggaag cgccacagtg gctcgcggtc
     5941 aacaggattc aggaacggct cgcggtaatg gtccatctcg acttcagtca gcgggcggac
     6001 gacacccatc ggcagcgtac cctcgataaa aacgttctga tcgatgatca gcttgcggcc
     6061 gacgtcggtg gtgcggaagg cctggaaggt ctcgcgggca aattctggcc attcgtccca
     6121 ggtcgggata gggcggatga actccataaa tgcaatacct ttgacgcgct ctggattgcg
     6181 cttggcccag tggaaaccca gagcggagcc ccagtcgtga atgaccagga cgacctcttc
     6241 cagacccagg gcttcgatga aggcatccat gaagcggacg tggtcgtcga agaaataacc
     6301 caggtctggt ttgtcggatt tgcccatacc gatcaggtct ggagcaatgc agcgatgggt
     6361 cggtgcaaca tgcgggatga tgttgcgcca cacgtaggag gaggtcgggt taccgtgcag
     6421 gaacagcaca ggggtgccat cgcgcggacc aacatcgacg tagtgcatgc gctcgcccag
     6481 gacttccaca taatgggggt cgaatggaaa gccagtaccg atttccatgg tggcggagcc
     6541 tgcttttttg tacaaacttg tttcgaacga aaggcccgga gatgaggaag aggagaacag
     6601 cgcggcagac gtgcgctttt gaagcgtgca gaatgccggg cctccggagg accttcgggc
     6661 gcccgccccg cccctgagcc cgcccctgag cccgcccccg gacccacccc ttcccagcct
     6721 ctgagcccag aaagcgaagg agcaaagctg ctattggccg ctgccccaaa ggcctacccg
     6781 cttccattgc tcagcggtgc tgtccatctg cacgagacta gtgagacgtg ctacttccat
     6841 ttgtcacgtc ctgcacgacg cgagctgcgg ggcggggggg aacttcctga ctaggggagg
     6901 agtagaaggt ggcgcgaagg ggccaccaaa gaacggagcc ggttggcgcc taccggtgga
     6961 tgtggaatgt gtgcgaggcc agaggccact tgtgtagcgc caagtgccca gcggggctgc
     7021 taaagcgcat gctccagact gccttgggaa aagcgcctcc cctacccata gggataacag
     7081 ggtaatatag gcggccgcgt agatggcgcg ccggtacctt aattaaccaa actggatctc
     7141 tgctgtccct gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt
     7201 tagtttgtat gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa
     7261 tccccccttt tcttttaaaa ttgtggatga atactgccat ttgtctgcag aattggcgca
     7321 cgcagtgccg atccgttcac taatcgaatg gatctgtctc tgtctctctc tccaccttct
     7381 tcttctattc cttcgggcct gtcgggtccc ctcggggttg ggaggtgggt ctgaaacgat
     7441 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     7501 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     7561 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     7621 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     7681 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     7741 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     7801 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     7861 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     7921 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     7981 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     8041 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     8101 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     8161 atatctcctc ctccaggtct gaagatcagc ggccggccgc ttgctgtgcg gtggtcttac
     8221 ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc
     8281 tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc
     8341 ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta
     8401 acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta
     8461 atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc
     8521 tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca
     8581 aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa
     8641 gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg
     8701 ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc
     8761 agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg
     8821 cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca
     8881 gagagaccca gtacaggcaa aacgcgctgc ttatatagac ctcccaccgt acacgcctac
     8941 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     9001 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     9061 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     9121 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     9181 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     9241 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     9301 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     9361 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     9421 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     9481 aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa
     9541 gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg
     9601 cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg
     9661 catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc
     9721 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     9781 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     9841 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     9901 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     9961 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
    10021 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
    10081 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
    10141 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
    10201 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
    10261 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
    10321 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
    10381 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
    10441 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
    10501 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
    10561 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
    10621 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
    10681 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
    10741 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
    10801 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
    10861 ccccgtagaa aagatcaaag gatcttc
//
