LOCUS       sequence-429277-       11280 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express a Lysophagy reporter in mammalian cells (untested).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11280)
  TITLE     Harper Lab plasmids
REFERENCE   2  (bases 1 to 11280)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pHAGE pGK LysoFlux (HaloTag7-mGFP-Galectin3)
FEATURES             Location/Qualifiers
     source          1..11280
                     /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(5299..6009,6010..6012))
                     /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(5341..5362)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5690..5709
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5949..5970
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(6034..6921)
                     /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      6918..6927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(6930..6954)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6961..7460)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7009..7028)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7402..7425
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    7556..7673
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7971..8015)
                     /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    8200..8433
                     /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    8930..9055
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             9102..9282
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(9300..9498)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(9325..9345)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        9500..9879
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     10051..10070
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        10145..10249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10250..10318,10319..11110)
                     /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(10468..10487)
                     /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 gttcattata tcatggtata tgaagcactg gtgaggtcta tgtcaccaga
     4561 aattcccagt ttgctgattt cattgagttt tttaacccga tgattgtact gcaacaagtg
     4621 agcatcattc actgcaacct tgaagtggtc aggttcaacc agtacttgta ttttgaatgg
     4681 tttcccactt tcaaatggga aaaccgactg tctttcttcc cttccccagt tattatccag
     4741 ctttgtattg caaacaatga ctctcctgtt gttctcattg aagcgtgggt taaagtggaa
     4801 ggcaacatca ttccctcttt ggaaatctaa agcaattctg tttgcattgg gcttcaccgt
     4861 gcccagaatt gttatcagca tgcgaggcac cactccccca ggcaaaggca ggttataagg
     4921 cacaatcagt ggcccagcag gggcgccata ggggccagtg gcagggtagg ctccgggggc
     4981 acttggctgt ccagaagatg ggtaggcccc agggccgctg ggtggccctg ggtagactcc
     5041 aggtgcaggt gctccgggat aagctccagg tgctccatgg taggcgcctg gaggtgcctg
     5101 tccaggataa gcccctgggg gtgcctgccc ggggtaggcc ccaggatagg aagcccctgg
     5161 gtagccccct gccccagcag gctggttccc ccatgcgcca ggccatcctt gagggtttgg
     5221 gtttccagac ccagataacg catcatggag cgaaaaattg tctgccatac ccgaattccc
     5281 agatcccgag cctgaaccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     5341 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagtttgga
     5401 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     5461 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     5521 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     5581 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     5641 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     5701 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     5761 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     5821 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     5881 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     5941 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     6001 gcccttgctc accccggatc ccgagcctga accaccggaa atctccagag tagacagcca
     6061 gcgcgcgatc tcgctgccga tcaggtccgg gttgtcttct tgcagcagat tcagacctgg
     6121 gccgatgtcc acagccttgc agttaggcag gcttttggcc aggcgagcgg cttcggccgg
     6181 tgggatcaga acgcctgggg tgccccagaa cagcagcttc gggacagggg actggtgcag
     6241 ccagtccatg tattcttcga ccagcgcgac gatgttcgct ggctcaccgg cgattggcag
     6301 ctcgtttggg aagcgccaca gtggctcgcg gtcaacagga ttcaggaacg gctcgcggta
     6361 atggtccatc tcgacttcag tcagcgggcg gacgacaccc atcggcagcg taccctcgat
     6421 aaaaacgttc tgatcgatga tcagcttgcg gccgacgtcg gtggtgcgga aggcctggaa
     6481 ggtctcgcgg gcaaattctg gccattcgtc ccaggtcggg atagggcgga tgaactccat
     6541 aaatgcaata cctttgacgc gctctggatt gcgcttggcc cagtggaaac ccagagcgga
     6601 gccccagtcg tgaatgacca ggacgacctc ttccagaccc agggcttcga tgaaggcatc
     6661 catgaagcgg acgtggtcgt cgaagaaata acccaggtct ggtttgtcgg atttgcccat
     6721 accgatcagg tctggagcaa tgcagcgatg ggtcggtgca acatgcggga tgatgttgcg
     6781 ccacacgtag gaggaggtcg ggttaccgtg caggaacagc acaggggtgc catcgcgcgg
     6841 accaacatcg acgtagtgca tgcgctcgcc caggacttcc acataatggg ggtcgaatgg
     6901 aaagccagta ccgatttcca tggtggcgga gcctgctttt ttgtacaaac ttgtttcgaa
     6961 cgaaaggccc ggagatgagg aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt
     7021 gcagaatgcc gggcctccgg aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct
     7081 gagcccgccc ccggacccac cccttcccag cctctgagcc cagaaagcga aggagcaaag
     7141 ctgctattgg ccgctgcccc aaaggcctac ccgcttccat tgctcagcgg tgctgtccat
     7201 ctgcacgaga ctagtgagac gtgctacttc catttgtcac gtcctgcacg acgcgagctg
     7261 cggggcgggg gggaacttcc tgactagggg aggagtagaa ggtggcgcga aggggccacc
     7321 aaagaacgga gccggttggc gcctaccggt ggatgtggaa tgtgtgcgag gccagaggcc
     7381 acttgtgtag cgccaagtgc ccagcggggc tgctaaagcg catgctccag actgccttgg
     7441 gaaaagcgcc tcccctaccc atagggataa cagggtaata taggcggccg cgtagatggc
     7501 gcgccggtac cttaattaac caaactggat ctctgctgtc cctgtaataa acccgaaaat
     7561 tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt
     7621 ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga
     7681 tgaatactgc catttgtctg cagaattggc gcacgcagtg ccgatccgtt cactaatcga
     7741 atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt
     7801 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     7861 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     7921 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     7981 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     8041 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     8101 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     8161 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     8221 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     8281 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     8341 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     8401 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     8461 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     8521 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
     8581 agcggccggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc tctatcttgt
     8641 ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag agggttgcta
     8701 ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt tgtagctgtc
     8761 ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact gcgaatcgtt
     8821 ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt ccccctggcc
     8881 ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg acgctctcgc
     8941 acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc accagtcgcc
     9001 gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga gagagctcct
     9061 ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga ttttccacac
     9121 tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac
     9181 tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc
     9241 agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg caaaacgcgc
     9301 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     9361 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     9421 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     9481 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     9541 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     9601 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     9661 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     9721 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     9781 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     9841 taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta
     9901 catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca
     9961 agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac
    10021 aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
    10081 accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa
    10141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
    10201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
    10261 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
    10321 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
    10381 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
    10441 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
    10501 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
    10561 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
    10621 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
    10681 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
    10741 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
    10801 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
    10861 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
    10921 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
    10981 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
    11041 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
    11101 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
    11161 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
    11221 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
