LOCUS       sequence-429268-       10557 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express a ERphagy reporter in mammalian cells (untested).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10557)
  TITLE     Harper Lab plasmids
REFERENCE   2  (bases 1 to 10557)
  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 ERFlux (Bip-HaloTag7-mGFP-KDEL-STOP)
FEATURES             Location/Qualifiers
     source          1..10557
                     /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(4537..5247,5248..5250))
                     /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(4579..4600)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4928..4947
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5187..5208
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5257..6141)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     sig_peptide     complement(6145..6198)
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     protein_bind    complement(6207..6231)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6238..6737)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6286..6305)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6679..6702
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    6833..6950
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7248..7292)
                     /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    7477..7710
                     /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    8207..8332
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             8379..8559
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8577..8775)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8602..8622)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        8777..9156
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9328..9347
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9422..9526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9527..9595,9596..10387)
                     /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(9745..9764)
                     /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 gtttatcaga gctcatcttt tgaattcttg tacagctcgt ccatgccgag
     4561 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     4621 gtctttgctc agtttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     4681 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     4741 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     4801 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     4861 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     4921 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     4981 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     5041 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     5101 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     5161 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     5221 cccggtgaac agctcctcgc ccttgctcac ctcgagaccg gaaatctcca gagtagacag
     5281 ccagcgcgcg atctcgctgc cgatcaggtc cgggttgtct tcttgcagca gattcagacc
     5341 tgggccgatg tccacagcct tgcagttagg caggcttttg gccaggcgag cggcttcggc
     5401 cggtgggatc agaacgcctg gggtgcccca gaacagcagc ttcgggacag gggactggtg
     5461 cagccagtcc atgtattctt cgaccagcgc gacgatgttc gctggctcac cggcgattgg
     5521 cagctcgttt gggaagcgcc acagtggctc gcggtcaaca ggattcagga acggctcgcg
     5581 gtaatggtcc atctcgactt cagtcagcgg gcggacgaca cccatcggca gcgtaccctc
     5641 gataaaaacg ttctgatcga tgatcagctt gcggccgacg tcggtggtgc ggaaggcctg
     5701 gaaggtctcg cgggcaaatt ctggccattc gtcccaggtc gggatagggc ggatgaactc
     5761 cataaatgca atacctttga cgcgctctgg attgcgcttg gcccagtgga aacccagagc
     5821 ggagccccag tcgtgaatga ccaggacgac ctcttccaga cccagggctt cgatgaaggc
     5881 atccatgaag cggacgtggt cgtcgaagaa ataacccagg tctggtttgt cggatttgcc
     5941 cataccgatc aggtctggag caatgcagcg atgggtcggt gcaacatgcg ggatgatgtt
     6001 gcgccacacg taggaggagg tcgggttacc gtgcaggaac agcacagggg tgccatcgcg
     6061 cggaccaaca tcgacgtagt gcatgcgctc gcccaggact tccacataat gggggtcgaa
     6121 tggaaagcca gtaccgattt cggatccgag cgagaggcca acaaaggcca cgacggccag
     6181 taatatgcat aacttcatgg tggcggagcc tgcttttttg tacaaacttg tttcgaacga
     6241 aaggcccgga gatgaggaag aggagaacag cgcggcagac gtgcgctttt gaagcgtgca
     6301 gaatgccggg cctccggagg accttcgggc gcccgccccg cccctgagcc cgcccctgag
     6361 cccgcccccg gacccacccc ttcccagcct ctgagcccag aaagcgaagg agcaaagctg
     6421 ctattggccg ctgccccaaa ggcctacccg cttccattgc tcagcggtgc tgtccatctg
     6481 cacgagacta gtgagacgtg ctacttccat ttgtcacgtc ctgcacgacg cgagctgcgg
     6541 ggcggggggg aacttcctga ctaggggagg agtagaaggt ggcgcgaagg ggccaccaaa
     6601 gaacggagcc ggttggcgcc taccggtgga tgtggaatgt gtgcgaggcc agaggccact
     6661 tgtgtagcgc caagtgccca gcggggctgc taaagcgcat gctccagact gccttgggaa
     6721 aagcgcctcc cctacccata gggataacag ggtaatatag gcggccgcgt agatggcgcg
     6781 ccggtacctt aattaaccaa actggatctc tgctgtccct gtaataaacc cgaaaatttt
     6841 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
     6901 ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga
     6961 atactgccat ttgtctgcag aattggcgca cgcagtgccg atccgttcac taatcgaatg
     7021 gatctgtctc tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc
     7081 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     7141 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     7201 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     7261 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     7321 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     7381 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     7441 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     7501 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     7561 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     7621 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     7681 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     7741 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     7801 ttatataatt cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc
     7861 ggccggccgc ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct atcttgtcta
     7921 aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg
     7981 tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca
     8041 gtatttgtct acagccttct gatgtttcta acaggccagg attaactgcg aatcgttcta
     8101 gctccctgct tgcccatact atatgtttta atttatattt tttctttccc cctggcctta
     8161 accgaatttt ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc
     8221 catctctctc cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc
     8281 cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg
     8341 gtttcccttt cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga
     8401 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     8461 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     8521 gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa aacgcgctgc
     8581 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     8641 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     8701 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     8761 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     8821 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     8881 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     8941 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     9001 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     9061 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     9121 ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat
     9181 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     9241 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     9301 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     9361 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     9421 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     9481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     9541 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     9601 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     9661 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     9721 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     9781 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     9841 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     9901 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     9961 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
    10021 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
    10081 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
    10141 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
    10201 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
    10261 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
    10321 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
    10381 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
    10441 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
    10501 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
