LOCUS       pFSynW_syt1_143-        9979 bp DNA     circular SYN 25-AUG-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9979)
  AUTHORS   nmehta22
  TITLE     Direct Submission
  JOURNAL   Exported Sep 12, 2023 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     primer_bind     2582..2624
                     /label=NC32 For IF
                     /note="color: black; sequence: 
                     GATCCAGTTTGGTTAATTAAagtgcaagtggttttaggaccag"
     promoter        2602..3048
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     3033..3092
                     /label=142F3
                     /note="color: black; sequence: 
                     gtgcctgagagcgcagGTACGGCTAGCGCCACCATGggaaagctccaatattcactgga
                     c; added: 2023-06-30"
     primer_bind     3042..3090
                     /label=Fragment.FOR
                     /note="color: black; sequence: 
                     agcgcagGTACGGCTAGCGCCACCATGggaaagctccaatattcactgg; added: 
                     2023-06-21"
     CDS             3066..3068
                     /codon_start=1
                     /label=rSyt1_FL
                     /note="color: #993366"
                     /translation="M"
     CDS             3069..3905
                     /codon_start=1
                     /label=rSyt1_FL
                     /note="color: #993366"
                     /translation="GKLQYSLDYDFQNNQLLVGIIQAAELPALDMGGTSDPYVKVFLLP
                     DKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSKHDIIGEFKVP
                     MNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLRYVPTAGKLTVVILEAKNLKKMDVG
                     GLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYNESFSFEVPFEQIQKVQVVVTVLDY
                     DKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK"
     misc_feature    3069..3905
                     /label=Syt1 C2AB 80-421
                     /note="color: #a6acb3"
     primer_bind     3296..3319
                     /label=Primer 1
                     /note="color: black; sequence: attaggtggcaaaaccctggtgat; 
                     added: 2020-09-16"
     primer_bind     complement(3296..3319)
                     /label=Primer 2
                     /note="color: black; sequence: atcaccagggttttgccacctaat; 
                     added: 2020-09-16"
     primer_bind     3885..3970
                     /label=msGFP-FLAG 60 F (EC13459)
                     /note="color: black; sequence: 
                     GCCATGCTGGCTGTCAAGAAGGGATCCGGAAGTTCGGGGTCCAGCGGTTCTTCAGGCAG
                     TTCCATGGATAGCACTGAGAGCCTGTT; added: 2020-09-21"
     primer_bind     complement(3886..3905)
                     /label=Syt1 C2AB 80 R
                     /note="color: black; sequence: 
                     ATAAATACCTAAGCTTttacttcttgacagccagcatgg; added: 2020-09-04"
     CDS             3906..3947
                     /label=GS Linker
                     /note="color: #993366; direction: RIGHT"
     misc_feature    3906..3911
                     /label=linker for pre-prolactin
                     /note="found in original sequence"
                     /note="color: #a6acb3"
     primer_bind     3946..3983
                     /label=DSTES
                     /note="Use together with BL14 to amplify msGFP, msYFP, and 
                     msCFP with the improved N-terminal codons from A24. 
                     Subclone the PCR product back into the same vector using 
                     BamHI and NotI."
                     /note="color: black; sequence: 
                     CCGGGATCCACCGGTCGCCACCATGGATAGCACTGAGAGCCTGTTCACCGGGGTGGTG"
     CDS             3948..4643
                     /codon_start=1
                     /label=msGFP
                     /note="color: #05fd14"
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHG"
     CDS             3948..3965
                     /codon_start=1
                     /product="""An N-terminal MDSTES peptide in place of 
                     MVSKGEE """"In particular, the widely used red FP mCherry 
                     contains the N- and C-terminal peptides of EGFP (12), and 
                     we find that the N-terminal peptide of EGFP can promote 
                     cytotoxicity of mCherry during bacterial expression.""
                       A photostable monomeric superfolder GFPFernando Valbuena,
                     Ivy Fizgerald, Rita L. Strack, Neal Andruska, Luke Smith, 
                     Benjamin S. Glick 
                     "
                     /label=N-terminus mutation
                     /note="color: #993366"
                     /translation="MDSTES"
     primer_bind     3951..3974
                     /label=new N GFP lin F
                     /note="color: black; sequence: 
                     GCCATTTGCTAAACCTGTTAGAGCTGATAGCACTGAGAGCCTGTTCACC"
     CDS             4035..4037
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850."
                     /label=superfolder mutation (1/8) S30R
                     /note="color: #993366"
                     /translation="R"
     CDS             4062..4064
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850."
                     /label=superfolder mutation (2/8) Y39N
                     /note="color: #993366"
                     /translation="N"
     primer_bind     complement(4075..4097)
                     /label=msGFP N term R
                     /note="color: black; sequence: GGTGGTGCAGATGAACTTCAGGG; 
                     added: 2020-08-31"
     CDS             4242..4244
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850."
                     /label=superfolder mutation (3/8) F99T
                     /note="color: #993366"
                     /translation="T"
     CDS             4260..4262
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850."
                     /label=superfolder mutation (4/8) N105T
                     /note="color: #993366"
                     /translation="T"
     CDS             4380..4382
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850. 
                     Reversion of this mutation (F145Y) restores photostability 
                     of superfolding mutant with only a minor effect on folding 
                     Pedelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS.
                     Engineering and characterization of a superfolder green 
                     fluorescent protein. Nat Biotechnol 2006;24:79–88.
                     "
                     /label=superfolder mutation (5/8) Y145F
                     /note="color: #993366"
                     /translation="F"
     CDS             4404..4406
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850."
                     /label=superfolder mutation (6/8) M153T
                     /note="color: #993366"
                     /translation="T"
     primer_bind     complement(4427..4451)
                     /label=BQ13
                     /note="color: black; sequence: GCGGATCTTGAAGTTGGCCTTGATG"
     CDS             4434..4436
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850"
                     /label=superfolder mutation (7/8) V163A
                     /note="color: #993366"
                     /translation="A"
     CDS             4458..4460
                     /codon_start=1
                     /product="Shagin DA, Barsova EV, Yanushevich YG, Fradkov 
                     AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers 
                     A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like 
                     proteins as ubiquitous metazoan superfamily: evolution of 
                     functional features and structural complexity. Mol Biol 
                     Evol 2004;21(5):841–850."
                     /label=superfolder mutation (8/8) I171V
                     /note="color: #993366"
                     /translation="V"
     primer_bind     4518..4533
                     /label=msGFP C term F
                     /note="color: black; sequence: GGCCCCGTGCTGCTGC; added: 
                     2020-08-31"
     CDS             4563..4565
                     /codon_start=1
                     /product="Costantini LM, Fossati M, Francolini M, Snapp EL.
                     Assessing the tendency of fluorescent proteins to 
                     oligomerize under physiologic conditions. Traffic 
                     2012;13(5):643–649.
                     Zacharias DA, Violin JD, Newton AC, Tsien RY. Partitioning 
                     of lipid-modified monomeric GFPs into membrane microdomains
                     of live cells. Science 2002;296:913–916.
                     "
                     /label=monomerizing mutation A206K
                     /note="color: #993366"
                     /translation="K"
     primer_bind     complement(4616..4675)
                     /label=GFPR3
                     /note="color: black; sequence: 
                     TGATTATCGATAAGCTTGATATCGAATTCttaGCCGTGAGTGATCCCGGCGGCGGTCAC
                     G; added: 2023-06-30"
     primer_bind     complement(4626..4667)
                     /label=msGFP FLAG 60 R (EC13899)
                     /note="color: black; sequence: 
                     GATAAGCTTGATATCGAATTTTACTTGTCGTCATCGTCTTTGTAGTCTGAAGAACCGCC
                     GTGAGTGATCCCGGC; added: 2020-09-21"
     primer_bind     complement(4626..4643)
                     /label=BL14
                     /note="This oligo (used with BL13) will replace the linker 
                     MDELYK to GSSGSS in monomeric superfolder GFP, CFP and 
                     YFP."
                     /note="color: black; sequence: 
                     GATCTAGAGTCGCGGCCGCTTTAcccgctggagccggagctGCCGTGAGTGATCCCGGC
                     "
     primer_bind     complement(4628..4664)
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     AAGCTTGATATCGAATTCTTaGCCGTGAGTGATCCCG; added: 2023-06-21"
     primer_bind     complement(4633..4643)
                     /label=BL24
                     /note="This oligo is used to amplify msfFPs (Green, Cyan, 
                     Yellow) from the pMSGFP vector into the NT-HT vector for 
                     the Tub1 fussion.  The cut sites are BspEI for the 
                     N-terminal and SacI for the C-terminal. Use this oligo with
                     BL23."
                     /note="color: black; sequence: 
                     GGTTCCAGAGGGAGCACCGGAGCTCGTcccgctggagccggagctGCCGTGAGTGA"
     CDS             4638..4640
                     /codon_start=1
                     /product="Tsien RY. The green fluorescent protein. Annu Rev
                     Biochem 1998;67:509–544."
                     /label=monomeric mutation L231H
                     /note="color: #993366"
                     /translation="H"
     misc_feature    4671..5259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(5142..5153)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 5142"
                     /translation="IEGR"
     primer_bind     complement(5262..5278)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     LTR             5784..5964
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    5996..6220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      6266..6694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        6708..7037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      6888..7023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     promoter        7085..7132
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
                     /note="color: #ffffff; direction: RIGHT"
     CDS             7151..7525
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /note="color: #ccffcc"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7655..7776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(7825..7841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    7849..7865
                     /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)."
                     /note="color: #31849b"
     promoter        complement(7873..7903)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 7873 .. 7879 / #ffffff / -10
                        2: 7880 .. 7897 / #ffffff
                        3: 7898 .. 7903 / #ffffff / -35"
     protein_bind    7918..7939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(8227..8815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(8986..9846)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 8986 .. 9777 / #ccffcc
                        2: 9778 .. 9846 / #ccffcc / signal sequence
                     Cleavage site after base 9778"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9847..9951)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aagtgcaagt ggttttagga ccaggatgag gcggggtggg
     2641 ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa
     2701 attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca
     2761 ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc
     2821 cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgtaaac tccccttccc
     2881 ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc
     2941 gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct
     3001 cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcaggt acggctagcg
     3061 ccaccatggg aaagctccaa tattcactgg actatgactt ccagaataac cagctgttgg
     3121 tgggaatcat ccaggctgct gaactgcccg ccctggacat ggggggtaca tccgatccat
     3181 acgtcaaagt cttcctgctg cctgacaaaa agaagaaatt tgagactaaa gtccaccgga
     3241 aaaccctcaa tccagtcttc aatgaacaat ttactttcaa ggtaccctac tcggaattag
     3301 gtggcaaaac cctggtgatg gctgtgtatg actttgatcg cttctccaag cacgacatca
     3361 tcggagagtt caaagttcct atgaacaccg tggattttgg ccatgtgacc gaggagtggc
     3421 gcgatctcca gagcgctgag aaagaagagc aagagaaact gggtgacatc tgcttctccc
     3481 tccgctacgt ccctactgcc ggcaaactga ctgttgtcat tctggaagcc aagaacctga
     3541 agaagatgga tgtgggtggc ttatctgatc cctacgtgaa gattcacctg atgcagaacg
     3601 gtaagaggct gaagaagaaa aagacgacga ttaagaagaa cacactcaac ccctactaca
     3661 acgagtcctt cagctttgaa gttccgttcg agcaaatcca gaaagtgcaa gtggtggtaa
     3721 ctgttttgga ctatgacaag attggcaaga acgacgccat cggaaaagtc ttcgttggtt
     3781 acaacagcac tggggcggag ctgcgacact ggtcagacat gctggccaac ccccggcgac
     3841 ccatcgcaca gtggcacact ctgcaggtag aggaggaggt tgatgccatg ctggctgtca
     3901 agaagggatc cggaagttcg gggtccagcg gttcttcagg cagttccatg gatagcactg
     3961 agagcctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4021 acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     4081 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4141 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     4201 agtccgccat gcccgaaggc tacgtccagg agcgcaccat caccttcaag gacgacggca
     4261 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4321 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4381 tcaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     4441 tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4501 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4561 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4621 ccgccgccgg gatcactcac ggctaagaat tcgatatcaa gcttatcgat aatcaacctc
     4681 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4741 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4801 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4861 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4921 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4981 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5041 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     5101 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     5161 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     5221 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag
     5281 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     5341 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     5401 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5461 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     5521 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
     5581 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
     5641 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
     5701 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
     5761 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5821 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5881 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5941 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
     6001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6181 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     6241 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6481 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6601 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6661 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     6721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6961 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7021 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     7081 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     7141 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     7201 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     7261 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     7321 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     7381 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     7441 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     7501 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     7561 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     7621 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     7681 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7741 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     7801 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7861 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7921 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7981 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     8041 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     8101 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     8161 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     8221 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     8281 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     8341 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     8401 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     8461 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8521 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8581 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8641 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8701 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8761 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8821 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8881 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8941 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     9001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     9061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     9121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     9181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     9241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     9301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9601 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9841 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9901 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9961 ccgaaaagtg ccacctgac
//
