LOCUS       sequence-335633-       12425 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For light regulated release of GluA1 from the endoplasmic reticulum.
            Driven by synapsin promoter. Thrombin site is included between GFP 
            and GluA1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12425)
  AUTHORS   Bourke AM, Schwartz SL, Bowen AB, Kleinjan MS, Winborn CS, Kareemo 
            DJ, Gutnick A, Schwarz TL, Kennedy MJ
  TITLE     zapERtrap: A light-regulated ER release system reveals unexpected 
            neuronal trafficking pathways.
  JOURNAL   J Cell Biol. 2021 Sep 6;220(9). pii: 212461. doi: 
            10.1083/jcb.202103186. Epub 2021 Jul 9.
  PUBMED    34241635
REFERENCE   2  (bases 1 to 12425)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2021-09-6"; volume: "220(9). pii: 212461. doi"; 
            pages: " 10.1083/jcb.202103186. Epub 2021 Jul 9"
COMMENT     Sequence Label: DHFR-GFP-th-GluA1
FEATURES             Location/Qualifiers
     source          1..12425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     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."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2632..3079
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             3195..3668
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             join(3690..3692,3693..4403)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3690..3692,3693..4403)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3732..3753)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3993..4012)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4340..4361
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4404..4421
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    7117..7705
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7170..7190)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7588..7599)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7708..7724)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7709..7725)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8230..8410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8436..8453)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8442..8666
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8712..9140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8799..8818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9009..9030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9149..9169)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9154..9483
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9334..9469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9396..9415
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9531..9578
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9597..9971
                     /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"
     polyA_signal    10101..10222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10138..10157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10192..10211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10271..10287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10271..10287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10284..10306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10295..10311
                     /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)."
     promoter        complement(join(10319..10325,10326..10343,10344..10349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10364..10385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10502..10519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10673..11261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10753..10772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11432..12223,12224..12292))
                     /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     12055..12074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12293..12397)
                     /gene="bla"
                     /label=AmpR promoter
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 agtgtctaga ctgcagaggg ccctgcgtat gagtgcaagt
     2641 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc
     2701 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag
     2761 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc
     2821 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc
     2881 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc
     2941 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct
     3001 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt
     3061 gtcgtgcctg agagcgcagt cgagaaacca ccatgccgta catctttgcc tttttctgca
     3121 ccggttttct aggtgcggtt gtgggtgcca atgcgcgagg tggaggcggt ggaagagggg
     3181 gtggaggaac gcgtatcagt ctgattgcgg cgttagcggt agatcgcgtt atcggcatgg
     3241 aaaacgccat gccgtggaac ctgcctgccg atctcgcctg gtttaaacgc aacaccttaa
     3301 ataaacccgt gattatgggc cgccatacct gggaatcaat cggtcgtccg ttgccaggac
     3361 gcaaaaatat tatcctcagc agtcaaccgg gtacggacga tcgcgtaacg tgggtgaagt
     3421 cggtggatga agccatcgcg gcgtgtggtg acgtaccaga aatcatggtg attggcggcg
     3481 gtcgcgttta tgaacagttc ttgccaaaag cgcaaaaact gtatctgacg catatcgacg
     3541 cagaagtgga aggcgacacc catttcccgg attacgagcc ggatgactgg gaatcggtat
     3601 tcagcgaatt ccacgatgct gatgcgcaga actctcacag ctattgcttt gagattctgg
     3661 agcggcggtc tgccggatcc gcagagcagg tgagcaaggg cgaggagctg ttcaccgggg
     3721 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3781 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3841 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3901 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3961 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     4021 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     4081 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4141 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     4201 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4261 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     4321 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4381 tcggcatgga cgagctgtat aagcttgttc ctcgaggaag cagtgctggt agtttcccca
     4441 acaatatcca gatagggggg ttatttccaa accaacaatc acaggaacat gcggctttta
     4501 ggtttgcttt gtcacaactc acggagcccc ccaagctgct tccccagatc gatattgtga
     4561 acatcagcga cagctttgag atgacttacc gtttctgttc ccagttctcc aaaggagtct
     4621 atgccatctt tggattttat gaacgaagga ctgtcaacat gctgacctcc ttctgtgggg
     4681 ccctccatgt gtgcttcatt actccaagtt ttcctgttga cacatccaat caatttgtcc
     4741 ttcagctacg cccggaacta caggaagctc tcattagcat tatcgaccat tacaagtggc
     4801 aaacctttgt ctacatttat gatgctgacc ggggcctgtc agtcctgcag agagtcttgg
     4861 atacagccgc agagaagaac tggcaggtaa cggctgtcaa cattctgaca accaccgagg
     4921 aaggataccg gatgctcttt caggacctgg agaagaaaaa ggagaggctg gtggtggttg
     4981 actgtgaatc agaacgcctc aacgccatcc tgggccagat cgtgaagcta gaaaagaatg
     5041 gcatcgggta ccactacatc ctcgccaatc tgggcttcat ggacattgac ttaaataagt
     5101 tcaaggagag cggagccaat gtgacaggtt tccagctggt gaactacaca gacacgatcc
     5161 cagccagaat catgcagcaa tggaggacaa gtgactcccg agaccatacc agggtggact
     5221 ggaagaggcc aaagtacact tctgctctca cctatgatgg tgtcaaggtg atggctgagg
     5281 ccttccaaag cctgcggagg cagaggattg acatatcccg ccgggggaat gctggggact
     5341 gtctggctaa cccagctgtg ccctggggtc aagggatcga catccagaga gccctgcagc
     5401 aggtgcgctt cgaaggtttg acaggaaatg tgcagttcaa cgagaaaggg cgccggacca
     5461 attacaccct ccacgtgatc gaaatgaaac atgatggaat ccgaaagatt ggttactgga
     5521 atgaagacga taaatttgtc cccgcagcca ccgacgctca ggctggaggg gacaactcaa
     5581 gcgtccagaa taggacctac atcgtcacta ctatcctcga agatccttac gtgatgctta
     5641 aaaagaatgc caaccagttt gagggcaatg accgctatga gggctactgt gtggagctgg
     5701 ctgcagagat cgccaagcac gtgggctact cctaccgact tgagattgtc agcgacggca
     5761 aatatggagc ccgggatccc gacacaaagg cttggaatgg catggtggga gaactggtct
     5821 atggaagagc agacgtggct gtggctccct tgaccataac cttggtccgg gaggaagtca
     5881 tcgacttctc caagccattc atgagtttgg gaatctccat tatgattaag aagccacaga
     5941 agtccaagcc aggtgtcttc tcctttcttg accctttggc ctatgagatc tggatgtgta
     6001 tagtgtttgc ctacattgga gtgagcgtcg tcctcttcct ggtcagccgt ttcagcccct
     6061 acgaatggca cagcgaagag tttgaagagg gacgagacca gacaaccagt gaccagtcaa
     6121 atgagtttgg catattcaac agcctgtggt tctccctggg ggccttcatg cagcaaggat
     6181 gtgacatttc ccccaggtcc ctgtccggac gcatcgtcgg cggcgtctgg tggttcttca
     6241 ctttgatcat catctcctcg tacacagcca acctggctgc cttcctgact gtggagagga
     6301 tggtgtctcc cattgagagt gcagaggacc tggcaaagca gacggaaatt gcttatggga
     6361 cattggaagc aggctccact aaggagttct tcaggagatc taaaatcgct gtgtttgaga
     6421 agatgtggac atacatgaag tctgcagaac catccgtgtt tgttcggacc acagaggaag
     6481 gcatgatcag agtgagaaaa tctaaaggca aatacgccta cctcctggag tccaccatga
     6541 atgagtatat tgagcaacga aagccctgtg acaccatgaa agtgggaggt aacttggatt
     6601 ccaaaggcta tggcattgcg acacccaagg ggtccgccct gagaaatcca gtaaacctgg
     6661 cagtgttaaa actgaacgag caggggcttt tggacaaatt gaaaaacaaa tggtggtacg
     6721 acaagggcga gtgcggcagc gggggaggtg actccaagga caagaccagc gctttgagcc
     6781 tcagcaatgt ggcaggcgtg ttctacatcc tgattggagg gctgggactg gccatgctgg
     6841 ttgccttaat cgagttctgc tacaaatccc gtagcgagtc gaagcggatg aagggtttct
     6901 gtttgatccc acagcaatcc atcaatgaag ccatacggac atcgaccctc ccccggaaca
     6961 gtggggcagg agccagcgga ggaggcggca gtggagagaa tggccgggtg gtcagccagg
     7021 acttccccaa gtccatgcaa tccattccct gcatgagtca cagttcaggg atgcccttgg
     7081 gagccacagg attgtaacga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     7141 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     7201 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     7261 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     7321 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     7381 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     7441 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     7501 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     7561 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     7621 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     7681 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat
     7741 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     7801 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     7861 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     7921 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     7981 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     8041 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     8101 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     8161 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     8221 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     8281 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     8341 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     8401 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     8461 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     8521 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     8581 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     8641 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     8701 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     8761 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     8821 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     8881 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     8941 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9001 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9061 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     9121 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     9181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     9241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     9301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     9361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     9421 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     9481 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     9541 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     9601 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     9661 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     9721 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     9781 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     9841 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     9901 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     9961 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
    10021 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
    10081 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
    10141 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
    10201 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
    10261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    10321 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    10381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    10441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    10501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    10561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    10621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    10681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    10741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    10801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    10861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    10921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    10981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    11041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    11101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    11161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    11221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    11281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    11341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    11401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    11461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    11521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    11581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    11641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    11701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    11761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    11821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    11881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    11941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    12001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    12061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    12121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    12181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    12241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    12301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    12361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    12421 ctgac
//
