pBS-EF1afull-H2B-mCherry-P2A-Fluc

LOCUS        pBS-EF1afull-H2B-mCherry-P2A-Fluc       11927 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral transfer genome encoding H2B-mCherry-P2A-Fluc driven by a
            full-length EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11927)
  AUTHORS   Strebinger D, Frangieh CJ, Friedrich MJ, Faure G, Macrae RK, Zhang F
  TITLE     Cell type-specific delivery by modular envelope design.
  JOURNAL   Nat Commun. 2023 Aug 23;14(1):5141. doi: 10.1038/s41467-023-40788-8.
  PUBMED    37612276
REFERENCE   2  (bases 1 to 11927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-08-23"; volume: "14(1):5141. doi"; pages: " 
            10.1038/s41467-023-40788-8"
COMMENT     Sequence Label: pBS-EF1afull-H2B-mCherry-P2A-Fluc
FEATURES             Location/Qualifiers
     source          1..11927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             119..299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(325..342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    331..555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      601..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(688..707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     898..919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1038..1058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1043..1372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1223..1358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1285..1304
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1420..1467
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1486..1860
                     /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    1990..2111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2027..2046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2081..2100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2160..2176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2160..2176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2173..2195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2184..2200
                     /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(2208..2214,2215..2232,2233..2238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2253..2274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2391..2408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2562..3150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2642..2661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3321..4112,4113..4181))
                     /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     3944..3963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4182..4286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4361..4380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4552..4931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4933..5131
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5086..5106
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5149..5329
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5376..5501
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5994..6227
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6412..6456
                     /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    6754..6871
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6921..7132
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          7133..8071
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8028..8048
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             8089..8466
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             8473..9180
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8618..8636)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8886..8905
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9196..9252
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             9256..10908
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    10933..11521
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10986..11006)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11404..11415)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11524..11540)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11525..11541)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
       61 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
      121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      301 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
      361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
      421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
      481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
      541 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
      601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      961 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     1321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     1381 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     1441 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     1501 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     1561 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     1621 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     1681 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     1741 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     1801 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     1861 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1921 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1981 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2101 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     2161 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2221 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2281 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2341 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3421 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4261 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     4321 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     4381 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     4441 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     4501 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4561 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4621 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4681 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4741 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4801 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4861 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4921 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4981 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5041 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5101 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
     5161 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5221 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5281 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     5341 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     5401 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     5461 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     5521 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     5581 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     5641 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     5701 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     5761 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     5821 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     5881 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     5941 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     6001 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     6061 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     6121 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     6181 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     6241 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     6301 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     6361 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     6421 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     6481 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     6541 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     6601 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     6661 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     6721 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     6781 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     6841 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     6901 agtttggtta attagctagc gggcagagcg cacatcgccc acagtccccg agaagttggg
     6961 gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     7021 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     7081 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     7141 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     7201 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     7261 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     7321 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     7381 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     7441 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     7501 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     7561 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     7621 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     7681 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     7741 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     7801 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     7861 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     7921 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     7981 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     8041 agtggttcaa agtttttttc ttccatttca gtctagagcg ctgccaccat gccagagcca
     8101 gcgaagtctg ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag
     8161 aaaggcggca agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag
     8221 gttctgaagc aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaat
     8281 tcgtttgtga acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac
     8341 aacaagcgct cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct
     8401 ggggagttgg ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc
     8461 gctaagccta ggatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     8521 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     8581 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     8641 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     8701 tacgtgaagc accccgccga cattcccgac tacttgaagc tgtccttccc cgagggcttc
     8761 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     8821 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     8881 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     8941 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     9001 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     9061 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     9121 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     9181 ggatccggca cgcgtgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
     9241 aatcctggac cgtccatgga agatgccaaa aacattaaga agggcccagc gccattctac
     9301 ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg
     9361 gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac ctacgccgag
     9421 tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct gaatacaaac
     9481 catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc
     9541 ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg cgagctgctg
     9601 aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag
     9661 atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat ggatagcaag
     9721 accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt gccacccggc
     9781 ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat cgccctgatc
     9841 atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca ccgcaccgct
     9901 tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat ccccgacacc
     9961 gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac gctgggctac
    10021 ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct attcttgcgc
    10081 agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt tagcttcttc
    10141 gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat cgccagcggc
    10201 ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca cctaccaggc
    10261 atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac ccccgaaggg
    10321 gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac
    10381 ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt ccgtggcccc
    10441 atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat cgacaaggac
    10501 ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt cttcatcgtg
    10561 gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc cgaactggag
    10621 agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct gcccgacgac
    10681 gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac catgaccgag
    10741 aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct gcgcggtggt
    10801 gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc ccgcaagatc
    10861 cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaaga attcgatatc
    10921 aagcttatcg gtaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
    10981 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
    11041 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
    11101 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
    11161 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
    11221 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
    11281 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
    11341 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
    11401 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
    11461 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
    11521 catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag tagcaataca
    11581 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
    11641 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
    11701 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
    11761 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca
    11821 ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt
    11881 gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgctt
//