LOCUS       sequence-332981-       10177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA and EGFP tagged RPL22, puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10177)
  AUTHORS   Gregory JA, Hoelzli E, Abdelaal R, Braine C, Cuevas M, Halpern M, 
            Barretto N, Schrode N, Akbalik G, Kang K, Cheng E, Bowles K, Lotz S,
            Goderie S, Karch CM, Temple S, Goate A, Brennand KJ, Phatnani H
  TITLE     Cell Type-Specific In Vitro Gene Expression Profiling of Stem 
            Cell-Derived Neural Models.
  JOURNAL   Cells. 2020 Jun 5;9(6). pii: cells9061406. doi: 
            10.3390/cells9061406.
  PUBMED    32516938
REFERENCE   2  (bases 1 to 10177)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cells."; 
            date: "2020-06-5"; volume: "9(6). pii: cells9061406. doi"; pages: " 
            10.3390/cells9061406"
COMMENT     Sequence Label: P-Ef1a-RPL22-3XHA-P2A-EGFP-T2A-Puro
FEATURES             Location/Qualifiers
     source          1..10177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        86..133
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             152..526
                     /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    656..777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(693..712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     747..766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(826..842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(826..842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(839..861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    850..866
                     /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(874..880,881..898,899..904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    919..940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1057..1074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1228..1816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1308..1327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1987..2778,2779..2847))
                     /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     2610..2629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2848..2952)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3027..3046)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3218..3597
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3599..3797
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3752..3772
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3815..3995
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4042..4167
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4660..4893
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5078..5122
                     /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    5420..5537
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5631..5842
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5861..5870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6266..6346
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             6362..6418
                     /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             6431..7141
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             6431..7141
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(6470..6491)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6731..6750)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7078..7099
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7151..7204
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7217..7810
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="EYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVT
                     ELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQ
                     QQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAP
                     RNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7707..7727
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7849..8437
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7902..7922)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8320..8331)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8440..8456)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8441..8457)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8962..9142
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9168..9185)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9174..9398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9444..9872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9531..9550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9741..9762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9881..9901)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(9886..10177,1..38)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(10066..10177,1..24)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10128..10147
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
       61 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
      121 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
      181 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
      241 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
      301 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
      361 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
      421 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
      481 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
      541 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
      601 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
      661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      781 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
      841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     2281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     2341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     3001 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3061 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     3121 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     3181 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
     3241 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     3301 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     3361 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     3421 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     3481 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     3541 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     3601 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     3661 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     3721 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     3781 gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg
     3841 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3901 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3961 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     4021 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     4081 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     4141 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     4201 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     4261 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     4321 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     4381 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     4441 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     4501 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     4561 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     4621 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     4681 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     4741 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     4801 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     4861 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     4921 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     4981 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     5041 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     5101 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     5161 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     5221 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     5281 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     5341 agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa
     5401 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     5461 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     5521 caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta
     5581 agctagctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     5641 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta
     5701 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     5761 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     5821 cgggtttgcc gccagaacac aggaccggtt aatctgcgct gccaccatgg ctcctgtgaa
     5881 aaagcttgtg gtgaaggggg gcaaaaaaaa gaagcaagtt ctgaagttca ctcttgattg
     5941 cacccaccct gtagaagatg gaatcatgga tgctgccaat tttgagcagt ttttgcaaga
     6001 aaggatcaaa gtgaacggaa aagctgggaa ccttggtgga ggggtggtga ccatcgaaag
     6061 gagcaagagc aagatcaccg tgacatccga ggtgcctttc tccaaaaggt atttgaaata
     6121 tctcaccaaa aaatatttga agaagaataa tctacgtgac tggttgcgcg tagttgctaa
     6181 cagcaaagag agttacgaat tacgttactt ccagattaac caggacgaag aagaggagga
     6241 agacgaggat ggaggtggat caggataccc atacgatgtt ccagattacg cttatccata
     6301 tgatgtccca gactatgctt acccttacga cgttcctgac tacgccgaat tcggatctgg
     6361 cgccacgaat ttctcgctac tcaagcaggc cggtgatgtc gaggaaaacc ctggtcctat
     6421 ggtgtctaga agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     6481 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     6541 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     6601 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     6661 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     6721 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     6781 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     6841 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     6901 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     6961 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     7021 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     7081 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     7141 gggttcaggg gagggaagag gttccctgct cacatgcgga gacgtagaag agaatcccgg
     7201 acccatgacc tctagagagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     7261 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     7321 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     7381 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     7441 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     7501 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     7561 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     7621 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     7681 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     7741 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     7801 cggtgcctga ttaattaagc tagctaattc gatatcaagc ttatcgataa tcaacctctg
     7861 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     7921 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     7981 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     8041 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     8101 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     8161 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     8221 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     8281 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     8341 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     8401 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgagac
     8461 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     8521 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     8581 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     8641 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     8701 acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca
     8761 ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc
     8821 aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     8881 gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca catggcccga
     8941 gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9001 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9061 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9121 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     9181 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     9241 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     9301 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     9361 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     9421 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9481 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     9541 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     9601 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     9661 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     9721 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     9781 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     9841 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     9901 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     9961 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10021 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    10081 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    10141 gccgaggccg cctctgcctc tgagctattc cagaagt
//
