LOCUS       sequence-425601-       13618 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector for the expression of MCAM-GFP fusion protein in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13618)
  AUTHORS   Miller SG, Hoh M, Ebmeier CC, Tay JW, Ahn NG
  TITLE     Cooperative polarization of MCAM/CD146 and ERM family proteins in 
            melanoma.
  JOURNAL   Mol Biol Cell. 2023 Dec 20:mbcE23060255. doi: 
            10.1091/mbc.E23-06-0255.
  PUBMED    38117590
REFERENCE   2  (bases 1 to 13618)
  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: "Mol Biol 
            Cell. 2023 Dec 20:mbcE23060255. doi: 10.1091/mbc.E23-06-0255."
COMMENT     Sequence Label: MCAM-GFP
FEATURES             Location/Qualifiers
     source          1..13618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             831..1011
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1058..1183
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1675..1908
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2093..2137
                     /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    2411..2528
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2584..3746
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2815..3737
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3694..3714
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3771..3795
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(5788..5790,5791..5793,5794..6507)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5788..5790,5791..5793,5794..6504)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5833..5854)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6094..6113)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6441..6462
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6568..6592)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(6648..7222,7223..7225,7226..7234)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6815..6832)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7042..7061
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7260..8276
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             7260..8276
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    8312..8900
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8365..8385)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8783..8794)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9419..9599
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9625..9642)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9631..9855
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9901..10329
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9988..10007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10198..10219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10338..10358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10343..10672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10523..10658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10585..10604
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10720..10767
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10786..11160
                     /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    11290..11411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11327..11346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11381..11400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11460..11476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11460..11476)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11473..11495)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11484..11500
                     /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(11508..11514,11515..11532,11533..11538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11553..11574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11691..11708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11862..12450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11942..11961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12621..13412,13413..13481))
                     /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     13244..13263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13482..13586)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
      841 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      901 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      961 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     1021 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     1081 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     1141 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     1201 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     1261 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     1321 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     1381 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     1441 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     1501 taagaccacc gcacagcaag ggccgctgat cttcagacct ggaggaggag atatgaggga
     1561 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1621 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1681 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1741 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1801 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1861 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1921 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1981 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     2041 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     2101 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     2161 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     2221 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     2281 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     2341 tggagagaga gacagagaca gatccattcg attagtgaac ggatctacaa atggcagtat
     2401 tcatccacaa ttttaaaaga aaagggggga ttgggaggta cagtgcaggg gaaagaatag
     2461 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     2521 aaaattttcg ggtttattac agggacagca gaaattcact ttgaattaat tcaagcttcg
     2581 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     2641 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     2701 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     2761 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     2821 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     2881 attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc
     2941 cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3001 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3061 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3121 taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3181 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     3241 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     3301 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     3361 aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg
     3421 gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg
     3481 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct
     3541 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     3601 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     3661 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     3721 tttttcttcc atttcaggtg tcgtgaacac gctaccggtc tcgagaattg acaagtttgt
     3781 acaaaaaagc aggctccgct agcgctaccg gactcagatc tcgagctcaa gcttatgggg
     3841 cttcccaggc tggtctgcgc cttcttgctc gccgcctgct gctgctgtcc tcgcgtcgcg
     3901 ggtgtgcccg gagaggctga gcagcctgcg cctgagctgg tggaggtgga agtgggcagc
     3961 acagcccttc tgaagtgcgg cctctcccag tcccaaggca acctcagcca tgtcgactgg
     4021 ttttctgtcc acaaggagaa gcggacgctc atcttccgtg tgcgccaggg ccagggccag
     4081 agcgaacctg gggagtacga gcagcggctc agcctccagg acagaggggc tactctggcc
     4141 ctgactcaag tcacccccca agacgagcgc atcttcttgt gccagggcaa gcgccctcgg
     4201 tcccaggagt accgcatcca gctccgcgtc tacaaagctc cggaggagcc aaacatccag
     4261 gtcaaccccc tgggcatccc tgtgaacagt aaggagcctg aggaggtcgc tacctgtgta
     4321 gggaggaacg ggtaccccat tcctcaagtc atctggtaca agaatggccg gcctctgaag
     4381 gaggagaaga accgggtcca cattcagtcg tcccagactg tggagtcgag tggtttgtac
     4441 accttgcaga gtattctgaa ggcacagctg gttaaagaag acaaagatgc ccagttttac
     4501 tgtgagctca actaccggct gcccagtggg aaccacatga aggagtccag ggaagtcacc
     4561 gtccctgttt tctacccgac agaaaaagtg tggctggaag tggagcccgt gggaatgctg
     4621 aaggaagggg accgcgtgga aatcaggtgt ttggctgatg gcaaccctcc accacacttc
     4681 agcatcagca agcagaaccc cagcaccagg gaggcagagg aagagacaac caacgacaac
     4741 ggggtcctgg tgctggagcc tgcccggaag gaacacagtg ggcgctatga atgtcagggc
     4801 ctggacttgg acaccatgat atcgctgctg agtgaaccac aggaactact ggtgaactat
     4861 gtgtctgacg tccgagtgag tcccgcagcc cctgagagac aggaaggcag cagcctcacc
     4921 ctgacctgtg aggcagagag tagccaggac ctcgagttcc agtggctgag agaagagaca
     4981 ggccaggtgc tggaaagggg gcctgtgctt cagttgcatg acctgaaacg ggaggcagga
     5041 ggcggctatc gctgcgtggc gtctgtgccc agcatacccg gcctgaaccg cacacagctg
     5101 gtcaacgtgg ccatttttgg ccccccttgg atggcattca aggagaggaa ggtgtgggtg
     5161 aaagagaata tggtgttgaa tctgtcttgt gaagcgtcag ggcacccccg gcccaccatc
     5221 tcctggaacg tcaacggcac ggcaagtgaa caagaccaag atccacagcg agtcctgagc
     5281 accctgaatg tcctcgtgac cccggagctg ttggagacag gtgttgaatg cacggcctcc
     5341 aacgacctgg gcaaaaacac cagcatcctc ttcctggagc tggtcaattt aaccaccctc
     5401 acaccagact ccaacacaac cactggcctc agcacttcca ctgccagtcc tcataccaga
     5461 gccaacagca cctccacaga gagaaagctg ccggagccgg agagccgggg cgtggtcatc
     5521 gtggctgtga ttgtgtgcat cctggtcctg gcggtgctgg gcgctgtcct ctatttcctc
     5581 tataagaagg gcaagctgcc gtgcaggcgc tcagggaagc aggagatcac gctgcccccg
     5641 tctcgtaaga gcgaacttgt agttgaagtt aagtcagata agctcccaga agagatgggc
     5701 ctcctgcagg gcagcagcgg tgacaagagg gctccgggag accagggaga gaaatacatc
     5761 gatctgaggc atggatccgg tggaggtatg gtgagcaagg gcgaggagct gttcaccggg
     5821 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     5881 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     5941 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     6001 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     6061 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     6121 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     6181 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     6241 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     6301 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     6361 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     6421 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     6481 ctcggcatgg acgagctgta caagtaagcg gccgctcgac ggtaccgcgg gcccgggatc
     6541 ggccgcgact ctagatcata atcagccacc cagctttctt gtacaaagtg gtttgattcg
     6601 acccgggatc gggccgcact agaggaattc cgcccctctc cccccccccc ctctccctcc
     6661 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     6721 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     6781 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     6841 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     6901 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     6961 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     7021 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     7081 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     7141 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     7201 ctttgaaaaa cacgatgata agcttgccac aacccgtacc aaagatggat agatccggaa
     7261 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     7321 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     7381 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     7441 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     7501 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     7561 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     7621 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     7681 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     7741 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     7801 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     7861 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     7921 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     7981 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     8041 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     8101 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     8161 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     8221 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaagcta
     8281 gagttaacat cgagggatca agcttatcga taatcaacct ctggattaca aaatttgtga
     8341 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     8401 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     8461 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     8521 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     8581 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     8641 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     8701 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     8761 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     8821 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     8881 cctttgggcc gcctccccgc atcgataccg tcgagaccta gaaaaacatg gagcaatcac
     8941 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     9001 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     9061 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     9121 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     9181 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa
     9241 gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt
     9301 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
     9361 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
     9421 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     9481 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     9541 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     9601 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     9661 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     9721 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     9781 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     9841 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     9901 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     9961 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
    10021 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
    10081 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
    10141 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
    10201 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
    10261 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
    10321 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
    10381 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
    10441 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
    10501 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
    10561 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
    10621 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
    10681 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
    10741 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
    10801 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
    10861 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
    10921 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
    10981 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
    11041 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
    11101 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
    11161 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
    11221 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
    11281 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
    11341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    11401 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
    11461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    11521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    11581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    11641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    11701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    11761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    11821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    11881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    11941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    12001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    12061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    12121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    12181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    12241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    12301 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    12361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    12421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    12481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    12541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    12601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    12661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    12721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    12781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    12841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    12901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    12961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    13021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    13081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    13141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    13201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    13261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    13321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    13381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    13441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    13501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    13561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcg
//
