LOCUS       sequence-403585-       11505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2nd generation lentiviral transfer plasmid for engineering cells to 
            express human CD4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11505)
  AUTHORS   Hamilton JR, Chen E, Perez BS, Sandoval Espinoza CR, Kang MH, 
            Trinidad M, Ngo W, Doudna JA
  TITLE     In vivo human T cell engineering with enveloped delivery vehicles.
  JOURNAL   Nat Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z.
  PUBMED    38212493
REFERENCE   2  (bases 1 to 11505)
  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: "Nat 
            Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z."
COMMENT     Sequence Label: pJRH-1364 LV EF1a-CD4 IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..11505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(125..144)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        310..689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        691..889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     844..864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             907..1087
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1134..1259
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1752..1985
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2170..2214
                     /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    2512..2629
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2739..3916
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2970..3907
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3864..3884
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    5354..5927
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5377..5929
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5521..5538)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5748..5767
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5941..5943,5944..5946,5947..6660)
                     /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(5941..5943,5944..5946,5947..6657)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5986..6007)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6247..6266)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6594..6615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6675..7263
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6728..6748)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7146..7157)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7335..7568
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7594..7611)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7600..7824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7870..8298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7957..7976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8167..8188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8307..8327)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8312..8641
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8492..8627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8554..8573
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8689..8736
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8755..9129
                     /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    9259..9380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9296..9315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9350..9369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9429..9445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9429..9445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9442..9464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9453..9469
                     /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(9477..9483,9484..9501,9502..9507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9522..9543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9660..9677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9831..10419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9911..9930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10590..11381,11382..11450))
                     /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     11213..11232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(11451..11505,1..50))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
       61 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
      121 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
      181 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
      241 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      301 ccagatattg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      361 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      481 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      541 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      601 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      661 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      721 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      781 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      841 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
      901 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      961 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1021 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1081 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     1141 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1201 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1261 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1321 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1381 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1441 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1501 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1561 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     1621 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1681 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1741 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1801 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1861 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1921 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1981 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     2041 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     2101 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     2161 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     2221 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     2281 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     2341 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     2401 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     2461 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
     2521 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2581 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     2641 cagggacagc agagatccag tttggttaat taaggtacca tcgatcttaa gtcgcgactc
     2701 gagaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     2761 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2821 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2881 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2941 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     3001 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccactggc tgcagtacgt
     3061 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
     3121 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
     3181 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
     3241 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
     3301 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
     3361 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
     3421 ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc
     3481 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
     3541 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
     3601 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
     3661 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
     3721 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     3781 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     3841 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     3901 atttcaggtg tcgtgaaccg gttctagagc gctgccacca tgaacagggg agtgccattt
     3961 cgccatctgc tcctggtgtt gcagctcgcg ctccttccgg cggctaccca gggtaaaaag
     4021 gtagtcctcg gaaaaaaagg cgacacagtg gaacttacct gcaccgcgag ccaaaagaag
     4081 tctattcagt ttcattggaa gaactcaaat cagatcaaga tacttggcaa ccaaggatct
     4141 tttctgacta agggaccttc aaaacttaat gacagggcag attctaggag atctttgtgg
     4201 gaccaaggga attttcctct tattatcaaa aatctcaaga tagaggacag tgacacatat
     4261 atctgtgagg tggaagatca aaaggaggaa gtgcaactgc tcgtcttcgg gctgacagcc
     4321 aacagcgata cgcacttgct tcaagggcag tctcttaccc ttaccttgga gtctccaccc
     4381 gggtcatccc cttcagttca gtgccggagc ccgcgcggta agaacataca aggaggcaag
     4441 acgttgtctg tgtcccagct tgagctccaa gatagcggga cctggacgtg tacagtcctc
     4501 cagaatcaaa agaaggtaga gtttaaaatc gatatagtcg tcctcgcctt ccagaaggct
     4561 agttccatag tatataaaaa ggaaggcgag caagtcgagt ttagttttcc cctggcgttt
     4621 accgtcgaga agttgactgg tagcggtgag ctttggtggc aagctgagag ggcaagttct
     4681 tcaaaatcct ggatcacatt tgatctcaag aataaagagg ttagcgttaa acgggtgacg
     4741 caagacccga aactccaaat gggaaagaaa ttgcctctgc accttactct tccgcaagct
     4801 cttccccaat acgctggtag tggtaatttg accttggcac tggaagcaaa aacaggcaag
     4861 ctccaccagg aggtaaattt ggtagttatg cgcgcgactc aacttcaaaa gaacttgact
     4921 tgcgaggttt gggggcccac gagcccaaag cttatgcttt ctctgaagct tgagaataaa
     4981 gaggccaagg tcagtaagcg agaaaaggcg gtgtgggttt tgaaccccga agcagggatg
     5041 tggcaatgcc tcctcagcga ctccggtcaa gtactcttgg aatccaacat caaggttctc
     5101 cccacctgga gtacgccagt acagcctatg gcactcattg tactgggggg cgttgcgggt
     5161 ctcttgcttt ttatcggttt gggtatcttc ttttgtgtaa ggtgtcggca tagacggcga
     5221 caggcagaac gcatgtccca aataaaacgc cttctctctg aaaagaagac ctgccaatgc
     5281 ccgcatagat tccagaagac ttgctcccca atataaacgc gttaagtcga caaatgacaa
     5341 gtaagcggcc gcgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     5401 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     5461 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     5521 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     5581 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     5641 gcgacaggtg cctctgcggc ccaaagccac gtgtataaga tacacctgca aaggcggcac
     5701 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     5761 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     5821 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     5881 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacacat
     5941 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     6001 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     6061 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     6121 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6181 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     6241 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     6301 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     6361 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     6421 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     6481 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     6541 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     6601 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag
     6661 cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     6721 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     6781 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     6841 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     6901 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     6961 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     7021 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     7081 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     7141 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     7201 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     7261 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     7321 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     7381 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     7441 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7501 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7561 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     7621 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     7681 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     7741 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     7801 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     7861 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     7921 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     7981 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     8041 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     8101 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     8161 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     8221 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     8281 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     8341 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     8401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     8461 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     8521 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     8581 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     8641 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     8701 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     8761 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     8821 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     8881 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     8941 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     9001 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     9061 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     9121 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     9181 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     9241 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     9301 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     9361 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     9421 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     9481 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     9541 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     9601 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     9661 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     9721 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     9781 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     9841 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     9901 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     9961 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    10021 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    10081 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    10141 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    10201 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    10261 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    10321 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    10381 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    10441 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    10501 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    10561 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    10621 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    10681 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    10741 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    10801 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    10861 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    10921 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    10981 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    11041 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    11101 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    11161 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    11221 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    11281 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    11341 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    11401 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    11461 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagc
//
