LOCUS       Exported                8936 bp ds-DNA     circular SYN 24-NOV-2025
DEFINITION  synthetic DNA.
ACCESSION   .
VERSION     .
KEYWORDS    NEBuilder.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8936)
  TITLE     NEBuilder-generated Construct
  JOURNAL   Exported 3-OCT-2022 from NEBuilder https://nebuilder.neb.com
REFERENCE   2  (bases 1 to 8936)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 25, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     NEBuilder-generated oligos (UPPERCASE = gene-specific, lowercase = 
            overlap) pCDNA4TO-3xFLAG-Halo_fwd: GTCTAGAGGGCCCGTTTAAAC
                     pCDNA4TO-3xFLAG-Halo_fwd 3'Tm: 65.1  3'Ta: 66.1
                     pCDNA4TO-3xFLAG-Halo_rev: GGCCGCGCTCTGAAAGTAC
                     pCDNA4TO-3xFLAG-Halo_rev 3'Tm: 69.2  3'Ta: 66.1
                     AARS1_fwd: tgtactttcagagcgcggccggaagcgctATGGACTCTACTCTAACAG
                    AARS1_fwd 3'Tm: 56.6  3'Ta: 57.6
            AARS1_rev: tttaaacgggccctctagacTCAGTTCTTTACATCCCC
            AARS1_rev 3'Tm: 57.8  3'Ta: 57.6.
FEATURES             Location/Qualifiers
     source          1..8936
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..21
                     /label=pCDNA4TO-3xFLAG-Halo_fwd
                     /note="pCDNA4TO-3xFLAG-Halo_fwd"
                     /note="gene-specific Tm: 65.1  Ta: 66.1"
                     /note="gene-specific primer: GTCTAGAGGGCCCGTTTAAAC"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(33..50)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer.
                     Also called BGH reverse"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     BGH-rev (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     BGH-rev
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     polyA_signal    39..263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH
                     poly(A) signal (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; bGH
                     poly(A) signal (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a6acb3"
     rep_origin      309..737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     f1
                     ori (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; f1 
                     ori
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffff00"
     primer_bind     complement(396..415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     F1ori-R (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     F1ori-R
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     606..627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     F1ori-F (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     F1ori-F
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(746..766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     pBABE 3' (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     pBABE 3'
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     promoter        751..1080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     SV40
                     promoter (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     SV40
                     promoter (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffffff"
     rep_origin      931..1066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     SV40
                     ori (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     SV40 ori
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffff00"
     primer_bind     993..1012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     SV40pro-F (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0;
                     SV40pro-F (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        1128..1175
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     EM7
                     promoter (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; EM7
                     promoter (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffffff"
     CDS             1194..1568
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     BleoR (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     BleoR
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="/Transferred_Translation=MAKLTSAVPVLTARDVAGAVEFWTDRL
                     GFSRD 
                     FVEDDFAGVVRDDVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDAS 
                     GPAMTEIGEQPWGREFALRDPAGNCVHFVAEEQD"
                     /note="color: #993366"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1698..1819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     SV40
                     poly(A) signal (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     SV40
                     poly(A) signal (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a6acb3"
     primer_bind     complement(1735..1754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     SV40pA-R (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     SV40pA-R
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     1789..1808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     EBV-rev (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     EBV-rev
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(1868..1884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     M13
                     Reverse (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; M13
                     Reverse (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(1868..1884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     M13
                     rev (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; M13
                     rev
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(1881..1903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     M13/pUC Reverse (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     M13/pUC
                     Reverse (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    1892..1908
                     /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)."
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     lac
                     operator (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; lac
                     operator (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #31849b"
     promoter        complement(1916..1946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     lac
                     promoter (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; lac
                     promoter (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    1961..1982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     CAP
                     binding site (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; CAP
                     binding site (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #31849b"
     primer_bind     complement(2099..2116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     L4440 (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     L4440
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      complement(2270..2858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     ori
                     (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; ori
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffff00"
     primer_bind     complement(2350..2369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     pBR322ori-F (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0;
                     pBR322ori-F (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(3029..3889)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR
                     (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     AmpR (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="/Transferred_Translation=MSIQHFRVALIPFFAAFCLPVFAHPET
                     LVKVK 
                     DAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLG 
                     RRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTA 
                     FLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLI 
                     DWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGS 
                     QATMDERNRQIAEIGASLIKHW"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3652..3671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     Amp-R (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     Amp-R
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(3890..3994)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR
                     promoter (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     AmpR
                     promoter (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(4069..4088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     pRS-marker (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0;
                     pRS-marker (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: LEFT"
     enhancer        4260..4639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     CMV
                     enhancer (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; CMV
                     enhancer (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffffff"
     promoter        4640..4843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     CMV
                     promoter (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; CMV
                     promoter (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     4794..4814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     CMV-F (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     CMV-F
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: RIGHT"
     protein_bind    4845..4863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     tet
                     operator (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     tet
                     operator (Temp-Halo-DCTPP1)"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     tet
                     operator (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; tet
                     operator (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #31849b"
     protein_bind    4866..4884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     tet
                     operator (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     tet
                     operator (Temp-Halo-DCTPP1)"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     tet
                     operator (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; tet
                     operator (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #31849b"
     primer_bind     4894..4918
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     LNCX
                     (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     LNCX (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             5019..5084
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by
                     an enterokinase cleavage site"
                     /label=3xFLAG
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     3xFLAG (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     3xFLAG
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="/Transferred_Translation=DYKDHDGDYKDHDIDYKDDDDK"
                     /note="color: #cc99b2"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5094..5981
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     HaloTag(R) (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0;
                     HaloTag(R) (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="/Transferred_Translation=AEIGTGFPFDPHYVEVLGERMHYVDVG
                     PRDGT 
                     PVLFLHGNPTSSYVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAF 
                     IEALGLEEVVLVIHDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETF 
                     QAFRTTDVGRKLIIDQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPN 
                     ELPIAGEPANIVALVEEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAV 
                     DIGPGLNLLQEDNPDLIGSEIARWLSTLEISG"
                     /note="color: #cc99b2"
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     5888..5907
                     /label=5,888 F
                     /note="color: black; sequence: CATCGGCCCGGGTCTGAATC"
     CDS             5994..6014
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition
                     and cleavage site"
                     /label=TEV site
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Alternative_Match=100.00%; Adjusted-Interval-0:0/0;
                     TEV
                     site (Temp-Halo-DCTPP1)"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; TEV
                     site
                     (pCDNA4TO-3XFLAG Halo WT nsp1 )"
                     /note="/Transferred_From=pCDNA4TO-3XFLAG Halo WT nsp1 "
                     /note="/Transferred_Similarity=100.00%"
                     /note="/Transferred_Translation=EDLYFQS"
                     /note="color: #cc99b2"
                     /translation="EDLYFQS"
     primer_bind     6001..6048
                     /label=AARS1_fwd
                     /note="AARS1_fwd"
                     /note="gene-specific Tm: 56.6  Ta: 57.6"
                     /note="overlap: tgtactttcagagcgcggcc"
                     /note="spacer: ggaagcgct"
                     /note="gene-specific primer: ATGGACTCTACTCTAACAG"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(6002..6020)
                     /label=pCDNA4TO-3xFLAG-Halo_rev
                     /note="pCDNA4TO-3xFLAG-Halo_rev"
                     /note="gene-specific Tm: 69.2  Ta: 66.1"
                     /note="gene-specific primer: GGCCGCGCTCTGAAAGTAC"
                     /note="color: #a020f0; direction: LEFT"
     CDS             6030..8936
                     /codon_start=1
                     /gene="AARS1"
                     /gene_synonym="AARS; CMT2N; DEE29; EIEE29; HDLS2; TTD8"
                     /product="alanine--tRNA ligase, cytoplasmic"
                     /EC_number="6.1.1.7"
                     /label=AARS1
                     /note="alanine tRNA ligase 1, cytoplasmic; alanyl-tRNA 
                     synthetase, cytoplasmic; alaRS; renal carcinoma antigen 
                     NY-REN-42"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="/Primary_Match=100.00%; Adjusted-Interval-0:0/0; 
                     AARS1
                     CDS (AARS1 NM_001605 - AARS1 CDS)"
                     /note="/Transferred_From=AARS1 NM_001605 - AARS1 CDS"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/Transferred_Translation=MDSTLTASEIRQRFIDFFKRNEHTYVH
                     SSATI 
                     PLDDPTLLFANAGMNQFKPIFLNTIDPSHPMAKLSRAANTQKCIRAGGKHNDLDDVGK 
                     DVYHHTFFEMLGSWSFGDYFKELACKMALELLTQEFGIPIERLYVTYFGGDEAAGLEA 
                     DLECKQIWQNLGLDDTKILPGNMKDNFWEMGDTGPCGPCSEIHYDRIGGRDAAHLVNQ 
                     DDPNVLEIWNLVFIQYNREADGILKPLPKKSIDTGMGLERLVSVLQNKMSNYDTDLFV 
                     PYFEAIQKGTGARPYTGKVGAEDADGIDMAYRVLADHARTITVALADGGRPDNTGRGY 
                     VLRRILRRAVRYAHEKLNASRGFFATLVDVVVQSLGDAFPELKKDPDMVKDIINEEEV 
                     QFLKTLSRGRRILDRKIQSLGDSKTIPGDTAWLLYDTYGFPVDLTGLIAEEKGLVVDM 
                     DGFEEERKLAQLKSQGKGAGGEDLIMLDIYAIEELRARGLEVTDDSPKYNYHLDSSGS 
                     YVFENTVATVMALRREKMFVEEVSTGQECGVVLDKTCFYAEQGGQIYDEGYLVKVDDS 
                     SEDKTEFTVKNAQVRGGYVLHIGTIYGDLKVGDQVWLFIDEPRRRPIMSNHTATHILN 
                     FALRSVLGEADQKGSLVAPDRLRFDFTAKGAMSTQQIKKAEEIANEMIEAAKAVYTQD 
                     CPLAAAKAIQGLRAVFDETYPDPVRVVSIGVPVSELLDDPSGPAGSLTSVEFCGGTHL 
                     RNSSHAGAFVIVTEEAIAKGIRRIVAVTGAEAQKALRKAESLKKCLSVMEAKVKAQTA 
                     PNKDVQREIADLGEALATAVIPQWQKDELRETLKSLKKVMDDLDRASKADVQKRVLEK 
                     TKQFIDSNPNQPLVILEMESGASAKALNEALKLFKMHSPQTSAMLFTVDNEAGKITCL 
                     CQVPQNAANRGLKASEWVQQVSGLMDGKGGGKDVSAQATGKNVGCLQEALQLATSFAQ 
                     LRLGDVKN"
                     /note="color: #993366"
                     /db_xref="CCDS:CCDS32474.1"
                     /db_xref="GeneID:16"
                     /db_xref="HGNC:HGNC:20"
                     /db_xref="MIM:601065"
                     /protein_id="NP_001596.2"
                     /translation="MDSTLTASEIRQRFIDFFKRNEHTYVHSSATIPLDDPTLLFANAG
                     MNQFKPIFLNTIDPSHPMAKLSRAANTQKCIRAGGKHNDLDDVGKDVYHHTFFEMLGSW
                     SFGDYFKELACKMALELLTQEFGIPIERLYVTYFGGDEAAGLEADLECKQIWQNLGLDD
                     TKILPGNMKDNFWEMGDTGPCGPCSEIHYDRIGGRDAAHLVNQDDPNVLEIWNLVFIQY
                     NREADGILKPLPKKSIDTGMGLERLVSVLQNKMSNYDTDLFVPYFEAIQKGTGARPYTG
                     KVGAEDADGIDMAYRVLADHARTITVALADGGRPDNTGRGYVLRRILRRAVRYAHEKLN
                     ASRGFFATLVDVVVQSLGDAFPELKKDPDMVKDIINEEEVQFLKTLSRGRRILDRKIQS
                     LGDSKTIPGDTAWLLYDTYGFPVDLTGLIAEEKGLVVDMDGFEEERKLAQLKSQGKGAG
                     GEDLIMLDIYAIEELRARGLEVTDDSPKYNYHLDSSGSYVFENTVATVMALRREKMFVE
                     EVSTGQECGVVLDKTCFYAEQGGQIYDEGYLVKVDDSSEDKTEFTVKNAQVRGGYVLHI
                     GTIYGDLKVGDQVWLFIDEPRRRPIMSNHTATHILNFALRSVLGEADQKGSLVAPDRLR
                     FDFTAKGAMSTQQIKKAEEIANEMIEAAKAVYTQDCPLAAAKAIQGLRAVFDETYPDPV
                     RVVSIGVPVSELLDDPSGPAGSLTSVEFCGGTHLRNSSHAGAFVIVTEEAIAKGIRRIV
                     AVTGAEAQKALRKAESLKKCLSVMEAKVKAQTAPNKDVQREIADLGEALATAVIPQWQK
                     DELRETLKSLKKVMDDLDRASKADVQKRVLEKTKQFIDSNPNQPLVILEMESGASAKAL
                     NEALKLFKMHSPQTSAMLFTVDNEAGKITCLCQVPQNAANRGLKASEWVQQVSGLMDGK
                     GGGKDVSAQATGKNVGCLQEALQLATSFAQLRLGDVKN"
     primer_bind     6398..6417
                     /label=6,398 F
                     /note="color: black; sequence: CCAAGAGTTTGGCATTCCCA"
     primer_bind     6803..6824
                     /label=6,803 F
                     /note="color: black; sequence: TGACACTGACCTTTTTGTCCCT"
     primer_bind     7396..7415
                     /label=7,396 F
                     /note="color: black; sequence: GAGCTGGTGGGGAAGACCTC"
     misc_feature    8744
                     /label=T
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=T"
                     /note="color: #a6acb3"
     primer_bind     complement(join(8919..8936,1..20))
                     /label=AARS1_rev
                     /note="AARS1_rev"
                     /note="gene-specific Tm: 57.8  Ta: 57.6"
                     /note="overlap: tttaaacgggccctctagac"
                     /note="gene-specific primer: TCAGTTCTTTACATCCCC"
                     /note="color: #a020f0; direction: LEFT"
ORIGIN
        1 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
       61 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
      121 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
      181 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
      241 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
      301 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      361 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      421 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      481 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      541 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      601 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      661 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      721 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
      781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
      841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
      901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
      961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     1021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     1081 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     1141 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     1201 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     1261 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     1321 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     1381 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     1441 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     1501 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     1561 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     1621 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     1681 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1741 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1801 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     1861 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1921 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1981 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2041 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2101 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2161 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2221 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2281 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2341 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2401 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2461 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2521 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2581 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2641 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2701 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2761 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2821 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2881 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2941 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3001 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3061 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3121 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3181 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3241 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3301 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3361 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3421 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3481 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3541 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3601 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3661 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3721 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3781 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3841 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3901 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3961 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4021 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     4081 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     4141 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     4201 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     4261 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     4321 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4381 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4441 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4501 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     4561 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     4621 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     4681 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     4741 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     4801 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctcccta tcagtgatag
     4861 agatctccct atcagtgata gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc
     4921 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     4981 ccggactcta gcgtttaaac ttaagcttac caatgatgga ctacaaggac cacgacggcg
     5041 actacaagga ccacgacatc gactacaagg acgacgacga caagggggcg gccgcagaaa
     5101 tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag cgcatgcact
     5161 acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt aacccgacct
     5221 cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc tgcattgctc
     5281 cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc ttcgacgacc
     5341 acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca
     5401 ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca
     5461 aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa tggccagaat
     5521 ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag ctgatcatcg
     5581 atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag
     5641 tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc
     5701 gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag
     5761 aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg ggcaccccag
     5821 gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct aactgcaagg
     5881 ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac ctgatcggca
     5941 gcgagatcgc gcgctggctg tcgacgctcg agatttccgg cgagccaacc actgaggatc
     6001 tgtactttca gagcgcggcc ggaagcgcta tggactctac tctaacagca agtgaaatcc
     6061 ggcagcgatt tatagatttc ttcaagagga acgagcatac gtatgttcac tcgtctgcca
     6121 ccatcccatt ggatgacccc actttgctct ttgccaatgc aggcatgaac cagtttaaac
     6181 ccattttcct gaacacaatt gacccatctc accccatggc aaagctgagc agagctgcca
     6241 atacccagaa gtgcatccgg gctgggggca aacataatga cctggacgat gtgggcaagg
     6301 atgtctatca tcacaccttc ttcgagatgc tgggctcttg gtcttttgga gattacttta
     6361 aggaattggc atgtaagatg gctctggaac tcctcaccca agagtttggc attcccattg
     6421 aaagacttta tgttacttac tttggcgggg atgaagcagc tggcttagaa gcagatctgg
     6481 aatgcaaaca gatctggcaa aatttggggc tggatgacac caaaatcctc ccaggcaaca
     6541 tgaaggataa cttctgggag atgggtgaca cgggcccctg tggtccttgc agtgagatcc
     6601 actacgaccg gattggtggt cgggacgccg cacatcttgt caaccaggac gaccctaatg
     6661 tgctggagat ctggaacctt gtgttcatcc agtataacag ggaagctgat ggcattctga
     6721 aacctcttcc caagaaaagc attgacacag ggatgggcct ggaacgactg gtatctgtgc
     6781 tgcagaataa gatgtccaac tatgacactg acctttttgt cccttacttt gaagccattc
     6841 agaagggcac aggtgcccga ccatacactg ggaaagttgg tgctgaggat gccgatggga
     6901 ttgacatggc ctaccgggtg ctggctgacc acgctcggac catcactgtg gcactggctg
     6961 atggtggccg gcctgacaac acagggcgtg gatatgtgtt gagacggatt ctccgccgag
     7021 ctgtccgata cgcccatgaa aagctcaatg ccagcagggg cttctttgct acgttagtgg
     7081 atgttgtcgt ccagtccctg ggagatgcat ttcctgagct gaagaaggac ccagacatgg
     7141 tgaaggacat cattaatgaa gaagaggtgc agtttctcaa gactctcagc agagggcgtc
     7201 gcatcctgga caggaaaatt cagagcctgg gagacagcaa gaccattccc ggagacactg
     7261 cttggctcct ctatgacacc tatgggtttc cagtggatct gactggactg attgctgaag
     7321 agaagggcct ggtggtagac atggatggct ttgaagagga gaggaaactg gcccagctga
     7381 aatcacaggg caagggagct ggtggggaag acctcattat gctggacatt tacgctatcg
     7441 aagagctccg ggcacggggt ctggaggtca cagatgattc cccaaagtac aattaccatt
     7501 tggactccag tggtagctat gtatttgaga acacagtggc tacggtgatg gctctgcgca
     7561 gggagaagat gttcgtggaa gaggtgtcca caggccagga gtgtggagtg gtgctggaca
     7621 agacctgttt ctatgctgag caaggaggcc agatctatga cgaaggctac ctggtgaagg
     7681 tggatgacag cagtgaagat aaaacagagt ttacagtgaa gaatgctcag gtccgaggag
     7741 ggtatgtgct acacattgga accatctacg gtgacctgaa agtgggggat caggtctggc
     7801 tgtttattga tgagccccga cgaagaccca tcatgagcaa ccacacagct acgcacattc
     7861 tgaacttcgc cctgcgctca gtgcttgggg aagctgacca gaaaggctca ttggttgctc
     7921 ctgaccgcct cagatttgac tttactgcca agggagccat gtccacccaa cagatcaaga
     7981 aggctgaaga gattgctaat gagatgattg aggcagccaa ggccgtctat acccaggatt
     8041 gccccctggc agcagcgaaa gccatccagg gcctacgggc tgtgtttgat gagacctatc
     8101 ctgaccctgt gcgagtcgtc tccattgggg tcccggtgtc cgagttgctg gatgacccct
     8161 ctgggcctgc tggctccctg acttctgttg agttctgtgg gggaacgcac ctgcggaact
     8221 cgagtcatgc aggagctttt gtgatcgtga cggaagaagc cattgccaag ggtatccgga
     8281 ggattgtggc tgtcacaggt gccgaggccc agaaggccct caggaaagca gagagcttga
     8341 agaaatgtct ctctgtcatg gaagccaaag tgaaggctca gactgctcca aacaaggatg
     8401 tgcagaggga gatcgctgac cttggagagg ccctggccac tgcagtcatc ccccagtggc
     8461 agaaggatga attgcgggag actctcaaat ccctaaagaa ggtcatggat gacttggacc
     8521 gagccagcaa agccgatgtc cagaaacgag tgttagagaa gacgaagcag ttcatcgaca
     8581 gcaaccccaa ccagcctctt gtcatcctgg agatggagag cggcgcctca gccaaggccc
     8641 tgaatgaagc cttgaagctc ttcaagatgc actcccctca gacttctgcc atgctcttca
     8701 cggtggacaa tgaggctggc aagatcacgt gcctgtgtca agtcccccag aatgcagcca
     8761 atcggggctt aaaagccagc gagtgggtgc agcaggtgtc aggcttgatg gacggtaaag
     8821 gtggtggcaa ggatgtgtct gcacaggcca caggcaagaa cgttggctgc ctgcaggagg
     8881 cgctgcagct ggccacttcc ttcgcccagc tgcgcctcgg ggatgtaaag aactga
//
