LOCUS       pCMV-PE-spRY-2A-       10569 bp    DNA     circular SYN 18-AUG-2020
DEFINITION  CMV and T7 promoter expression plasmid for human codon optimized SpRY(A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1 335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-T7-SpRY-P2A-EGFP (RTW4830).
SOURCE      synthetic DNA construct.
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Kleinstiver
  TITLE     Kleinstiver lab CRISPR plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8451)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 20, 2020 from SnapGene Server 1.1.58
            http://www.snapgene.com
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTDATE|-13376264|
COMMENT     VNTDBDATE|-13479121|
COMMENT     LSOWNER|
COMMENT     VNTNAME|pCMV-PE-spRY-2A-EGFP|
COMMENT     VNTAUTHORNAME|Jiyeon Kweon|
COMMENT     VNTOAUTHORNAME|UNKNOWN|
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COMMENT                          "CMV and T7 promoter expression plasmid for human codon optimized SpRY(A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1 335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP." 
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COMMENT                                           "   1485..1864" 1))) "CMV enhancer"))) 
COMMENT                           "Enhancer (1 total)") 
COMMENT                     #133=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 21 (7 21 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #134=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 2136 (3 #10# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #135=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 2136  End: 2145" 1) 
COMMENT                                     #136=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Unknown feature type:regulatory  vertebrate consensus sequence for strong initiation  of translation (Kozak, 1987)" 
COMMENT                                           1) 
COMMENT                                     #137=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #138=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   2136..2145" 1) 
COMMENT                                     #139=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #140=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /regulatory_class=\"other\"" 1))) 
COMMENT                                   "Misc_Feature_1") 
COMMENT                             #141=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 4659 (3 #32# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #142=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 4659  End: 4661" 1))) "H840A") 
COMMENT                             #143=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 6246 (3 #31# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #144=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 6246  End: 8425" 1) 
COMMENT                                     #145=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #146=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   6246..8438" 1))) "RT fragments"))) 
COMMENT                           "Misc. Feature (3 total)") 
COMMENT                     #147=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 25 (7 25 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #148=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9363 (3 #21# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #149=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9363  End: 9587" 1) 
COMMENT                                     #150=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "bovine growth hormone polyadenylation signal" 
COMMENT                                           1) 
COMMENT                                     #151=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #152=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   9387..9611" 1))) 
COMMENT                                   "bGH poly(A) signal"))) 
COMMENT                           "PolyA Signal (1 total)") 
COMMENT                     #153=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 28 (7 28 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #154=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 873 (3 #2# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #155=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 873   End: 892 " 1) 
COMMENT                                     #156=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Ampicillin resistance gene, reverse primer" 
COMMENT                                           1) 
COMMENT                                     #157=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #158=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   873..892" 1))) "Amp-R") 
COMMENT                             #159=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 1294 (3 #4# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #160=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 1294  End: 1313 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #161=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "pRS vectors, use to sequence yeast selectable marker" 
COMMENT                                           1) 
COMMENT                                     #162=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #163=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(1294..1313)" 1))) 
COMMENT                                   "pRS-marker") 
COMMENT                             #164=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 2019 (3 #7# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #165=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 2019  End: 2039" 1) 
COMMENT                                     #166=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Human CMV immediate early promoter, forward primer" 
COMMENT                                           1) 
COMMENT                                     #167=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #168=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   2019..2039" 1))) "CMV-F") 
COMMENT                             #169=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 2110 (3 #8# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #170=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 2110  End: 2129" 1) 
COMMENT                                     #171=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "T7 promoter, forward primer" 1) 
COMMENT                                     #172=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #173=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   2110..2129" 1))) "T7") 
COMMENT                             #174=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 8637 (3 #16# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #175=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 8637  End: 8658 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #176=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "EGFP, reverse primer" 1) 
COMMENT                                     #177=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #178=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(8661..8682)" 1))) 
COMMENT                                   "EGFP-N") 
COMMENT                             #179=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 8898 (3 #17# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #180=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 8898  End: 8917 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #181=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "For distinguishing EGFP variants, reverse primer" 
COMMENT                                           1) 
COMMENT                                     #182=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #183=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(8922..8941)" 1))) 
COMMENT                                   "EXFP-R") 
COMMENT                             #184=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9245 (3 #18# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #185=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9245  End: 9266" 1) 
COMMENT                                     #186=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "EGFP, forward primer" 1) 
COMMENT                                     #187=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #188=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   9269..9290" 1))) "EGFP-C") 
COMMENT                             #189=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9357 (3 #20# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #190=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9357  End: 9374 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #191=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Bovine growth hormone terminator, reverse primer. Also called BGH reverse" 
COMMENT                                           1) 
COMMENT                                     #192=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #193=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(9381..9398)" 1))) 
COMMENT                                   "BGH-rev") 
COMMENT                             #194=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9658 (3 #22# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #195=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9658  End: 9674 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #196=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "common sequencing primer, one of multiple similar variants" 
COMMENT                                           1) 
COMMENT                                     #197=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #198=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(9682..9698)" 1))) 
COMMENT                                   "M13 rev") 
COMMENT                             #199=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9658 (3 #23# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #200=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9658  End: 9674 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #201=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "In lacZ gene. Also called M13-rev" 1) 
COMMENT                                     #202=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #203=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(9682..9698)" 1))) 
COMMENT                                   "M13 Reverse") 
COMMENT                             #204=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9671 (3 #24# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #205=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9671  End: 9693 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #206=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "In lacZ gene" 1) 
COMMENT                                     #207=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #208=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(9695..9717)" 1))) 
COMMENT                                   "M13/pUC Reverse") 
COMMENT                             #209=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9889 (3 #28# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #210=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9889  End: 9906 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #211=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "L4440 vector, forward primer" 1) 
COMMENT                                     #212=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #213=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(9913..9930)" 1))) 
COMMENT                                   "L4440") 
COMMENT                             #214=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 10140 (3 #30# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #215=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 10140  End: 10159 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #216=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "pBR322 origin, forward primer" 1) 
COMMENT                                     #217=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #218=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(10164..10183)" 1))) 
COMMENT                                   "pBR322ori-F"))) 
COMMENT                           "Primer Binding Site (13 total)") 
COMMENT                     #219=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 30 (7 30 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #220=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 1111 (3 #3# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #221=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 1111  End: 1215 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #222=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #223=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(1111..1215)" 1) 
COMMENT                                     #224=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #225=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /gene=\"bla\"" 1))) 
COMMENT                                   "AmpR promoter") 
COMMENT                             #226=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 1865 (3 #6# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #227=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 1865  End: 2068" 1) 
COMMENT                                     #228=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "human cytomegalovirus (CMV) immediate early promoter" 
COMMENT                                           1) 
COMMENT                                     #229=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #230=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   1865..2068" 1))) "CMV promoter") 
COMMENT                             #231=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 2110 (3 #9# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #232=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 2110  End: 2128" 1) 
COMMENT                                     #233=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "promoter for bacteriophage T7 RNA polymerase" 
COMMENT                                           1) 
COMMENT                                     #234=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #235=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   2110..2128" 1))) "T7 promoter") 
COMMENT                             #236=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9706 (3 #26# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #237=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9706  End: 9736 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #238=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "promoter for the E. coli lac operon" 
COMMENT                                           1) 
COMMENT                                     #239=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #240=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(9730..9760)" 1))) 
COMMENT                                   "lac promoter"))) 
COMMENT                           "Promoter Prokaryotic (4 total)") 
COMMENT                     #241=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 31 (7 31 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #242=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9682 (3 #25# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #243=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9682  End: 9698" 1) 
COMMENT                                     #244=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "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)." 
COMMENT                                           1) 
COMMENT                                     #245=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #246=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   9706..9722" 1) 
COMMENT                                     #247=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #248=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /bound_moiety=\"lac repressor encoded by lacI\"" 
COMMENT                                           1))) "lac operator") 
COMMENT                             #249=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9751 (3 #27# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #250=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9751  End: 9772" 1) 
COMMENT                                     #251=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "CAP binding activates transcription in the presence of cAMP." 
COMMENT                                           1) 
COMMENT                                     #252=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #253=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   9775..9796" 1) 
COMMENT                                     #254=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #255=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /bound_moiety=\"E. coli catabolite activator protein\"" 
COMMENT                                           1))) "CAP binding site"))) 
COMMENT                           "Protein Binding Site (2 total)") 
COMMENT                     #256=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 33 (7 33 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #257=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 10060 (3 #29# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #258=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 10060  End: 79   (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #259=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" 
COMMENT                                           1) 
COMMENT                                     #260=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #261=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(10084..79)" 1) 
COMMENT                                     #262=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #263=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /direction=LEFT" 1))) "ori"))) 
COMMENT                           "Replication Origin (1 total)"))) "Feature Map") 
COMMENT              #264=(CFolderPar 
COMMENT                    (CGroupPar (CParagraph 13 (0 0) 1 1 0 0 178) 
COMMENT                     (CObjectList 
COMMENT                      #265=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108902800 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #266=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #0# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #267=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #0# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 2104 " 1))))) "NotI: 1 site") 
COMMENT                            2 6 "GCGGCCGC" "CGCCGGCG"))) 
COMMENT                    "Restriction/Methylation Map")))) 
COMMENT       (CGraphView 
COMMENT        (CWStyleSheet 
COMMENT         (CObjectList 
COMMENT          #268=(CWidgetStyle "RSite Label" 1 (LOGPEN 0 0 13408563) 1 0 1 
COMMENT                (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Georgia") 0.555556 0 1 5 
COMMENT                "@N (@S)" 0) 
COMMENT          #269=(CWidgetStyle "Signal Label" 1 (LOGPEN 0 0 0) 1 0 1 
COMMENT                (LOGFONT 0 0 0 0 700 0 0 0 0 3 2 1 34 "Arial") 0.666667 0 1 1 
COMMENT                "@N" 0) 
COMMENT          #270=(CWidgetStyle "Molecule Label 2" 0 0 1 
COMMENT                (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 18 "Georgia") 0.555556 0 1 16 
COMMENT                "@L bp" 0) 
COMMENT          #271=(CWidgetStyle "Molecule Label 1" 0 0 1 
COMMENT                (LOGFONT 0 0 0 0 400 0 0 0 0 3 2 1 34 "Verdana") 0.833333 0 1 1 
COMMENT                "@N" 0) 
COMMENT          #272=(CWidgetStyle "Shape 3" 1 (LOGPEN 0 0 3355545) 1 1 
COMMENT                (LOGBRUSH 0 6724095 0) 0 0 1 (LOGSHAPE 9 1 0.8 1.8 0)) 
COMMENT          #273=(CWidgetStyle "Shape 1" 1 (LOGPEN 0 0 6723840) 1 1 
COMMENT                (LOGBRUSH 0 10079334 0) 0 0 1 (LOGSHAPE 9 1 0.8 1.8 0)) 
COMMENT          #274=(CWidgetStyle "Axis" 1 (LOGPEN 0 0 10079436) 2 1 
COMMENT                (LOGBRUSH 0 13434879 0) 0 0 1 (LOGSHAPE 10 1 0 0 0)) 
COMMENT          #275=(CWidgetStyle "Line 2" 1 (LOGPEN 0 0 6723840) 8 0 0 0 1 
COMMENT                (LOGSHAPE 1 1.9 0 0 0)) 
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FEATURES             Location/Qualifiers
     CDS             complement(250..1110)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /vntifkey="4"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
     primer_bind     873..892
                     /vntifkey="28"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1111..1215)
                     /gene="bla"
                     /vntifkey="30"
                     /label=AmpR\promoter
     primer_bind     complement(1294..1313)
                     /vntifkey="28"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable marker"
     enhancer        1485..1864
                     /vntifkey="9"
                     /label=CMV\enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1865..2068
                     /vntifkey="30"
                     /label=CMV\promoter
                     /note="human cytomegalovirus (CMV) immediate early promoter"
     primer_bind     2019..2039
                     /vntifkey="28"
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2110..2129
                     /vntifkey="28"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2110..2128
                     /vntifkey="30"
                     /label=T7\promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2136..2145
                     /regulatory_class="other"
                     /vntifkey="21"
                     /label=Misc_Feature_1
                     /note="Unknown feature type:regulatory  vertebrate consensus sequence for strong initiation  of translation (Kozak, 1987)"
     CDS             2142..3860
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /vntifkey="4"
                     /label=Cas9(N)
     CDS             8418..8438
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /vntifkey="4"
                     /label=SV40\NLS
     CDS             8451..8516
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /vntifkey="4"
                     /label=3xFLAG
     CDS             8535..8591
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /vntifkey="4"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides."
     CDS             8592..9311
                     /codon_start=1
                     /product="enhanced GFP"
                     /vntifkey="4"
                     /label=EGFP
                     /note="mammalian codon-optimized"
     primer_bind     complement(8637..8658)
                     /vntifkey="28"
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8898..8917)
                     /vntifkey="28"
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9245..9266
                     /vntifkey="28"
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             9317..9334
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /vntifkey="4"
                     /label=6xHis
     primer_bind     complement(9357..9374)
                     /vntifkey="28"
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse"
     polyA_signal    9363..9587
                     /vntifkey="25"
                     /label=bGH\poly(A)\signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9658..9674)
                     /vntifkey="28"
                     /label=M13\rev
                     /note="common sequencing primer, one of multiple similar variants"
     primer_bind     complement(9658..9674)
                     /vntifkey="28"
                     /label=M13\Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9671..9693)
                     /vntifkey="28"
                     /label=M13/pUC\Reverse
                     /note="In lacZ gene"
     protein_bind    9682..9698
                     /bound_moiety="lac repressor encoded by lacI"
                     /vntifkey="31"
                     /label=lac\operator
                     /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(9706..9736)
                     /vntifkey="30"
                     /label=lac\promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9751..9772
                     /bound_moiety="E. coli catabolite activator protein"
                     /vntifkey="31"
                     /label=CAP\binding\site
                     /note="CAP binding activates transcription in the presence of cAMP."
     primer_bind     complement(9889..9906)
                     /vntifkey="28"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10060..79)
                     /direction=LEFT
                     /vntifkey="33"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     primer_bind     complement(10140..10159)
                     /vntifkey="28"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6246..8425
                     /vntifkey="21"
                     /label=RT\fragments
     misc_feature    4659..4661
                     /vntifkey="21"
                     /label=H840A
BASE COUNT     2818 a      2912 c      2810 g      2029 t 
ORIGIN
        1 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 
       61 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 
      121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 
      181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 
      241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 
      301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 
      361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 
      421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 
      481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 
      541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 
      601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 
      661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 
      721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 
      781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 
      841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 
      901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 
      961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 
     1021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 
     1081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 
     1141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 
     1201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga 
     1261 tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg ccgcatagtt 
     1321 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat 
     1381 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag 
     1441 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac 
     1501 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 
     1561 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 
     1621 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 
     1681 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 
     1741 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 
     1801 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 
     1861 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 
     1921 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 
     1981 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 
     2041 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc 
     2101 cgcggccgct aatacgactc actataggga gagccgccac catggacaag aagtacagca 
     2161 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg 
     2221 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc 
     2281 tgatcggagc cctgctgttc gacagcggcg aaacagccga gagaacccgg ctgaagagaa 
     2341 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca 
     2401 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg 
     2461 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg 
     2521 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg 
     2581 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc 
     2641 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc 
     2701 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg 
     2761 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg 
     2821 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg 
     2881 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga 
     2941 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg 
     3001 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga 
     3061 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg 
     3121 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt 
     3181 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag 
     3241 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg 
     3301 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca 
     3361 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg 
     3421 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc 
     3481 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca 
     3541 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt 
     3601 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg 
     3661 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag 
     3721 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg 
     3781 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg 
     3841 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt 
     3901 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc 
     3961 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg 
     4021 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag 
     4081 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga 
     4141 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg 
     4201 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg 
     4261 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc 
     4321 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc 
     4381 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga 
     4441 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg 
     4501 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc 
     4561 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg 
     4621 aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc 
     4681 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga 
     4741 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc 
     4801 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg 
     4861 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga 
     4921 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg 
     4981 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc 
     5041 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg 
     5101 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg 
     5161 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc 
     5221 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca 
     5281 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg 
     5341 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc 
     5401 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca 
     5461 aagagtctat cagacccaag aggaacagcg ataagctgat cgccagaaag aaggactggg 
     5521 accctaagaa gtacggcggc ttcctgtggc ccaccgtggc ctattctgtg ctggtggtgg 
     5581 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca 
     5641 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct 
     5701 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg 
     5761 aaaacggccg gaagagaatg ctggcctctg ccaagcagct gcagaaggga aacgaactgg 
     5821 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg 
     5881 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg 
     5941 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc 
     6001 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg 
     6061 agaatatcat ccacctgttt accctgacca gactgggagc ccctagagcc ttcaagtact 
     6121 ttgacaccac catcgacccc aagcagtaca gaagcaccaa agaggtgctg gacgccaccc 
     6181 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag 
     6241 gtgactctgg aggatctagc ggaggatcct ctggcagcga gacaccagga acaagcgagt 
     6301 cagcaacacc agagagcagt ggcggcagca gcggcggcag cagcacccta aatatagaag 
     6361 atgagtatcg gctacatgag acctcaaaag agccagatgt ttctctaggg tccacatggc 
     6421 tgtctgattt tcctcaggcc tgggcggaaa ccgggggcat gggactggca gttcgccaag 
     6481 ctcctctgat catacctctg aaagcaacct ctacccccgt gtccataaaa caatacccca 
     6541 tgtcacaaga agccagactg gggatcaagc cccacataca gagactgttg gaccagggaa 
     6601 tactggtacc ctgccagtcc ccctggaaca cgcccctgct acccgttaag aaaccaggga 
     6661 ctaatgatta taggcctgtc caggatctga gagaagtcaa caagcgggtg gaagacatcc 
     6721 accccaccgt gcccaaccct tacaacctct tgagcgggct cccaccgtcc caccagtggt 
     6781 acactgtgct tgatttaaag gatgcctttt tctgcctgag actccacccc accagtcagc 
     6841 ctctcttcgc ctttgagtgg agagatccag agatgggaat ctcaggacaa ttgacctgga 
     6901 ccagactccc acagggtttc aaaaacagtc ccaccctgtt taatgaggca ctgcacagag 
     6961 acctagcaga cttccggatc cagcacccag acttgatcct gctacagtac gtggatgact 
     7021 tactgctggc cgccacttct gagctagact gccaacaagg tactcgggcc ctgttacaaa 
     7081 ccctagggaa cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc cagaaacagg 
     7141 tcaagtatct ggggtatctt ctaaaagagg gtcagagatg gctgactgag gccagaaaag 
     7201 agactgtgat ggggcagcct actccgaaga cccctcgaca actaagggag ttcctaggga 
     7261 aggcaggctt ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc cccctgtacc 
     7321 ctctcaccaa accggggact ctgtttaatt ggggcccaga ccaacaaaag gcctatcaag 
     7381 aaatcaagca agctcttcta actgccccag ccctggggtt gccagatttg actaagccct 
     7441 ttgaactctt tgtcgacgag aagcagggct acgccaaagg tgtcctaacg caaaaactgg 
     7501 gaccttggcg tcggccggtg gcctacctgt ccaaaaagct agacccagta gcagctgggt 
     7561 ggcccccttg cctacggatg gtagcagcca ttgccgtact gacaaaggat gcaggcaagc 
     7621 taaccatggg acagccacta gtcattctgg ccccccatgc agtagaggca ctagtcaaac 
     7681 aaccccccga ccgctggctt tccaacgccc ggatgactca ctatcaggcc ttgcttttgg 
     7741 acacggaccg ggtccagttc ggaccggtgg tagccctgaa cccggctacg ctgctcccac 
     7801 tgcctgagga agggctgcaa cacaactgcc ttgatatcct ggccgaagcc cacggaaccc 
     7861 gacccgacct aacggaccag ccgctcccag acgccgacca cacctggtac acggatggaa 
     7921 gcagtctctt acaagaggga cagcgtaagg cgggagctgc ggtgaccacc gagaccgagg 
     7981 taatctgggc taaagccctg ccagccggga catccgctca gcgggctgaa ctgatagcac 
     8041 tcacccaggc cctaaagatg gcagaaggta agaagctaaa tgtttatact gatagccgtt 
     8101 atgcttttgc tactgcccat atccatggag aaatatacag aaggcgtggg tggctcacat 
     8161 cagaaggcaa agagatcaaa aataaagacg agatcttggc cctactaaaa gccctctttc 
     8221 tgcccaaaag acttagcata atccattgtc caggacatca aaagggacac agcgccgagg 
     8281 ctagaggcaa ccggatggct gaccaagcgg cccgaaaggc agccatcaca gagactccag 
     8341 acacctctac cctcctcata gaaaattcat caccctctgg cggctcaaaa agaaccgccg 
     8401 acggcagcga attcgagccc aagaagaaga ggaaagtctc gagcggaggc gactacaaag 
     8461 accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagggag 
     8521 gcggtggaag cggcgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga 
     8581 atcctggacc gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 
     8641 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 
     8701 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 
     8761 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 
     8821 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 
     8881 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 
     8941 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 
     9001 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 
     9061 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 
     9121 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 
     9181 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 
     9241 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 
     9301 tgtacaagtg accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc 
     9361 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 
     9421 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 
     9481 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 
     9541 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 
     9601 aagaaccagc tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc 
     9661 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 
     9721 aagcataaag tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt 
     9781 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 
     9841 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 
     9901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 
     9961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 
    10021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 
    10081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 
    10141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 
    10201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 
    10261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 
    10321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 
    10381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 
    10441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 
    10501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 
    10561 ctcttgatc 
//
