LOCUS       PE2-VRER                9753 bp    DNA     circular SYN 22-MAY-2020
DEFINITION  Prime editing in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-PE2.
SOURCE      synthetic DNA construct.
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9753)
  TITLE     Liu Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9753)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 18, 2019 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|-13352647|
COMMENT     VNTDBDATE|-13352659|
COMMENT     LSOWNER|
COMMENT     VNTNAME|PE2-VRER|
COMMENT     VNTAUTHORNAME|Jiyeon Kweon|
COMMENT     VNTOAUTHORNAME|UNKNOWN|
COMMENT     Vector_NTI_Display_Data_(Do_Not_Edit!)
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COMMENT        #35=(CFSignal (CObList) "PE_1F" 274 0 0 2865 2883 0 (CStringList) 
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COMMENT                          "DNA 'PE2-VRER'" 1) 
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COMMENT                          "Prime editing in mammalian cells." 1) 
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COMMENT                          "Foreign object. Author: Jiyeon Kweon. Original author: UNKNOWN" 
COMMENT                          1) 
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COMMENT                          "Created: 05/22/20 02:22\277\300\310\304" 1) 
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COMMENT                          "length: 9753 bp" 1) 
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COMMENT                          "Original Source Database: GenBank" 1) 
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COMMENT                          "Division: SYN" 1) 
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COMMENT                          "Modification Date in the Original DB: 18-OCT-2019" 1) 
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COMMENT                          "Original Accession Number(s): <a href=\"ncbilink:dnarna/.\">.</a>" 
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COMMENT                          "Version in the Original DB: ." 1) 
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COMMENT                          "Sequence Source: synthetic DNA construct" 1) 
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COMMENT                          "Organizm(s): <a href=\"ncbilink:taxonomy/synthetic+DNA+construct\">synthetic DNA construct(synthetic DNA construct)</a>" 
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COMMENT                          "  JOURNAL   Exported Oct 18, 2019 from SnapGene Server 1.1.58" 
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COMMENT                                        "generates RNA-guided single strand nicks in DNA due to the H840A mutation in the HNH catalytic domain" 
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COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 626 (3 #13# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #140=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 626   End: 645 " 1) 
COMMENT                                     #141=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "T7 promoter, forward primer" 1) 
COMMENT                                     #142=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #143=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   626..645" 1))) "T7") 
COMMENT                             #144=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7057 (3 #19# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #145=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7057  End: 7074 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #146=(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                                     #147=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #148=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(7057..7074)" 1))) 
COMMENT                                   "BGH-rev") 
COMMENT                             #149=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7358 (3 #21# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #150=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7358  End: 7374 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #151=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "common sequencing primer, one of multiple similar variants" 
COMMENT                                           1) 
COMMENT                                     #152=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #153=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(7358..7374)" 1))) 
COMMENT                                   "M13 rev") 
COMMENT                             #154=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7358 (3 #22# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #155=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7358  End: 7374 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #156=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "In lacZ gene. Also called M13-rev" 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                                           "   complement(7358..7374)" 1))) 
COMMENT                                   "M13 Reverse") 
COMMENT                             #159=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7371 (3 #23# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #160=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7371  End: 7393 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #161=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "In lacZ gene" 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(7371..7393)" 1))) 
COMMENT                                   "M13/pUC Reverse") 
COMMENT                             #164=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7589 (3 #27# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #165=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7589  End: 7606 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #166=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "L4440 vector, forward primer" 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                                           "   complement(7589..7606)" 1))) 
COMMENT                                   "L4440") 
COMMENT                             #169=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7840 (3 #29# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #170=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7840  End: 7859 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #171=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "pBR322 origin, 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                                           "   complement(7840..7859)" 1))) 
COMMENT                                   "pBR322ori-F") 
COMMENT                             #174=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9142 (3 #31# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #175=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9142  End: 9161" 1) 
COMMENT                                     #176=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Ampicillin resistance gene, reverse primer" 
COMMENT                                           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                                           "   9142..9161" 1))) "Amp-R") 
COMMENT                             #179=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9563 (3 #33# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #180=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9563  End: 9582 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #181=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "pRS vectors, use to sequence yeast selectable marker" 
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(9563..9582)" 1))) 
COMMENT                                   "pRS-marker"))) 
COMMENT                           "Primer Binding Site (10 total)") 
COMMENT                     #184=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 30 (7 30 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #185=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 381 (3 #11# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #186=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 381   End: 584 " 1) 
COMMENT                                     #187=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "human cytomegalovirus (CMV) immediate early promoter" 
COMMENT                                           1) 
COMMENT                                     #188=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #189=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   381..584" 1))) "CMV promoter") 
COMMENT                             #190=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 626 (3 #14# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #191=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 626   End: 644 " 1) 
COMMENT                                     #192=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "promoter for bacteriophage T7 RNA polymerase" 
COMMENT                                           1) 
COMMENT                                     #193=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #194=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   626..644" 1))) "T7 promoter") 
COMMENT                             #195=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7406 (3 #25# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #196=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7406  End: 7436 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #197=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "promoter for the E. coli lac operon" 
COMMENT                                           1) 
COMMENT                                     #198=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #199=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(7406..7436)" 1))) 
COMMENT                                   "lac promoter") 
COMMENT                             #200=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 9380 (3 #32# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #201=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 9380  End: 9484 (Complementary)" 
COMMENT                                           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(9380..9484)" 1) 
COMMENT                                     #204=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #205=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /gene=\"bla\"" 1))) 
COMMENT                                   "AmpR promoter"))) 
COMMENT                           "Promoter Prokaryotic (4 total)") 
COMMENT                     #206=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 31 (7 31 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #207=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7382 (3 #24# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #208=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7382  End: 7398" 1) 
COMMENT                                     #209=(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                                     #210=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #211=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   7382..7398" 1) 
COMMENT                                     #212=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #213=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /bound_moiety=\"lac repressor encoded by lacI\"" 
COMMENT                                           1))) "lac operator") 
COMMENT                             #214=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7451 (3 #26# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #215=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7451  End: 7472" 1) 
COMMENT                                     #216=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "CAP binding activates transcription in the presence of cAMP." 
COMMENT                                           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                                           "   7451..7472" 1) 
COMMENT                                     #219=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #220=(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                     #221=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 33 (7 33 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #222=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 7760 (3 #28# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #223=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 7760  End: 8348 (Complementary)" 
COMMENT                                           1) 
COMMENT                                     #224=(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                                     #225=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Original Location Description:" 1) 
COMMENT                                     #226=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   complement(7760..8348)" 1) 
COMMENT                                     #227=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Qualifiers:" 1) 
COMMENT                                     #228=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "   /direction=LEFT" 1))) "ori"))) 
COMMENT                           "Replication Origin (1 total)") 
COMMENT                     #229=(CFolderPar 
COMMENT                           (CGroupPar (CParagraph 274 (7 274 0) 1 1 1 0 178) 
COMMENT                            (CObjectList 
COMMENT                             #230=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 2865 (3 #35# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #231=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 2865  End: 2883" 1))) "PE_1F") 
COMMENT                             #232=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 3229 (3 #34# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #233=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 3229  End: 3231" 1))) "H840A") 
COMMENT                             #234=(CFolderPar 
COMMENT                                   (CGroupPar 
COMMENT                                    (CParagraph 5235 (3 #36# 0) 1 2 2 0 194) 
COMMENT                                    (CObjectList 
COMMENT                                     #235=(CLinePar 
COMMENT                                           (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                           "Start: 5235  End: 5255 (Complementary)" 
COMMENT                                           1))) "PE_2R"))) "Overhang (3 total)"))) 
COMMENT                   "Feature Map") 
COMMENT              #236=(CFolderPar 
COMMENT                    (CGroupPar (CParagraph 12 (6 0) 1 1 0 0 178) 
COMMENT                     (CObjectList 
COMMENT                      #237=(CFolderPar 
COMMENT                            (CGroupPar (CParagraph 98 (0 0) 1 1 1 0 178) 
COMMENT                             (CObjectList 
COMMENT                              #238=(CFolderPar 
COMMENT                                    (CGroupPar (CParagraph 1 (0 0) 1 2 2 0 178) 
COMMENT                                     (CObjectList 
COMMENT                                      #239=(CLinePar 
COMMENT                                            (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                            "Start: 1     End: 9753" 1) 
COMMENT                                      #240=(CLinePar 
COMMENT                                            (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                            "Original Location Description:" 1) 
COMMENT                                      #241=(CLinePar 
COMMENT                                            (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                            "   1..9753" 1) 
COMMENT                                      #242=(CLinePar 
COMMENT                                            (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                            "Qualifiers:" 1) 
COMMENT                                      #243=(CLinePar 
COMMENT                                            (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                            "   /organism=\"synthetic DNA construct\"" 
COMMENT                                            1) 
COMMENT                                      #244=(CLinePar 
COMMENT                                            (CParagraph 0 (0 0) 1 2 3 0 180) 
COMMENT                                            "   /mol_type=\"other DNA\"" 1))) 
COMMENT                                    "Source_1"))) "Source (1 total)"))) 
COMMENT                    "Imported Features Not Shown on Map") 
COMMENT              #245=(CFolderPar 
COMMENT                    (CGroupPar (CParagraph 13 (0 0) 1 1 0 0 178) 
COMMENT                     (CObjectList 
COMMENT                      #246=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108909008 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #247=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #0# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #248=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #0# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 8014 " 1) 
COMMENT                                      #249=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #0# 2) 1 2 2 0 191) 
COMMENT                                            " N2: 9260 " 1))))) "ApaLI: 2 sites") 
COMMENT                            1 5 "GTGCAC" "CACGTG") 
COMMENT                      #250=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108909040 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #251=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #1# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #252=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #1# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 2978 " 1) 
COMMENT                                      #253=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #1# 2) 1 2 2 0 191) 
COMMENT                                            " N2: 4457 " 1) 
COMMENT                                      #254=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #1# 3) 1 2 2 0 191) 
COMMENT                                            " N3: 5634 " 1) 
COMMENT                                      #255=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #1# 4) 1 2 2 0 191) 
COMMENT                                            " N4: 5663 " 1))))) "AvaI: 4 sites") 
COMMENT                            1 5 "CYCGRG" "GRGCYC") 
COMMENT                      #256=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108908912 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #257=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #2# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #258=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #2# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 4835 " 1) 
COMMENT                                      #259=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #2# 2) 1 2 2 0 191) 
COMMENT                                            " N2: 5547 " 1))))) "BamHI: 2 sites") 
COMMENT                            1 5 "GGATCC" "CCTAGG") 
COMMENT                      #260=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108909104 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #261=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #3# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #262=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #3# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 9531 " 1))))) "ClaI: 1 site") 
COMMENT                            2 4 "ATCGAT" "TAGCTA") 
COMMENT                      #263=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108908496 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #264=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #4# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #265=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #4# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 6980 " 1))))) "EcoRI: 1 site") 
COMMENT                            1 5 "GAATTC" "CTTAAG") 
COMMENT                      #266=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108909200 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList)) "HindIII: 0 sites") 1 5 "AAGCTT" 
COMMENT                            "TTCGAA") 
COMMENT                      #267=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108908464 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #268=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #6# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #269=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #6# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 377 " 1) 
COMMENT                                      #270=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #6# 2) 1 2 2 0 191) 
COMMENT                                            " N2: 6195 " 1) 
COMMENT                                      #271=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #6# 3) 1 2 2 0 191) 
COMMENT                                            " N3: 6694 " 1))))) "NcoI: 3 sites") 
COMMENT                            1 5 "CCATGG" "GGTACC") 
COMMENT                      #272=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108908528 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList 
COMMENT                               #273=(CGroupPar 
COMMENT                                     (CParagraph 0 (10 #7# 0) 1 2 2 0 180) 
COMMENT                                     (CObjectList 
COMMENT                                      #274=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #7# 1) 1 2 2 0 191) 
COMMENT                                            " N1: 1506 " 1) 
COMMENT                                      #275=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #7# 2) 1 2 2 0 191) 
COMMENT                                            " N2: 2928 " 1) 
COMMENT                                      #276=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #7# 3) 1 2 2 0 191) 
COMMENT                                            " N3: 3132 " 1) 
COMMENT                                      #277=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #7# 4) 1 2 2 0 191) 
COMMENT                                            " N4: 3162 " 1) 
COMMENT                                      #278=(CLinePar 
COMMENT                                            (CParagraph 0 (1 #7# 5) 1 2 2 0 191) 
COMMENT                                            " N5: 4374 " 1))))) "PstI: 5 sites") 
COMMENT                            5 1 "CTGCAG" "GACGTC") 
COMMENT                      #279=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108908624 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList)) "SmaI: 0 sites") 3 3 "CCCGGG" 
COMMENT                            "GGGCCC") 
COMMENT                      #280=(CRSFolderPar 
COMMENT                            (CFolderPar 
COMMENT                             (CGroupPar (CParagraph 108909392 (8 0) 1 1 1 0 178) 
COMMENT                              (CObjectList)) "XmaI: 0 sites") 1 5 "CCCGGG" 
COMMENT                            "GGGCCC"))) "Restriction/Methylation Map")))) 
COMMENT       (CGraphView 
COMMENT        (CWStyleSheet 
COMMENT         (CObjectList 
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FEATURES             Location/Qualifiers
     source          1..9753
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /vntifkey="98"
     enhancer        1..380
                     /vntifkey="9"
                     /label=CMV\enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /vntifkey="30"
                     /label=CMV\promoter
                     /note="human cytomegalovirus (CMV) immediate early promoter"
     primer_bind     535..555
                     /vntifkey="28"
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /vntifkey="28"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /vntifkey="30"
                     /label=T7\promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             694..714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /vntifkey="4"
                     /label=SV40\NLS
     CDS             715..4815
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /vntifkey="4"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due to the H840A mutation in the HNH catalytic domain"
     CDS             6988..7008
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /vntifkey="4"
                     /label=SV40\NLS
     CDS             7017..7034
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /vntifkey="4"
                     /label=6xHis
     primer_bind     complement(7057..7074)
                     /vntifkey="28"
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse"
     polyA_signal    7063..7287
                     /vntifkey="25"
                     /label=bGH\poly(A)\signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7358..7374)
                     /vntifkey="28"
                     /label=M13\rev
                     /note="common sequencing primer, one of multiple similar variants"
     primer_bind     complement(7358..7374)
                     /vntifkey="28"
                     /label=M13\Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7371..7393)
                     /vntifkey="28"
                     /label=M13/pUC\Reverse
                     /note="In lacZ gene"
     protein_bind    7382..7398
                     /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(7406..7436)
                     /vntifkey="30"
                     /label=lac\promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7451..7472
                     /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(7589..7606)
                     /vntifkey="28"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7760..8348)
                     /direction=LEFT
                     /vntifkey="33"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     primer_bind     complement(7840..7859)
                     /vntifkey="28"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8519..9379)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /vntifkey="4"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
     primer_bind     9142..9161
                     /vntifkey="28"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9380..9484)
                     /gene="bla"
                     /vntifkey="30"
                     /label=AmpR\promoter
     primer_bind     complement(9563..9582)
                     /vntifkey="28"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable marker"
     overhang        3229..3231
                     /vntifkey="274"
                     /label=H840A
     overhang        2865..2883
                     /vntifkey="274"
                     /label=PE_1F
     overhang        complement(5235..5255)
                     /vntifkey="274"
                     /label=PE_2R
     misc_feature    3376..4794
                     /vntifkey="21"
                     /label=from\VRER
     misc_feature    3525..3525
                     /vntifkey="21"
                     /label=G\to\C\silent\mutation\in\original\vector
BASE COUNT     2616 a      2653 c      2576 g      1908 t 
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc 
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg 
      661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag 
      721 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac 
      781 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc 
      841 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg 
      901 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa 
      961 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag 
     1021 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg 
     1081 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg 
     1141 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag 
     1201 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag 
     1261 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc 
     1321 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa 
     1381 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc 
     1441 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa 
     1501 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc 
     1561 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc 
     1621 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag 
     1681 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg 
     1741 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt 
     1801 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg 
     1861 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg 
     1921 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg 
     1981 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc 
     2041 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc 
     2101 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac 
     2161 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc 
     2221 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag 
     2281 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag 
     2341 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag 
     2401 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg 
     2461 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac 
     2521 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg 
     2581 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc 
     2641 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc 
     2701 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg 
     2761 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc 
     2821 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac 
     2881 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg 
     2941 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg 
     3001 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg 
     3061 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac 
     3121 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac 
     3181 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct 
     3241 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac 
     3301 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg 
     3361 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc 
     3421 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa 
     3481 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac 
     3541 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg 
     3601 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac 
     3661 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag 
     3721 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc 
     3781 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg 
     3841 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc 
     3901 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg 
     3961 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc 
     4021 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag 
     4081 aaggactggg accctaagaa gtacggcggc ttcgtgagcc ccaccgtggc ctattctgtg 
     4141 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg 
     4201 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa 
     4261 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg 
     4321 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccagagaact gcagaaggga 
     4381 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag 
     4441 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag 
     4501 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc 
     4561 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga 
     4621 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc 
     4681 ttcaagtact ttgacaccac catcgaccgg aaggagtaca gaagcaccaa agaggtgctg 
     4741 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct 
     4801 cagctgggag gtgactctgg aggatctagc ggaggatcct ctggcagcga gacaccagga 
     4861 acaagcgagt cagcaacacc agagagcagt ggcggcagca gcggcggcag cagcacccta 
     4921 aatatagaag atgagtatcg gctacatgag acctcaaaag agccagatgt ttctctaggg 
     4981 tccacatggc tgtctgattt tcctcaggcc tgggcggaaa ccgggggcat gggactggca 
     5041 gttcgccaag ctcctctgat catacctctg aaagcaacct ctacccccgt gtccataaaa 
     5101 caatacccca tgtcacaaga agccagactg gggatcaagc cccacataca gagactgttg 
     5161 gaccagggaa tactggtacc ctgccagtcc ccctggaaca cgcccctgct acccgttaag 
     5221 aaaccaggga ctaatgatta taggcctgtc caggatctga gagaagtcaa caagcgggtg 
     5281 gaagacatcc accccaccgt gcccaaccct tacaacctct tgagcgggct cccaccgtcc 
     5341 caccagtggt acactgtgct tgatttaaag gatgcctttt tctgcctgag actccacccc 
     5401 accagtcagc ctctcttcgc ctttgagtgg agagatccag agatgggaat ctcaggacaa 
     5461 ttgacctgga ccagactccc acagggtttc aaaaacagtc ccaccctgtt taatgaggca 
     5521 ctgcacagag acctagcaga cttccggatc cagcacccag acttgatcct gctacagtac 
     5581 gtggatgact tactgctggc cgccacttct gagctagact gccaacaagg tactcgggcc 
     5641 ctgttacaaa ccctagggaa cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc 
     5701 cagaaacagg tcaagtatct ggggtatctt ctaaaagagg gtcagagatg gctgactgag 
     5761 gccagaaaag agactgtgat ggggcagcct actccgaaga cccctcgaca actaagggag 
     5821 ttcctaggga aggcaggctt ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc 
     5881 cccctgtacc ctctcaccaa accggggact ctgtttaatt ggggcccaga ccaacaaaag 
     5941 gcctatcaag aaatcaagca agctcttcta actgccccag ccctggggtt gccagatttg 
     6001 actaagccct ttgaactctt tgtcgacgag aagcagggct acgccaaagg tgtcctaacg 
     6061 caaaaactgg gaccttggcg tcggccggtg gcctacctgt ccaaaaagct agacccagta 
     6121 gcagctgggt ggcccccttg cctacggatg gtagcagcca ttgccgtact gacaaaggat 
     6181 gcaggcaagc taaccatggg acagccacta gtcattctgg ccccccatgc agtagaggca 
     6241 ctagtcaaac aaccccccga ccgctggctt tccaacgccc ggatgactca ctatcaggcc 
     6301 ttgcttttgg acacggaccg ggtccagttc ggaccggtgg tagccctgaa cccggctacg 
     6361 ctgctcccac tgcctgagga agggctgcaa cacaactgcc ttgatatcct ggccgaagcc 
     6421 cacggaaccc gacccgacct aacggaccag ccgctcccag acgccgacca cacctggtac 
     6481 acggatggaa gcagtctctt acaagaggga cagcgtaagg cgggagctgc ggtgaccacc 
     6541 gagaccgagg taatctgggc taaagccctg ccagccggga catccgctca gcgggctgaa 
     6601 ctgatagcac tcacccaggc cctaaagatg gcagaaggta agaagctaaa tgtttatact 
     6661 gatagccgtt atgcttttgc tactgcccat atccatggag aaatatacag aaggcgtggg 
     6721 tggctcacat cagaaggcaa agagatcaaa aataaagacg agatcttggc cctactaaaa 
     6781 gccctctttc tgcccaaaag acttagcata atccattgtc caggacatca aaagggacac 
     6841 agcgccgagg ctagaggcaa ccggatggct gaccaagcgg cccgaaaggc agccatcaca 
     6901 gagactccag acacctctac cctcctcata gaaaattcat caccctctgg cggctcaaaa 
     6961 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta accggtcatc 
     7021 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc 
     7081 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca 
     7141 ctgtcctttc ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta 
     7201 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc 
     7261 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga 
     7321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 
     7381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 
     7441 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 
     7501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 
     7561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 
     7621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 
     7681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 
     7741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 
     7801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 
     7861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 
     7921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 
     7981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 
     8041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 
     8101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 
     8161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 
     8221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 
     8281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 
     8341 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact 
     8401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 
     8461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 
     8521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 
     8581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 
     8641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 
     8701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 
     8761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 
     8821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 
     8881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 
     8941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 
     9001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 
     9061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 
     9121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 
     9181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 
     9241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 
     9301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 
     9361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 
     9421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 
     9481 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg 
     9541 atcccctagg gtctactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 
     9601 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 
     9661 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc 
     9721 tgcttcgcga tgtacgggcc agatatacgc gtt 
//
