LOCUS pAAV_hSyn1_DIO_A 5463 bp DNA circular 14-FEB-2019 SOURCE ORGANISM COMMENT This file is created by Vector NTI http://www.invitrogen.com/ COMMENT ORIGDB|GenBank COMMENT VNTDATE|-12686103| COMMENT VNTDBDATE|-12686103| COMMENT LSOWNER| COMMENT VNTNAME|pAAV_hSyn1_DIO_Axon-GFP_WPRE_BGHpA| COMMENT VNTAUTHORNAME|Demo User| COMMENT Vector_NTI_Display_Data_(Do_Not_Edit!) COMMENT (SXF COMMENT (CGexDoc "pAAV_hSyn1_DIO_Axon-GFP_WPRE_BGHpA" 0 5463 COMMENT (CDBMol 0 0 1 3606 1 0 0 0 0 "" "" 0 0 0 0 COMMENT (CObList COMMENT #0=(CClstep COMMENT (CStringList "AarI" "Acc65I" "AflII" "AsiSI" "AvrII" "BaeI" "BclI" COMMENT "BfrBI" "BlpI" "BmgBI" "BsiWI" "BsmI" "BspMI" "BstBI" "BstEII" COMMENT "BstZ17I" "Bsu36I" "EcoICRI" "EcoNI" "FseI" "HpaI" "KpnI" "M.BaeI" COMMENT "M.BclI" "M.EcaI" "M.EcoAI" "M.HpaI" "M.KpnBI" "M.KpnI" "M.MunI" COMMENT "M.PshAI" "M.SacI" "M.SnaBI" "M.Tth111I" "M.XcmI" "MfeI" "NdeI" COMMENT "NruI" "NsiI" "PmeI" "Ppu10I" "PpuMI" "PshAI" "PsrI" "RleAI" COMMENT "SacI" "SanDI" "SapI" "SexAI" "SfiI" "SnaBI" "SpeI" "Sse8647I" COMMENT "StuI" "SwaI" "Tth111I" "XcmI") COMMENT (CStringList "AscI" "AgeI" "ApaBI" "BsaBI" "BsrGI" "BstAPI" "BstXI" COMMENT "M.EcoBI" "M.EcoR124II" "M.KpnAI" "NcoI") COMMENT (CStringList "EcoRI" "AccI" "AloI" "AlwNI" "ApaI" "AseI" "BbvCI" COMMENT "BglII" "BplI" "BsaI" "BspEI" "DraIII" "EcoRV" "FspAI" "HincII" COMMENT "HindIII" "M.AccI" "M.AloI" "M.ApaI" "M.BbvCIA" "M.BglII" COMMENT "M.BstVI" "M.DraIII" "M.EcoKI" "M.EcoR124I" "M.EcoRI" "M.EcoRV" COMMENT "M.HincII" "M.HindIII" "M.NmeSI" "M.PhiBssHII" "M.Rrh4273I" COMMENT "M.SsoI" "M.StyLTIII" "M.StySPI" "M.TfiI" "M.VspI" "MluI" "PacI" COMMENT "PciI" "PflMI" "PmlI" "PsiI" "PspOMI" "PvuI" "SalI" "ScaI" "SciI" COMMENT "Sse232I" "TfiI" "XhoI") 1 "pAAV_hSyn1_DIO_Axon-GFP_WPRE_BGHpA" 0 COMMENT "pAAV-hSyn1-DIO-SSFO-EYFP-P2A-nlsdTomato-WPRE-BGHpA" COMMENT "axon_EGFP_Insert_correct" 0 "AscI" 1 1 0 0 0 0 "EcoRI" 1 1 0 0 0 0 COMMENT "AscI" 0 1 1 0 0 0 0 0 "EcoRI" 0 1 1 0 0 0 0 0 "" "" 0 "" "" 0 "" 0 COMMENT "" 0 0 0 0 0 0 0 2 0 2 "" "" 1 0 1 "" "" 1 0 0 -1 -1 1 "AscI" COMMENT "EcoRI" 0 "" "" "" "" "cgcgccctattattacttgt" "" "")) (CObList) COMMENT (CObList COMMENT #1=(CFragment 3 1 0 121 0 "" 0 "" "" "" "" "" "" COMMENT "pAAV-hSyn1-DIO-SSFO-EYFP-P2A-nlsdTomato-WPRE-BGHpA" "" 0 0 1 4407 COMMENT 1859 802 3418 802 0 "" "" "" "" "" "" (CObList) COMMENT (CTerm 1 0 0 "EcoRI" "" 1 0 0 0 0 0 0 0 0 0) COMMENT (CTerm 1 1 0 "AscI" "" 1 0 0 0 0 0 0 0 0 0) 1 0) COMMENT #2=(CFragment 3 1 0 121 0 "" 8 "" "" "" "" "" "" COMMENT "axon_EGFP_Insert_correct" "" 0 0 1 1056 803 1858 3 1058 0 "" "" "" COMMENT "" "" "" (CObList) (CTerm 1 0 0 "AscI" "" 1 0 0 0 0 0 0 0 0 0) COMMENT (CTerm 1 1 0 "EcoRI" "" 1 0 0 0 0 0 0 0 0 0) 1 0)) COMMENT (CObList COMMENT #3=(CFragment 3 2 0 121 0 "" 0 "" "" "" "" "" "" COMMENT "pAAV-hSyn1-DIO-SSFO-EYFP-P2A-nlsdTomato-WPRE-BGHpA" "" 0 0 1 4407 COMMENT 0 0 3418 802 0 "" "" "" "" "" "" (CObList) COMMENT (CTerm 1 0 0 "EcoRI" "" 1 0 0 0 0 0 0 0 0 0) COMMENT (CTerm 1 1 0 "AscI" "" 1 0 0 0 0 0 0 0 0 0) 1 0) COMMENT #4=(CFragment 3 2 0 121 0 "" 0 "" "" "" "" "" "" COMMENT "axon_EGFP_Insert_correct" "" 0 0 1 0 0 0 0 0 0 "" "" "" "" "" "" COMMENT (CObList COMMENT #5=(CFragD 21 "2X stop" 9 14 1 COMMENT "Misc. Feature, Name: 2X stop, From 9 To 14 (Complementary)") COMMENT #6=(CFragD 21 "LoxP" 1024 1057 0 COMMENT "Misc. Feature, Name: LoxP, From 1024 To 1057")) COMMENT (CTerm 1 0 0 "AscI" "" 1 0 0 0 0 0 0 0 0 0) COMMENT (CTerm 1 1 0 "EcoRI" "" 1 0 0 0 0 0 0 0 0 0) 1 0)) -1 "") COMMENT (CDocSetData 1 0 0 0 0 0 "MAIN" 1 1 1 1 0 0 1 1 0 1 10 10 4294967295 50 0 COMMENT 1 0 (CHomObj 0 0 0 3 75) (CWordArray) (CWordArray) COMMENT (CStringList "ApaLI" "AvaI" "BamHI" "ClaI" "EcoRI" "HindIII" "NcoI" COMMENT "PstI" "SmaI" "XmaI") (CStringList "atg" "gtg") COMMENT (CStringList "taa" "tga" "tag") (CObList) 1 "{(0,1),2}" 0 0 "" 0 COMMENT 4294967295 0 0 0 0 0 0 0 "MAIN" 0 0 30 0 COMMENT (CProteinMotifSearchObject 70 20 1 1 1 1 0 0 1 0 0 0 0 0)) COMMENT (CMolPar 0 0 0 0 4294967295 1 5463 0 0 0 0 0 0 0 0) (CStringList) COMMENT (CStringList) (CObList) (COAPar 25 250 50 0 6 4 3 7) COMMENT (COAPar 25 250 50 0 6 4 3 7) (COAPar 25 250 50 0 6 4 3 7) (CObList) COMMENT (CObList COMMENT #7=(CFSignal (CObList) "hSynapsin promoter" 21 0 0 164 658 1 COMMENT (CStringList) (CStringList) 1 1 1 1 "164..658") COMMENT #8=(CFSignal (CObList) "hSyn1-seq-F" 21 0 0 581 600 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "581..600") COMMENT #9=(CFSignal (CObList) "BGH-pA" 21 0 0 2500 2703 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "4059..4262") COMMENT #10=(CFSignal (CObList) "BGH-R" 27 0 1 2614 2633 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "complement(4173..4192)") COMMENT #11=(CFSignal (CObList) "loxP" 21 0 0 767 800 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "767..800") COMMENT #12=(CFSignal (CObList) "lox2722" 21 0 0 683 716 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "683..716") COMMENT #13=(CFSignal (CObList) "WPRE-R(JT)" 27 0 1 1984 2005 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "complement(3543..3564)") COMMENT #14=(CFSignal (CObList) "WPRE" 21 0 0 1872 2480 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "3431..4039") COMMENT #15=(CFSignal (CObList) "pUC ori" 33 0 0 4791 5458 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "6350..7017") COMMENT #16=(CFSignal (CObList) "AmpR" 21 0 0 3783 4640 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "5342..6199") COMMENT #17=(CFSignal (CObList) "f1 Origin" 33 0 0 2958 3264 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "4517..4823") COMMENT #18=(CFSignal (CObList) "R-ITR" 21 0 0 2726 2866 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "4285..4425") COMMENT #19=(CFSignal (CObList) "L-ITR" 21 0 0 1 141 1 (CStringList) COMMENT (CStringList) 1 1 1 1 "1..141") COMMENT #20=(CFSignal (CObList) "LoxP" 21 0 0 1824 1857 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"cyan\"" COMMENT "/ApEinfo_revcolor=\"green\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "1024..1057") COMMENT #21=(CFSignal (CObList) "Lox2722" 21 0 0 1740 1773 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"cyan\"" COMMENT "/ApEinfo_revcolor=\"green\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "940..973") COMMENT #22=(CFSignal (CObList) "GAP43 Sequence" 21 0 1 1634 1690 1 COMMENT (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"#7eff74\"" COMMENT "/ApEinfo_revcolor=\"#7eff74\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "complement(834..890)") COMMENT #23=(CFSignal (CObList) "6xHisTag" 21 0 1 1610 1627 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"#7eff74\"" COMMENT "/ApEinfo_revcolor=\"#7eff74\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "complement(810..827)") COMMENT #24=(CFSignal (CObList) "PRSET Leader" 21 0 1 1535 1633 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"#7eff74\"" COMMENT "/ApEinfo_revcolor=\"#7eff74\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "complement(735..833)") COMMENT #25=(CFSignal (CObList) "2X stop" 21 0 1 809 814 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"#7eff74\"" COMMENT "/ApEinfo_revcolor=\"#7eff74\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "complement(9..14)") COMMENT #26=(CFSignal (CObList) "kozak" 21 0 1 1694 1699 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"#ff8000\"" COMMENT "/ApEinfo_revcolor=\"green\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "complement(894..899)") COMMENT #27=(CFSignal (CObList) "EGFP" 21 0 1 815 1534 1 (CStringList) COMMENT (CStringList "/ApEinfo_fwdcolor=\"cyan\"" COMMENT "/ApEinfo_revcolor=\"green\"" COMMENT "/ApEinfo_graphicformat=\"arrow_data {{0 1 2 0 0 -1} {} 0}" COMMENT "width 5 offset 0\"") 1 1 1 1 "complement(15..734)")) (CObList) COMMENT (CObList) (CObList) (CObList) (CObList) (CObList) COMMENT (CTextView 0 COMMENT #28=(CGroupPar (CParagraph 0 (0 0) 1 2 0 0 180) COMMENT (CObjectList COMMENT #29=(CRefLinePar COMMENT (CLinePar (CParagraph 0 (0 0) 0 2 0 0 233) COMMENT "pAAV_hSyn1_DIO_Axon-GFP_WPRE_BGHpA" 2) 5 "" 0 4) COMMENT #30=(CFolderPar COMMENT (CGroupPar (CParagraph 1 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #31=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "DNA 'pAAV_hSyn1_DIO_Axon-GFP_WPRE_BGHpA'" 1) COMMENT #32=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Local object" 1) COMMENT #33=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Created: 02/14/19 05:18PM" 1) COMMENT #34=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Last Modified: 02/14/19 05:18PM" 1) COMMENT #35=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "length: 5463 bp" 1) COMMENT #36=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "storage type: Basic" 1) COMMENT #37=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "form: Circular" 1))) "General Description") COMMENT #38=(CFolderPar COMMENT (CGroupPar (CParagraph 2 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #39=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Original Source Database: GenBank" 1) COMMENT #40=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "Modification Date in the Original DB: 14-FEB-2019" 1))) COMMENT "Standard Fields") COMMENT #41=(CRefLinePar COMMENT (CLinePar (CParagraph 0 (0 0) 0 2 0 0 233) "Comments" 2) 1 "" COMMENT 0 0) COMMENT #42=(CFolderPar COMMENT (CGroupPar (CParagraph 8 (0 0) 1 2 0 0 178) (CObjectList)) COMMENT "Annotations") COMMENT #43=(CFolderPar COMMENT (CGroupPar (CParagraph 11 (0 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #44=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "!!!Please note: this description contains molecules" COMMENT 1) COMMENT #45=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) COMMENT "which were possibly changed since the design process!!!" COMMENT 1) COMMENT #46=(CFolderPar COMMENT (CGroupPar (CParagraph 0 (0 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #47=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Result Molecule: pAAV_hSyn1_DIO_Axon-GFP_WPRE_BGHpA" COMMENT 1) COMMENT #48=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #49=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Recipient: pAAV-hSyn1-DIO-SSFO-EYFP-P2A-nlsdTomato-WPRE-BGHpA" COMMENT 1) COMMENT #50=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Left Terminus: EcoRI #1 (total 1)" 1) COMMENT #51=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Right Terminus: AscI #1 (total 1)" 1) COMMENT #52=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Full digestion" 1) COMMENT #53=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #54=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Donor: axon_EGFP_Insert_correct " 1) COMMENT #55=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Left Terminus: AscI #1 (total 1)" 1) COMMENT #56=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Flank Region: 0" 1) COMMENT #57=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Right Terminus: EcoRI #1 (total 1)" 1) COMMENT #58=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Flank Region: 0" 1) COMMENT #59=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Full digestion" 1) COMMENT #60=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #61=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Ligation: cohesive termini at both junctions" COMMENT 1) COMMENT #62=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Required orientation achieved" 1) COMMENT #63=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #64=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Preselection: dephosphorylation" 1) COMMENT #65=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #66=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Transformation system: Bacteria" 1) COMMENT #67=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Intra-chromosomal replication" 1) COMMENT #68=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #69=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Recommended method(s) for clone analysis:" 1) COMMENT #70=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " PCR" 1) COMMENT #71=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #72=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Recommended oligonucleotide for colony hybridization" COMMENT 1) COMMENT #73=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "of cloned fragment in resulting molecule:" 1) COMMENT #74=(COlyLinePar COMMENT (CLinePar COMMENT (CParagraph 0 COMMENT (17 COMMENT #75=(COligo "" "CGCGCCCTATTATTACTTGT" "" 0 COMMENT (CStringList) 0) 0) 1 2 2 0 181) COMMENT " CGCGCCCTATTATTACTTGT" 1)) COMMENT #76=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) "" 1) COMMENT #77=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Important Restriction Sites:" 1) COMMENT #78=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Cloned fragment may be cut by AscI and EcoRI" COMMENT 1) COMMENT #79=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Recombinant lacks sites of:" 1) COMMENT #80=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " AarI Acc65I AflII AsiSI AvrII BaeI" 1) COMMENT #81=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " BclI BfrBI BlpI BmgBI BsiWI BsmI" 1) COMMENT #82=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " BspMI BstBI BstEII BstZ17I Bsu36I EcoICRI" 1) COMMENT #83=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " EcoNI FseI HpaI KpnI M.BaeI M.BclI" 1) COMMENT #84=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " M.EcaI M.EcoAI M.HpaI M.KpnBI M.KpnI M.MunI" COMMENT 1) COMMENT #85=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " M.PshAI M.SacI M.SnaBI M.Tth111I M.XcmI MfeI" COMMENT 1) COMMENT #86=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " NdeI NruI NsiI PmeI Ppu10I PpuMI" 1) COMMENT #87=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " PshAI PsrI RleAI SacI SanDI SapI" 1) COMMENT #88=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " SexAI SfiI SnaBI SpeI Sse8647I StuI" 1) COMMENT #89=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " SwaI Tth111I XcmI" 1) COMMENT #90=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Unique sites inside cloned fragment:" 1) COMMENT #91=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " AscI AgeI ApaBI BsaBI BsrGI BstAPI BstXI M.EcoBI" COMMENT 1) COMMENT #92=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " M.EcoR124II M.KpnAI NcoI" 1) COMMENT #93=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT "Unique sites outside cloned fragment:" 1) COMMENT #94=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " EcoRI AccI AloI AlwNI ApaI AseI BbvCI BglII" COMMENT 1) COMMENT #95=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " BplI BsaI BspEI DraIII EcoRV FspAI HincII HindIII" COMMENT 1) COMMENT #96=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " M.AccI M.AloI M.ApaI M.BbvCIA M.BglII M.BstVI M.DraIII M.EcoKI" COMMENT 1) COMMENT #97=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " M.EcoR124I M.EcoRI M.EcoRV M.HincII M.HindIII M.NmeSI M.PhiBssHII M.Rrh4273I" COMMENT 1) COMMENT #98=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " M.SsoI M.StyLTIII M.StySPI M.TfiI M.VspI MluI PacI PciI" COMMENT 1) COMMENT #99=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " PflMI PmlI PsiI PspOMI PvuI SalI ScaI SciI" 1) COMMENT #100=(CLinePar (CParagraph 0 (0 0) 1 2 2 0 180) COMMENT " Sse232I TfiI XhoI" 1))) "Step #1"))) COMMENT "Design Description (*)") COMMENT #101=(CFolderPar COMMENT (CGroupPar (CParagraph 12 (6 0) 1 1 0 0 178) COMMENT (CObjectList COMMENT #102=(CFolderPar COMMENT (CGroupPar (CParagraph 21 (7 21 0) 1 1 1 0 178) COMMENT (CObjectList COMMENT #103=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 1 (3 #19# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #104=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 1 End: 141 " 1) COMMENT #105=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #106=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 1..141" 1))) "L-ITR") COMMENT #107=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 164 (3 #7# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #108=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 164 End: 658 " 1) COMMENT #109=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #110=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT " 164..658" 1))) COMMENT "hSynapsin promoter") COMMENT #111=(CFolderPar COMMENT (CGroupPar COMMENT (CParagraph 581 (3 #8# 0) 1 2 2 0 194) COMMENT (CObjectList COMMENT #112=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Start: 581 End: 600 " 1) COMMENT #113=(CLinePar COMMENT (CParagraph 0 (0 0) 1 2 3 0 180) COMMENT "Original Location Description:" 1) COMMENT #114=(CLinePar COMMENT 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1.17389 COMMENT #341#)) (CObjectList))) (CObjectList)) COMMENT #343=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -185 100) COMMENT (CObjectList) (CObjectList)) COMMENT #344=(CGroupWidget (CWidget 11 (0 0) 1 2 0 0 Nil 767 100) COMMENT (CObjectList) (CObjectList)) COMMENT #345=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil 2082602612 100) COMMENT (CObjectList) (CObjectList))) (CObjectList))) COMMENT (CSeqView 10 10 (CObjectList) (CObList) 1 (CObList)) (CObList) -1052689 COMMENT (CStringList) 4294111986 4294111986 (CObList))) FEATURES Location/Qualifiers misc_feature 164..658 /vntifkey="21" /label=hSynapsin\promoter misc_feature 581..600 /vntifkey="21" /label=hSyn1-seq-F misc_feature 2500..2703 /vntifkey="21" /label=BGH-pA primer complement(2614..2633) /vntifkey="27" /label=BGH-R misc_feature 767..800 /vntifkey="21" /label=loxP misc_feature 683..716 /vntifkey="21" /label=lox2722 primer complement(1984..2005) /vntifkey="27" /label=WPRE-R(JT) misc_feature 1872..2480 /vntifkey="21" /label=WPRE rep_origin 4791..5458 /vntifkey="33" /label=pUC\ori misc_feature 3783..4640 /vntifkey="21" /label=AmpR rep_origin 2958..3264 /vntifkey="33" /label=f1\Origin misc_feature 2726..2866 /vntifkey="21" /label=R-ITR misc_feature 1..141 /vntifkey="21" /label=L-ITR misc_feature 1824..1857 /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=LoxP misc_feature 1740..1773 /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=Lox2722 source complement(1535..1693) /dnas_title="pAAV-hSynapsin1-axon-GCaMP6f" /ApEinfo_label="source" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="98" source complement(1700..1733) /dnas_title="pAAV-hSynapsin1-axon-GCaMP6f" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="98" misc_feature complement(1634..1690) /ApEinfo_fwdcolor="#7eff74" /ApEinfo_revcolor="#7eff74" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=GAP43\Sequence /note="GAP43 Sequence" misc_feature complement(1610..1627) /ApEinfo_fwdcolor="#7eff74" /ApEinfo_revcolor="#7eff74" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=6xHisTag /note="6xHisTag" misc_feature complement(1535..1633) /ApEinfo_fwdcolor="#7eff74" /ApEinfo_revcolor="#7eff74" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=PRSET\Leader /note="PRSET Leader" misc_feature complement(809..814) /ApEinfo_fwdcolor="#7eff74" /ApEinfo_revcolor="#7eff74" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=2X\stop misc_feature complement(1694..1699) /ApEinfo_fwdcolor="#ff8000" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=kozak misc_feature complement(815..1534) /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /vntifkey="21" /label=EGFP BASE COUNT 1133 a 1490 c 1472 g 1368 t ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtttaat taagtgtcta gactgcagag 181 ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct 241 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc 301 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag 361 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca 421 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc 481 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag 541 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg 601 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaaac cggctagagg 661 atcctctaga gtcgactccg gaataacttc gtataggata ctttatacga agttatgcag 721 aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa cttcgtatag 781 catacattat acgaagttat ggcgcgccct attattactt gtacagctcg tccatgccga 841 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg 901 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc 961 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga 1021 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat 1081 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct 1141 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct 1201 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc 1261 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga 1321 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc 1381 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc 1441 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca 1501 ccccggtgaa cagctcctcg cccttgctca ccatggtggc gagatcctta tcgtcatcgt 1561 cgtacagatc ccgacccatt tgctgtccac cagtcatgct agccatacca tgatgatgat 1621 gatgatgaga accaatcttt tggtcctcat cattcttttc aacctgtttg gttcttctca 1681 tacagcacag catgccaccg gtggcgggtg gatccttgct agcagcttga attgctagca 1741 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt 1801 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa 1861 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact 1921 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 1981 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 2041 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 2101 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 2161 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 2221 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 2281 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc 2341 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 2401 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 2461 gcctccccgc atcgataccg agcgctgctc gagagatctc gactgtgcct tctagttgcc 2521 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca 2581 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta 2641 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc 2701 atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 2761 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 2821 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 2881 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3001 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3061 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 3121 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3241 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat 3301 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 3361 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 3421 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 3481 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 3541 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 3601 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 3661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 3721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 3781 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 3841 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 3901 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 3961 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 4021 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 4081 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 4141 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 4201 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 4261 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 4321 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 4381 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 4441 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 4501 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 4561 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 4621 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 4681 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 4741 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 4801 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 4861 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 4921 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 4981 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 5041 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 5101 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 5161 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 5221 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 5281 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 5341 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 5401 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 5461 tgt //