LOCUS pEN-L1-M2NFI-L2 3561 bp DNA circular 06-SEP-2022
DEFINITION .
ACCESSION urn.local...tw-dt4jj29
KEYWORDS .
SOURCE
ORGANISM .
COMMENT This file is created by Vector NTI
http://www.invitrogen.com/
VNTDATE|459275237|
VNTDBDATE|459275237|
LSOWNER|
VNTNAME|pEN-L1-M2NFI-L2,0|
VNTAUTHORNAME|Demo User|
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0") 21 (CShapeMapEntry 8 "Shape 17" 1 "Label
0") 43 (CShapeMapEntry 8 "Shape 20" 1 "Label
14") 50 (CShapeMapEntry 0 "Shape 32" 1
"Signal Label") 70 (CShapeMapEntry 0 "Shape 24" 1
"Label 7") 68 (CShapeMapEntry 0 "Shape 24" 1
"Label 5") 67 (CShapeMapEntry 0 "ORF" 0 "ORF
Label") 94 (CShapeMapEntry 8 "Shape 2" 1
"Label 4") 88 (CShapeMapEntry 9 "Shape 11"
1 "Label 0") 87 (CShapeMapEntry 8 "Shape 18"
1 "Label 0") 86 (CShapeMapEntry 8 "Shape 19"
1 "Label 0") 85 (CShapeMapEntry 8 "Shape 23"
1 "Label 10")) 62.4709 62.4709 39 39 0.1 -13986) 1 1 0 1 1
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#179=(CGroupWidget (CWidget 10 (6 0) 1 2 0 0 Nil -184 100)
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(CWidget 4 (7 4 0) 1 2 0 0 Nil -951 100)
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#183=(CGroupWidget
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#185=(CLabel
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#186=(CGroupWidget
(CWidget 22 (7 22 0) 1 2 0 0 Nil -1232 100)
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#188=(CLabel
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0 -10 1.9369 0.75235 #187#)) (CObjectList))
#189=(CGroupWidget
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48 0) 1 2 0 0 Nil -1164 100) (CObjectList
#190=(CWideArrow
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(CObjectList)) #195=(CGroupWidget
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86 0) 1 2 0 0 Nil -1302 100) (CObjectList
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#200=(CGroupWidget (CWidget 11 (8 0) 1 2 0 0 Nil
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#201=(CGroupWidget
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(CObjectList)) #213=(CGroupWidget (CWidget
14 (16 0) 1 2 0 0 Nil -1054 100) (CObjectList)
(CObjectList)) #214=(CGroupWidget
(CWidget 12 (0 0) 1 2 0 0 Nil -883 100)
(CObjectList) (CObjectList))) (CObjectList))) (CSeqView 10
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(CStringList) 268747808 40451132 (CObList))).
FEATURES Location/Qualifiers
attL1 1..100
/Color="#00cccc"
/created_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="attL1"
/Transferred_Similarity="98.00%"
/Primary_Match="98.00%; Adjusted-Interval-0:0/0; attL1"
misc_feature 48
/Original_Bases="C"
/label="C"
/note="Geneious type: Editing History Replacement"
misc_feature 99..104
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
promoter 101..157
/modified_by="mamee"
/label="minimal p35S"
misc_feature 160^161
/Original_Bases="TAGCT"
/label="TAGCT"
/note="Geneious type: Editing History Deletion"
NLS 166..210
/created_by="wacae"
/note="Derived using Geneious Prime 2021.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="NLS c2"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; c2 NLS
(NLS c2)"
/modified_by="mamee"
/label="c2 NLS"
CDS join(211..675,865..1116)
/modified_by="mamee"
/label="GFP"
primer_bind 675..701
/%GC="37.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="None"
/Tm="59.5"
/annotation_group="60528016190065009578268b-b03c-4b7f-affc
-8d790dca596b"
/created_by="primer3"
/Sequence="CGTAAGTTTCTGCTTCTACCTTTGATA"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L1-M2NFI-L2"
/label="P48"
intron 676..864
/label="PIV2 intron"
attL2 complement(1118..1217)
/created_by="mamee"
/Color="#00cccc"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="attL2"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL2"
misc_feature 1118^1119
/Original_Bases="AA"
/label="AA"
/note="Geneious type: Editing History Deletion"
primer complement(1255..1275)
/note="Derived using Geneious Prime 2021.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="pCDB-Cas9 - GWM13R"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13R
(pCDB-Cas9 - GWM13R)"
/label="GWM13R"
misc_feature 1328..1329
/Original_Bases="TA"
/label="TA"
/note="Geneious type: Editing History Replacement"
promoter 1336..1386
/created_by="chgai"
/X-Transferred_From="pCG034_pKan_Kan-CBE"
/X-Transferred_Similarity="100.00%"
/X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG034_pKan_Kan-CBE)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG040_ABE-Nuc-NLS_Kan
-reporter_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG043_Kan-ABE-Dest_co
ntrol_ccdB)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG046_ABE-LbCas12a-NL
S_Kan_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG005_pGGD-pKan-KanR-
E)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG006_pGGA-Kan_promot
er-B)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG035_pKan_Kan)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG039_ABE-N7-NLS_Kan-
reporter_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG041_ABE-SV40-NLS_Ka
n-reporter_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG054_Kan_CBE_control
)"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="KanR"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (KanR)"
/label="kan"
misc_feature 1387..2196
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="KanR"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; KanR"
/label="KanR"
/note="Geneious type: Bacterial Selection"
primer_bind 1412..1431
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="25.9"
/Tm="59.8"
/annotation_group="60528013819306002b5c6d64-0728-4c06-8cfc
-efda2f3ec144"
/created_by="primer3"
/Sequence="CGAGGCCGCGATTAAATTCC"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L1-M2NFI-L2"
/label="P44"
primer_bind 2046..2065
/%GC="60.0"
/Hairpin_Tm="40.0"
/Self_Dimer_Tm="None"
/Tm="60.0"
/annotation_group="605280125087410051649037-b408-4c4f-be4d
-4f730d4118c8"
/created_by="primer3"
/Sequence="CGCAGACCGATACCAGGATC"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L1-M2NFI-L2"
/label="P42"
rep_origin 2343..2931
/created_by="thjac"
/modified_by="mamee"
/note="Derived using Geneious Prime 2021.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC
origin of replication"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0;
ColE1/pMB1/pBR322/pUC Ori (high-copy-number
ColE1/pMB1/pBR322/pUC origin of replication)"
/label="ColE1/pMB1/pBR322/pUC Ori"
primer_bind 2831..2850
/#_Off-target_Sites=0
/Off-target_Template="pEN-L1-M2NFI-L2"
/Sequence="GGGGAAACGCCTGGTATCTT"
/Tm="59.7"
/%GC="55.0"
/Hairpin_Tm="37.3"
/Self_Dimer_Tm="None"
/modified_by="mamee"
/label="P45"
primer_bind 3439..3458
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="None"
/Tm="60.0"
/annotation_group="60528015029841002acd09af-7d18-471d-9bb2
-ca2b24d898ca"
/created_by="primer3"
/Sequence="CTTCCGACTGAGCCTTTCGT"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L1-M2NFI-L2"
/label="P46"
primer 3530..3550
/note="Derived using Geneious Prime 2021.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="pCDB-Cas9 - GWM13F"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13F
(pCDB-Cas9 - GWM13F)"
/label="GWM13F"
ORIGIN
1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
61 tgctttttta taatgccaac tttgtacaaa aaagcagggc ttagcaagac ccttcctcta
121 tataaggaag ttcatttcat ttggagagga cacgctggct ccatgcagcc ttctcttaaa
181 cgcatgaaaa tagagccatc ttctcaacct atggtgagca agggcgagga gctgttcacc
241 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
301 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
361 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
421 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
481 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
541 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
601 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
661 gtctatatca tggccgtaag tttctgcttc tacctttgat atatatataa taattatcat
721 taattagtag taatataata tttcaaatat ttttttcaaa ataaaagaat gtagtatata
781 gcaattgctt ttctgtagtt tataagtgtg tatattttaa tttataactt ttctaatata
841 tgaccaaaat ttgttgatgt gcaggacaag cagaagaacg gcatcaaggt gaacttcaag
901 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
961 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
1021 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
1081 gccgggatca ctctcggcat ggacgagctg tacaagtacc cagctttctt gtacaaagtt
1141 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
1201 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
1261 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
1321 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
1381 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
1441 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
1501 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
1561 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
1621 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
1681 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
1741 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
1801 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
1861 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
1921 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
1981 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
2041 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
2101 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
2161 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
2221 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
2281 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
2341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
2401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
2461 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
2521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
2581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
2641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
2701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
2761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
2821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
2881 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
2941 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
3001 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
3061 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
3121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
3181 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
3241 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
3301 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
3361 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
3421 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
3481 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
3541 gccagtctta agctcgggcc c
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