LOCUS pEN-R2-8-L3 3409 bp DNA circular 06-SEP-2022
DEFINITION .
ACCESSION urn.local...132-dt4s1xu
KEYWORDS .
SOURCE
ORGANISM .
COMMENT This file is created by Vector NTI
http://www.invitrogen.com/
ORIGDB|GenBank
VNTDATE|459272238|
VNTDBDATE|459272238|
LSOWNER|
VNTNAME|pEN-R2-8-L3,0|
VNTAUTHORNAME|Demo User|
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FEATURES Location/Qualifiers
attR2 complement(1..126)
/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="attR2"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; attR2"
misc_feature 1^2
/Original_Bases="CATAGTGACTGCATATGTTGTGTTTTACAGTATTATGTAGT
CTGTTTTTTATGCAAAATCTAATTTAATATATTGATATTTATATCATTTTACGTTTCT
CGTTCAACTTTCTTGTACAAAGT"
/label="CATAGTGACTGCATATGTTGTGTTTTACAGTATTATGTAGTCTGTTTTTT
ATGCAAAATCTAATTTAATATATTGATATTTATATCATTTTACGTTTCTCGTTCAACT
TTCTTGTACAAAGT"
/note="Geneious type: Editing History Deletion"
misc_feature 126..127
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
terminator 128..894
/created_by="lapau"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="OCST"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; tOCS (OCST)"
/label="tOCS"
primer_bind 541..560
/#_Off-target_Sites=0
/Off-target_Template="pEN-R2-8-L3"
/Sequence="CACCGAGCGGCGAACTAATA"
/Tm="60.2"
/%GC="55.0"
/Hairpin_Tm="44.6"
/Self_Dimer_Tm="13.0"
/modified_by="mamee"
/label="P57"
misc_feature 895
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
attL3 complement(896..991)
/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="attL3"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL3"
primer complement(1049..1069)
/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"
promoter 1130..1180
/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 1181..1990
/modified_by="wacae"
/note="Derived using Geneious Prime 2021.2.2 '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 1206..1225
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="25.9"
/Tm="59.8"
/annotation_group="6053150055719800c6079120-b214-42bf-8b94
-b9f980048ef6"
/created_by="primer3"
/Sequence="CGAGGCCGCGATTAAATTCC"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-R2-8-L3"
/label="P44"
primer_bind 1840..1859
/%GC="60.0"
/Hairpin_Tm="40.0"
/Self_Dimer_Tm="None"
/Tm="60.0"
/annotation_group="6053149938814400ce5b4c6b-5072-419b-a932
-301187d7b92c"
/created_by="primer3"
/Sequence="CGCAGACCGATACCAGGATC"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-R2-8-L3"
/label="P42"
rep_origin 2137..2725
/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"
misc_feature 2284
/Original_Bases="C"
/label="C"
/note="Geneious type: Editing History Replacement"
misc_feature 2529..2530
/Original_Bases="AT"
/label="AT"
/note="Geneious type: Editing History Replacement"
primer_bind 2625..2644
/%GC="55.0"
/Hairpin_Tm="37.3"
/Self_Dimer_Tm="None"
/Tm="59.7"
/annotation_group="6053150118162700304d2321-1838-48d9-ac0b
-17444104f96e"
/created_by="primer3"
/Sequence="GGGGAAACGCCTGGTATCTT"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-R2-8-L3"
/label="P45"
primer_bind 3233..3252
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="None"
/Tm="60.0"
/annotation_group="60531501766424005c85fed7-f872-436c-a4d1
-1c7b760513f0"
/created_by="primer3"
/Sequence="CTTCCGACTGAGCCTTTCGT"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-R2-8-L3"
/label="P46"
primer 3324..3344
/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"
misc_feature 3408
/Original_Bases="A"
/label="A"
/note="Geneious type: Editing History Replacement"
ORIGIN
1 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
61 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt tcttgtacaa
121 agtgggactg ctttaatgag atatgcgaga cgcctatgat cgcatgatat ttgctttcaa
181 ttctgttgtg cacgttgtaa aaaacctgag catgtgtagc tcagatcctt accgccggtt
241 tcggttcatt ctaatgaata tatcacccgt tactatcgta tttttatgaa taatattctc
301 cgttcaattt actgattgta ccctactact tatatgtaca atattaaaat gaaaacaata
361 tattgtgctg aataggttta tagcgacatc tatgatagag cgccacaata acaaacaatt
421 gcgttttatt attacaaatc caattttaaa aaaagcggca gaaccggtca aacctaaaag
481 actgattaca taaatcttat tcaaatttca aaagtgcccc aggggctagt atctacgaca
541 caccgagcgg cgaactaata acgctcactg aagggaactc cggttccccg ccggcgcgca
601 tgggtgagat tccttgaagt tgagtattgg ccgtccgctc taccgaaagt tacgggcacc
661 attcaacccg gtccagcacg gcggccgggt aaccgacttg ctgccccgag aattatgcag
721 catttttttg gtgtatgtgg gccccaaatg aagtgcaggt caaaccttga cagtgacgac
781 aaatcgttgg gcgggtccag ggcgaatttt gcgacaacat gtcgaggctc agcaggacct
841 gcaggcatgc aagctagctt actagtgatg catattctat agtgtcacct aaatacaact
901 ttattataca aagttggcat tataaaaaag cattgcttat caatttgttg caacgaacag
961 gtcactatca gtcaaaataa aatcattatt tggagctcca tggtagcgtt aacgcggccg
1021 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
1081 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
1141 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
1201 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
1261 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
1321 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
1381 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
1441 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
1501 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
1561 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
1621 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
1681 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
1741 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
1801 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
1861 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
1921 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
1981 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
2041 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
2101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
2161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
2221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
2281 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
2341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
2401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
2461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
2521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
2581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
2641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
2701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
2761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
2821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
2881 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
2941 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
3001 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
3061 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
3121 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
3181 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
3241 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
3301 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag
3361 ctctagagct cgaattctac aggtcactaa taccatctaa gtagttggt
//