LOCUS pHnS_2comp_Donor;smFP-V5_KO;Template_ORF0 5593 bp DNA circular UNA 11-JUL-2025
DEFINITION .
ACCESSION urn.local...17-jebled8
VERSION urn.local...17-jebled8
KEYWORDS .
SOURCE
ORGANISM .
FEATURES Location/Qualifiers
misc_feature 1..130
/standard_name="ITR"
source 169..1501
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 172..190
/standard_name="Primer site #1"
misc_feature 191..480
/standard_name="bU6"
misc_feature 481..500
/standard_name="gRNA #1"
misc_feature 483..488
/standard_name="BbsI"
misc_feature 501..594
/standard_name="Scaffold cr1"
misc_feature 563..574
/standard_name="T-lock"
misc_feature 595..601
/standard_name="polyT"
misc_feature 602..621
/standard_name="Primer site #2"
regulatory 622..934
/note="Geneious type: promoter"
/note="RNA polymerase III promoter for mouse U6 snRNA (Das
et al., 1988)"
/standard_name="mU6 promoter"
primer_bind 853..873
/standard_name="mU6-F"
primer_bind complement(914..935)
/standard_name="SEQ_mU6_last_R"
misc_feature 936..955
/standard_name="gRNA #2"
misc_feature 956..1051
/standard_name="Scaffold cr2"
misc_feature 1018..1029
/standard_name="T-lock"
misc_feature 1052..1058
/standard_name="polyT"
misc_feature 1059..1077
/standard_name="Primer site #3"
misc_feature 1106..1354
/standard_name="hU6"
primer_bind 1106..1126
/standard_name="hU6-F"
primer_bind 1277..1296
/standard_name="LKO.1 5'"
misc_feature 1355..1374
/standard_name="gRNA #3"
misc_feature 1367..1372
/standard_name="BbsI"
misc_feature 1375..1391
/standard_name="Scaffold cr3"
misc_feature 1396..1470
/standard_name="Scaffold cr3"
misc_feature 1437..1448
/standard_name="T-lock"
misc_feature 1471..1477
/standard_name="polyT"
misc_feature 1478..1499
/standard_name="Primer site #4"
misc_feature 1529..1551
/locus_tag="DRSR2"
/standard_name="DRSR2"
misc_feature 1552..1560
/standard_name="ORF0"
source 1561..1605
/chromosome=6
/db_xref="taxon:10090"
/map="6 29.15 cM"
/mol_type="mRNA"
/organism="Mus musculus"
/strain="C57BL/6"
CDS 1561..1605
/codon_start=1
/transl_table=1
/translation="GGGGSGGGGSGGGGS"
/standard_name="Linker"
CDS 1612..1653
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
misc_feature 1612..2847
/standard_name="smFP-V5"
CDS 1663..1704
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 1714..1755
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
misc_feature 2158..2163
/standard_name="gaagac>gaggat, mutation on BbsI site"
CDS 2287..2328
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2338..2379
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2398..2439
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2449..2490
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2704..2745
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2755..2796
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2806..2847
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
primer_bind complement(2838..2859)
/standard_name="smV5_R"
CDS 2860..2873
/codon_start=1
/locus_tag="X"
/transl_table=1
/translation="*LAS"
/standard_name="X"
misc_feature 2883..2905
/locus_tag="DRSR2(2)"
/standard_name="DRSR2+PAM"
misc_feature 2953..3082
/standard_name="ITR"
rep_origin 3153..3608
regulatory 3634..3738
/note="Geneious type: promoter"
/gene="bla"
/standard_name="AmpR promoter"
CDS join(3739..3807,3808..4599)
/codon_start=1
/gene="bla"
/note="confers resistance to ampicillin, carbenicillin,
and related antibiotics"
/product="β-lactamase"
/transl_table=1
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
/standard_name="AmpR"
rep_origin 4770..5358
ORIGIN
1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctaccg gtaggtccct
181 cgacgttaac cgtgaccgag cttgtctgcc cgcgcagtcc actagacaga cgcgcagagg
241 tcgggcgggc ctagggccaa tttcccatgg ttcccttcat ttgcatatat gatgtaataa
301 ggttatggag actattaaac ttagccctaa tcaaactata tgatgataat gtgtgtggta
361 caaaaggtca taacttatta tatactttga aacttaaaaa agggttacag tttagtcacc
421 ataactgtaa aattttttct attcttagct ttatatagtt cttgagaggc catgtttatg
481 ctgtcttcat catggccgac gtttaagagc tatgctggaa acagcatagc aagtttaaat
541 aaggctagtc cgttatcaac ttggacttcg gtccaagtgg caccgagtcg gtgctttttt
601 tccagtctga ctatcacgat ggatccgacg cgccatctct aggcccgcgc cggccccctc
661 gcacagactt gtgggagaag ctcggctact cccctgcccc ggttaatttg catataatat
721 ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca gataatctgt tctttttaat
781 actagctaca ttttacatga taggcttgga tttctataag agatacaaat actaaattat
841 tattttaaaa aacagcacaa aaggaaactc accctaactg taaagtaatt gtgtgttttg
901 agactataaa tatcccttgg agaaaagcct tgttggttcg cattatccga accatgtttg
961 agagctatgc agaaagctgc atagcaagtt caaataaggc tagtccgtac acaacttggg
1021 cttcggccta agtggcagcc gagtcggctg ctttttttgc tcgaatctac actcagctat
1081 ggcgcgcccc aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca
1141 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
1201 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
1261 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
1321 ggctttatat atcttgtgga aagccagaaa catgtaagcg tcgcaagaag acgggtttca
1381 gagctatgca caagagtgca tagcaagttg aaataaggct agtccgttta caacttggag
1441 cttgctccaa gtggcacccg agtcgggtgc tttttttctc gagtactagg atccattagg
1501 cgccaattct gcagacaaat ggctctaggc gatcgtaatc acccgagtgg gctcgagcaa
1561 ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcgggtcg aggaaaacct
1621 ataccgaacc ccctccttgg acttgatagt actcagcagc agggtaagcc cattccaaac
1681 ccacttctcg gtctggatag cacacagcaa caaggtaagc ctatccctaa ccctctgttg
1741 ggcctggact ccacgggcca aggtgtgagt aagggcgaag aactgtttac cggcgtggtc
1801 ccaattctgg ttgagctcga cggtgacgtt aacggacaca agtttagtgt gcgcggagaa
1861 ggagagggag atgccaccaa cggaaaattg acgctgaagt ttatatgcac cacgggaaag
1921 ctgccggtgc cgtggccgac cttggttaca accctgggtg gcggagtcca gtgtttcagc
1981 cgttacccag accacatgaa gcagcatgac ttctttaaga gtgctatgcc ggagggatac
2041 gtgcaggaac ggacgatcag tttcaaagat gacggtactt acaagacgcg agccgaggtg
2101 aagttcgagg gagacacctt ggtgaacagg atagaactta agggaattga tttcaaggag
2161 gatggtaata ttctgggtca caagctggaa tacaacttta acagccataa cgtctacatt
2221 actgccgaca agcagaagaa cggcataaag gcgaatttca aaattcggca caacgtggaa
2281 ggtggtggca agccgatccc gaatcccctg ctgggcttgg atagcacgca gcaacaggga
2341 aagcccatcc ctaacccctt gctgggactc gacagcaccg gaggccagca aggcggtggc
2401 aaaccgatac ccaatcccct gctcggcctg gatagcaccc agcagcaagg caagccaatc
2461 cccaaccctc tgctgggcct cgactcgaca ggtggcgatg gaagtgttca actggcggac
2521 cactatcagc agaatacgcc gatcggagat ggccccgtgc tgttgccgga taatcactat
2581 ctgtcgactc agagcgttct gagtaaggac ccaaacgaaa aacgagatca catggtgttg
2641 ttggagttcg tgaccgccgc cggcattacg ttgggaatgg acgagctgta caagggccaa
2701 ggaggcaagc ccataccaaa tcccttgctg ggcctcgaca gcactcagca acaaggaaag
2761 ccaataccaa atccactgtt gggcctcgac agtacgcaac agcagggcaa gcccatcccg
2821 aacccgttgc tgggactgga tagtacctaa accggtgctt agctagctag ctaaacggat
2881 ccgcgatcgt aatcacccga gtgggctaga gggggtagat aagtagcatg gcgggttaat
2941 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
3001 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
3061 agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
3121 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
3181 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
3241 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
3301 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
3361 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
3421 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
3481 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
3541 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
3601 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
3661 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
3721 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
3781 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
3841 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
3901 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
3961 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
4021 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
4081 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
4141 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
4201 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
4261 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
4321 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
4381 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
4441 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
4501 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
4561 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
4621 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
4681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
4741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
4801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
4861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
4921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
4981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
5041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
5101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
5161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
5221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
5281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
5341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
5401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
5461 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
5521 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
5581 attcattaat gca
//