LOCUS pHnS_2comp_Donor;smFP-V5_KO;Gphn#1 5592 bp DNA circular UNA 11-JUL-2025
DEFINITION .
ACCESSION urn.local...15-jebled8
VERSION urn.local...15-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..1559
/standard_name="ORF1"
source 1560..1604
/chromosome=6
/db_xref="taxon:10090"
/map="6 29.15 cM"
/mol_type="mRNA"
/organism="Mus musculus"
/strain="C57BL/6"
CDS 1560..1604
/codon_start=1
/transl_table=1
/translation="GGGGSGGGGSGGGGS"
/standard_name="Linker"
CDS 1611..1652
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
misc_feature 1611..2846
/standard_name="smFP-V5"
CDS 1662..1703
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 1713..1754
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
misc_feature 2157..2162
/standard_name="gaagac>gaggat, mutation on BbsI site"
CDS 2286..2327
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2337..2378
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2397..2438
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2448..2489
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2703..2744
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2754..2795
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
CDS 2805..2846
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/standard_name="V5 tag"
primer_bind complement(2837..2858)
/standard_name="smV5_R"
CDS 2859..2872
/codon_start=1
/locus_tag="X"
/transl_table=1
/translation="*LAS"
/standard_name="X"
misc_feature 2882..2904
/locus_tag="DRSR2(2)"
/standard_name="DRSR2+PAM"
misc_feature 2952..3081
/standard_name="ITR"
rep_origin 3152..3607
regulatory 3633..3737
/note="Geneious type: promoter"
/gene="bla"
/standard_name="AmpR promoter"
CDS join(3738..3806,3807..4598)
/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 4769..5357
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 agcattgcca gggccatccc 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 tgttgccaga ctccatcatt tctcggtttg
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 catgtgctgt agatatcacc aagggtttca
1381 gagctatgca caagagtgca tagcaagttg aaataaggct agtccgttta caacttggag
1441 cttgctccaa gtggcacccg agtcgggtgc tttttttctc gagtactagg atccattagg
1501 cgccaattct gcagacaaat ggctctaggc gatcgtaatc acccgagtgg gctcgagcag
1561 gtggaggcgg ttcaggcgga ggtggctctg gcggtggcgg atcgggtcga ggaaaaccta
1621 taccgaaccc cctccttgga cttgatagta ctcagcagca gggtaagccc attccaaacc
1681 cacttctcgg tctggatagc acacagcaac aaggtaagcc tatccctaac cctctgttgg
1741 gcctggactc cacgggccaa ggtgtgagta agggcgaaga actgtttacc ggcgtggtcc
1801 caattctggt tgagctcgac ggtgacgtta acggacacaa gtttagtgtg cgcggagaag
1861 gagagggaga tgccaccaac ggaaaattga cgctgaagtt tatatgcacc acgggaaagc
1921 tgccggtgcc gtggccgacc ttggttacaa ccctgggtgg cggagtccag tgtttcagcc
1981 gttacccaga ccacatgaag cagcatgact tctttaagag tgctatgccg gagggatacg
2041 tgcaggaacg gacgatcagt ttcaaagatg acggtactta caagacgcga gccgaggtga
2101 agttcgaggg agacaccttg gtgaacagga tagaacttaa gggaattgat ttcaaggagg
2161 atggtaatat tctgggtcac aagctggaat acaactttaa cagccataac gtctacatta
2221 ctgccgacaa gcagaagaac ggcataaagg cgaatttcaa aattcggcac aacgtggaag
2281 gtggtggcaa gccgatcccg aatcccctgc tgggcttgga tagcacgcag caacagggaa
2341 agcccatccc taaccccttg ctgggactcg acagcaccgg aggccagcaa ggcggtggca
2401 aaccgatacc caatcccctg ctcggcctgg atagcaccca gcagcaaggc aagccaatcc
2461 ccaaccctct gctgggcctc gactcgacag gtggcgatgg aagtgttcaa ctggcggacc
2521 actatcagca gaatacgccg atcggagatg gccccgtgct gttgccggat aatcactatc
2581 tgtcgactca gagcgttctg agtaaggacc caaacgaaaa acgagatcac atggtgttgt
2641 tggagttcgt gaccgccgcc ggcattacgt tgggaatgga cgagctgtac aagggccaag
2701 gaggcaagcc cataccaaat cccttgctgg gcctcgacag cactcagcaa caaggaaagc
2761 caataccaaa tccactgttg ggcctcgaca gtacgcaaca gcagggcaag cccatcccga
2821 acccgttgct gggactggat agtacctaaa ccggtgctta gctagctagc taaacggatc
2881 cgcgatcgta atcacccgag tgggctagag ggggtagata agtagcatgg cgggttaatc
2941 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
3001 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
3061 gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
3121 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
3181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
3241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
3301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
3361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
3421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
3481 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
3541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
3601 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
3661 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
3721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
3781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
3841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
3901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
3961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
4021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
4081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
4141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
4201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
4261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
4321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
4381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
4441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
4501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
4561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
4621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
4681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
4741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
4801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
4861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
4921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
4981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
5041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
5101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
5161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
5221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
5281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
5341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
5401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
5461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
5521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
5581 ttcattaatg ca
//