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 (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/3366121""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/3366121"">Das
                     et al., 1988</a>)"
                     /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="<i>bla</i>"
                     /standard_name="AmpR promoter"
     CDS             join(3739..3807,3808..4599)
                     /codon_start=1
                     /gene="<i>bla</i>"
                     /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
//
