LOCUS       sequence-375694-        6517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral delivery of guide RNA against mouse Runx1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6517)
  AUTHORS   Shin B, Hosokawa H, Romero-Wolf M, Zhou W, Masuhara K, Tobin VR, 
            Levanon D, Groner Y, Rothenberg EV
  TITLE     Runx1 and Runx3 drive progenitor to T-lineage transcriptome 
            conversion in mouse T cell commitment via dynamic genomic site 
            switching.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jan 26;118(4). pii: 2019655118. doi: 
            10.1073/pnas.2019655118.
  PUBMED    33479171
REFERENCE   2  (bases 1 to 6517)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2021-01-26"; volume: "118(4). pii: 
            2019655118. doi"; pages: " 10.1073/pnas.2019655118"
COMMENT     Sequence Label: E42_gRunx1_gRNA3_dTet_NGFR
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..60)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        45..374
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      225..360
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     396..415
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(496..513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(667..1255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(747..766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1426..2217,2218..2286))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2049..2068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2287..2391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2459..2477
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2577..2956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2957..3160
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3111..3131
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3161..3336
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3399..3598
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3799..4215
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4145..4167
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4183..4199
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        4231..4479
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4231..4251
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4402..4421
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        4619..4830
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4847..4856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5871..6296
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6399..6421)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cccagtcacg tagcgatagc ggagtgtaga tccggctgtg gaatgtgtgt cagttagggt
       61 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      121 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      181 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
      241 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
      301 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      361 taggcttttg caaaaagctt actggcttaa ctatgcggca tcagagcaga ttgtactgag
      421 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1081 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     1141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1741 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2041 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2401 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2461 taggcgtatc acgaggccct ttcgtcttca agaattagct tggccattgc atacgttgta
     2521 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca
     2581 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     2641 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2701 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2761 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2821 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2881 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2941 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3001 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3061 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3121 cggtaggcgt gtacggtggg aggtctatat aagcagagct caataaaaga gcccacaacc
     3181 cctcactcgg cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg tattcccaat
     3241 aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc
     3301 tgagtgattg actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag
     3361 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
     3421 tgtctgtccg attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta
     3481 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc
     3541 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga
     3601 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac
     3661 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg
     3721 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
     3781 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
     3841 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
     3901 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
     3961 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
     4021 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
     4081 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
     4141 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
     4201 ccttctctag gcgccggaat tgaaggatcc gagggcctat ttcccatgat tccttcatat
     4261 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     4321 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     4381 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     4441 ttcttggctt tatatatctt gtggaaagga cgaaacaccg agcggcgacc gcagcatggg
     4501 tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact
     4561 tgaaaaagtg gcaccgagtc ggtgcttttt ttctcgagtg gctccggtgc ccgtcagtgg
     4621 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     4681 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4741 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4801 tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcggtcgcca ccatggccac
     4861 aaccatggac gggccgcgcc tgctgctgtt gctgcttctg ggggtgtccc ttggaggtgc
     4921 caaggaggca tgccccacag gcctgtacac acacagcggt gagtgctgca aagcctgcaa
     4981 cctgggcgag ggtgtggccc agccttgtgg agccaaccag accgtgtgtg agccctgcct
     5041 ggacagcgtg acgttctccg acgtggtgag cgcgaccgag ccgtgcaagc cgtgcaccga
     5101 gtgcgtgggg ctccagagca tgtcggcgcc gtgcgtggag gccgacgacg ccgtgtgccg
     5161 ctgcgcctac ggctactacc aggatgagac gactgggcgc tgcgaggcgt gccgcgtgtg
     5221 cgaggcgggc tcgggcctcg tgttctcctg ccaggacaag cagaacaccg tgtgcgagga
     5281 gtgccccgac ggcacgtatt ccgacgaggc caaccacgtg gacccgtgcc tgccctgcac
     5341 cgtgtgcgag gacaccgagc gccagctccg cgagtgcaca cgctgggccg acgccgagtg
     5401 cgaggagatc cctggccgtt ggattacacg gtccacaccc ccagagggct cggacagcac
     5461 agcccccagc acccaggagc ctgaggcacc tccagaacaa gacctcatag ccagcacggt
     5521 ggcaggtgtg gtgaccacag tgatgggcag ctcccagccc gtggtgaccc gaggcaccac
     5581 cgacaacctc atccctgtct attgctccat cctggctgct gtggttgtgg gccttgtggc
     5641 ctacatagcc ttcaagaggt ggaacagtca tcgatatcct cgaggtcacc gcggtctaga
     5701 gtcgacctgc agccaagctt atcgataaat cgattagtcc aatttgttaa agacaggata
     5761 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     5821 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     5881 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     5941 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agggtcgacc
     6001 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     6061 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     6121 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     6181 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     6241 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg
     6301 ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa
     6361 gctggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6421 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6481 tcagcgggtg ttggcgggtg tcggggcgca gccatga
//
