LOCUS       sequence-375689-        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_gRNA1_dTet_NGFR
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..79)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        64..393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     415..434
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(515..532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(686..1274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(766..785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1445..2236,2237..2305))
                     /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     2068..2087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2306..2410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2478..2496
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2596..2975
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2976..3179
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3130..3150
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3180..3355
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3418..3617
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3818..4234
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4164..4186
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4202..4218
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        4250..4498
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4250..4270
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4421..4440
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        4638..4849
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4866..4875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5890..6315
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6418..6440)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg
       61 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      301 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
      361 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat
      421 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
      481 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1741 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2041 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     2101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2401 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     2461 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattagctt
     2521 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa
     2581 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     2641 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     2701 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     2761 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     2821 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     2881 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     2941 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     3001 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     3061 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     3121 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     3181 aataaaagag cccacaaccc ctcactcggc gcgccagtct tccgatagac tgcgtcgccc
     3241 gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gtctcgctgt
     3301 tccttgggag ggtctcctct gagtgattga ctacccacga cgggggtctt tcatttgggg
     3361 gctcgtccgg gatttggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
     3421 ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgtttg atgttatgcg
     3481 cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga
     3541 cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt
     3601 ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt
     3661 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
     3721 ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct
     3781 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
     3841 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
     3901 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
     3961 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
     4021 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
     4081 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
     4141 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
     4201 cctttatcca gccctcactc cttctctagg cgccggaatt gaaggatccg agggcctatt
     4261 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     4321 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     4381 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     4441 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
     4501 ctcgtgctgg catctacggt ttaagagcta tgctggaaac agcatagcaa gtttaaataa
     4561 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tctcgagtgg
     4621 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4681 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4741 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4801 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg
     4861 cggtcgccac catggccaca accatggacg ggccgcgcct gctgctgttg ctgcttctgg
     4921 gggtgtccct tggaggtgcc aaggaggcat gccccacagg cctgtacaca cacagcggtg
     4981 agtgctgcaa agcctgcaac ctgggcgagg gtgtggccca gccttgtgga gccaaccaga
     5041 ccgtgtgtga gccctgcctg gacagcgtga cgttctccga cgtggtgagc gcgaccgagc
     5101 cgtgcaagcc gtgcaccgag tgcgtggggc tccagagcat gtcggcgccg tgcgtggagg
     5161 ccgacgacgc cgtgtgccgc tgcgcctacg gctactacca ggatgagacg actgggcgct
     5221 gcgaggcgtg ccgcgtgtgc gaggcgggct cgggcctcgt gttctcctgc caggacaagc
     5281 agaacaccgt gtgcgaggag tgccccgacg gcacgtattc cgacgaggcc aaccacgtgg
     5341 acccgtgcct gccctgcacc gtgtgcgagg acaccgagcg ccagctccgc gagtgcacac
     5401 gctgggccga cgccgagtgc gaggagatcc ctggccgttg gattacacgg tccacacccc
     5461 cagagggctc ggacagcaca gcccccagca cccaggagcc tgaggcacct ccagaacaag
     5521 acctcatagc cagcacggtg gcaggtgtgg tgaccacagt gatgggcagc tcccagcccg
     5581 tggtgacccg aggcaccacc gacaacctca tccctgtcta ttgctccatc ctggctgctg
     5641 tggttgtggg ccttgtggcc tacatagcct tcaagaggtg gaacagtcat cgatatcctc
     5701 gaggtcaccg cggtctagag tcgacctgca gccaagctta tcgataaatc gattagtcca
     5761 atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca
     5821 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
     5881 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     5941 ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac
     6001 agatggaaca gggtcgaccc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     6061 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     6121 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     6181 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     6241 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6301 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
     6361 acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     6421 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6481 acaagcccgt cagggcgcgt cagcgggtgt tggcggg
//
