LOCUS       sequence-250009-        3494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chicken sox3 gRNA target site 1, cloned into the pU6 sgRNA (backbone
            #92359).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3494)
  AUTHORS   Ling ITC, Sauka-Spengler T
  TITLE     Early chromatin shaping predetermines multipotent vagal neural crest
            into neural, neuronal and mesenchymal lineages.
  JOURNAL   Nat Cell Biol. 2019 Dec;21(12):1504-1517. doi: 
            10.1038/s41556-019-0428-9. Epub 2019 Dec 2.
  PUBMED    31792380
REFERENCE   2  (bases 1 to 3494)
  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"; doi: 
            "10.1038/s41556-019-0428-9"; journalName: "Nat Cell Biol"; date: 
            "2019-12"; volume: "21"; issue: "12"; pages: "1504-1517"
COMMENT     Sequence Label: pU6-Sox3gRNA-T1
FEATURES             Location/Qualifiers
     source          1..3494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(94..113)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(95..113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(118..134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(118..134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      801..1389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1290..1309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1719..1746)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1838..1924)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2148..2153,2154..2170,2171..2176)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2224..3414
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(2576..2595)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcgcaaacca acccttgatc ggggaagaac agtatgtcga gctatttttt gacttactgg
       61 ggatcaagcc tgattgggag aaaataaaat atcccctata gtgagtcgta ttacatggtc
      121 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
      181 gataaaaata tatcatcatg cctcctctag aggtctcagg accgcggccg ctccttcagg
      241 cactgccgac gacagcccag gcggaggtcc tgagcgccgg cgccaaattt gcataaagag
      301 ctacccagaa gccctcgcgc gcggaaacgg gcaaaaaagg gcttctaata tggaaatatt
      361 acgccgaatc gcgtcacaaa ttggctaagc gggcctaaga gttaacaaga tgtgctatta
      421 agcggagcct tttggtggga agaaacggaa cagtcactgt gttctaaaag aacttgcagg
      481 atgagccttt aaataccgca gtctcgacgc tcttagtcgg ttcatgtagg tctgcgccgg
      541 tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact
      601 tgaaaaagtg gcaccgagtc ggtgcttttt tgttttccat atttttttcg cattgctttc
      661 cttggctttt acacccttac gcagcctcgc ccccttttcc cagggagacc ctcgagccac
      721 ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
      781 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
      841 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      901 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
      961 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1021 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1081 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1141 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1201 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1261 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1321 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1381 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1441 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     1501 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     1561 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     1621 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     1681 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     1741 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     1801 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     1861 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     1921 tttgatgcct ggcagttccc tactctcgcg ttctgcatta atgaatcggc caacgcgcgg
     1981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2101 cagaatcagg ggataacgca ggaaagaaca tgaattaatt ctcatgtttg acagcttatc
     2161 atcgattagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
     2221 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
     2281 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac
     2341 agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc
     2401 gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg
     2461 ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggattctc
     2521 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     2581 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     2641 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctcctta
     2701 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
     2761 ctaatgcagg agtcgcataa gggagagcgc cgacccatgc ccttgagagc cttcaaccca
     2821 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     2881 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     2941 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     3001 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     3061 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     3121 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     3181 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     3241 ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat
     3301 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt
     3361 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa
     3421 gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc
     3481 ggagaactgt gaat
//
