LOCUS       sequence-250011-        3494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chicken sox3 gRNA target site 2, 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-T2
FEATURES             Location/Qualifiers
     source          1..3494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(279..306)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(398..484)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(708..713,714..730,731..736)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             784..1974
                     /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(1136..1155)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2148..2167)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2149..2167)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2172..2188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2172..2188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      2855..3443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3344..3363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
       61 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      121 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      181 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      241 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
      301 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
      361 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
      421 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
      481 tttgatgcct ggcagttccc tactctcgcg ttctgcatta atgaatcggc caacgcgcgg
      541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      661 cagaatcagg ggataacgca ggaaagaaca tgaattaatt ctcatgtttg acagcttatc
      721 atcgattagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
      781 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
      841 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac
      901 agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc
      961 gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg
     1021 ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggattctc
     1081 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     1141 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     1201 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctcctta
     1261 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
     1321 ctaatgcagg agtcgcataa gggagagcgc cgacccatgc ccttgagagc cttcaaccca
     1381 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     1441 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     1501 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     1561 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     1621 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     1681 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     1741 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     1801 ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat
     1861 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt
     1921 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa
     1981 gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc
     2041 ggagaactgt gaatgcgcaa accaaccctt gatcggggaa gaacagtatg tcgagctatt
     2101 ttttgactta ctggggatca agcctgattg ggagaaaata aaatatcccc tatagtgagt
     2161 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     2221 gttacattgc acaagataaa aatatatcat catgcctcct ctagaggtct caggaccgcg
     2281 gccgctcctt caggcactgc cgacgacagc ccaggcggag gtcctgagcg ccggcgccaa
     2341 atttgcataa agagctaccc agaagccctc gcgcgcggaa acgggcaaaa aagggcttct
     2401 aatatggaaa tattacgccg aatcgcgtca caaattggct aagcgggcct aagagttaac
     2461 aagatgtgct attaagcgga gccttttggt gggaagaaac ggaacagtca ctgtgttcta
     2521 aaagaacttg caggatgagc ctttaaatac cgcagtctcg acgctcttag tcgtctgcgc
     2581 cgtggacatc atggtttaag agctatgctg gaaacagcat agcaagttta aataaggcta
     2641 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt ccatattttt
     2701 ttcgcattgc tttccttggc ttttacaccc ttacgcagcc tcgccccctt ttcccaggga
     2761 gaccctcgag ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3001 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3481 tttgctggcc tttt
//
