LOCUS       sequence-250017-        5703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified pTK plasmid containing chicken EdnrB E2 enhancer driving 
            expression of full length Chicken Sox10 in frame with a Flag-tag and
            Avi-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5703)
  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 5703)
  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: pTK-EdnrbE2-Sox10-FLAG-tev-AVI
FEATURES             Location/Qualifiers
     source          1..5703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        130..275
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          353..485
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     468..487
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1940..1963
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2012..2056
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    2129..2250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2140..2159)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2194..2213
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2498..2515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2669..3257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2749..2768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3428..4219,4220..4288))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4051..4070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4289..4393)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4420..4875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4507..4526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4717..4738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5006..5054
                     /note="synthetic polyadenylation signal"
     misc_feature    5068..5159
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5108..5127
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 acctgttggc tagcccgggc tcgagatctg cgatctaagt aagcttgctg cttcatcccc
       61 gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt
      121 agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag
      181 atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc
      241 tcgaacaccg agcgaccctg cagcgacccg cttaacagcg tcaacagcgt gccgcagatc
      301 ccaagcttgc ggccgcttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat
      361 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
      421 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
      481 cacaggtgtc cactcccagg ttcaatacag ctcttaagcg gccgcaagct agcttgccgc
      541 caccatgatg gctgatgacc aagatctgtc ggaggtggag atgagcccag tgggttctga
      601 agaccaccac tgcctctcgc cgggaccatc catggcatcg gacaactctt cgcacctcgc
      661 cagctctggg aatggagaga tgggcaaggt caagaaggag cagcaagact cggaggcgga
      721 tgacgataag ttccctgtct gcatccgtga agcggtcagc caggtgctga gcggctatga
      781 ctggaccctg gtacccatgc ctgttcgggt caatggaagt aacaagagca aaccccacgt
      841 caagaggccc atgaacgcct tcatggtctg ggcgcaggct gcccggagga aattggctga
      901 ccagtacccg cacctccaca atgctgagct cagtaagacc ttggggaagc tctggaggct
      961 gctgaatgaa agtgataagc gtccctttat tgaagaggca gagcggctga ggatgcagca
     1021 caagaaggac caccccgact acaagtacca gccacggagg cggaaaaatg gcaaggccac
     1081 gcagggcgag ggcgagggcc aggtggaggg ggaggctggc ggggctgcct ctatccaggc
     1141 tcactacaag aatgcccacc tggaccacag gcatcctggg gaagggtcac ccatgtccga
     1201 tgggcacccg gagcactctt caggtcagag ccacggccct cccactcctc ccaccacccc
     1261 taagactgag ctgcaggcag gcaaagctga ctccaagcga gaagggcgtt cccttgggga
     1321 agggggaaag ccacacattg actttggcaa tgtggacatc ggggagatca gccatgaagt
     1381 gatgtccaac atggagacct ttgacgtcaa cgaatttgac caatacctgc cacccaacgg
     1441 acacgccggc cacccaggcc atgttggggg ctacgccgcg gccgctggct atggccttgg
     1501 gagcgccctg gctgcggcca gtggacactc agcctggatc tccaagcagc atggagtctc
     1561 cttgtccgcc accacgtcac cggtggtgga ctccaaagcc caggtgaaaa cagaggggtc
     1621 tgcccctgga ggtcattaca ccgatcagcc ctccacctcc cagatagctt acacatccct
     1681 gagtctgccc cactatggtt cggccttccc atccatctcc aggcctcagt ttgactatcc
     1741 agaccaccag ccctcgggac cctactacag ccactccagc caagcctctg gcctctactc
     1801 tgccttctcc tatatgggac cttcccagcg tcctctttac actgccatct ctgaccccgc
     1861 accctccgtg ccacagtccc atagtcccac acattgggaa cagcctgtgt atacgactct
     1921 ctccagacca gtaggaggtg actacaagga cgacgacgac aagggtggag cagctggagg
     1981 tgagaacctg tacttccagt tcatggctgg tggcctgaat gacatctttg aggcccagaa
     2041 gatcgagtgg catgaggact agtaatctgt acaagtaagc ggccgcgact ctagagtcgg
     2101 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2161 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2221 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2281 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2341 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     2401 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     2461 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3421 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3481 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3541 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3601 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3661 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3721 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3781 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3841 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3901 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3961 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4021 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4081 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4141 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4201 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4261 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4321 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4381 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4741 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4801 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4861 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4921 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     4981 tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5041 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
     5101 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta
     5161 tcgataggta ccgagctcga gttgccagtg cttgtaggac cgctcaagtg cttgcagctc
     5221 acaattaccc tgctggcagt tgtaaccgcc gcactaggac taagtcgctt gtgtcttgtc
     5281 attacaatgc ttccttttgt ccgatggctg caaatgcttg acagtcatta gaaagtcatc
     5341 caaggaaata cttcctttcc atgtacaaga tgcttgtagt ggatttttaa cactgagctt
     5401 tctcagtaac tcagatagca ttagcaaaag aatagtttct tacagcctgc ctctgccctt
     5461 aaggttcttt tataaagaga acagaagctc tttgttgtta ctccttcaca cgttattaac
     5521 agaggtttaa cagaaagaag aggggcttgt tatagtccgc tgcaatttgt ctttttcagt
     5581 tcattcggat ctccatcatg tccaaccaga agacggatta acccagatcc agggtggctg
     5641 cttccatact aaacagaaaa tgaattttaa aaagtcagac agtgtaatct ctctctgcca
     5701 aga
//
