LOCUS       sequence-244931-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer reporter construct containing the chicken EdnrB enhancer E2
            driving mCherry activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        821..966
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1044..1176
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1159..1178
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1230..1239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1236..1946
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1381..1399)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1649..1668
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1997..2118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2008..2027)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2062..2081
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2366..2383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2617..2636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3296..4087,4088..4156))
                     /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     3919..3938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4157..4261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4288..4743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4375..4394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4585..4606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttgt
      181 tgcttgtagg accgctcaag tgcttgcagc tcacaattac cctgctggca gttgtaaccg
      241 ccgcactagg actaagtcgc ttgtgtcttg tcattacaat gcttcctttt gtccgatggc
      301 tgcaaatgct tgacagtcat tagaaagtca tccaaggaaa tacttccttt ccatgtacaa
      361 gatgcttgta gtggattttt aacactgaac tttctcagta actcagatag cattagcaaa
      421 agaatagttt cttacagcct gcctctgccc ttaaggttct tttataaaga gaacagaagc
      481 tctttgttgt tactccttca cacgttatta acagaggttt aacagaaaga agaggggctt
      541 gttatagtcc gctgcaattt gtctttttca gttcattcgg atctccatca tgtccaacca
      601 gaagacggat taacccagat ccagggtggc tgcttccata ctaaacagaa aatgaatttt
      661 aaaaagtcag acagtgtaat ctctctctgc caagaaccgg ctagcccggg ctcgagatct
      721 gcgatctaag taagcttgct gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt
      781 gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa ccccgcccag
      841 cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc
      901 cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc
      961 gcttaacagc gtcaacagcg tgccgcagat cccaagcttg cggccgctta agctgcagaa
     1021 gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc
     1081 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     1141 gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag gttcaataca
     1201 gctcttaagc ggccgcaagc tagcttgccg ccaccatggt gagcaagggc gaggaggata
     1261 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1321 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1381 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1441 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1501 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1561 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1621 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1681 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     1741 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     1801 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     1861 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     1921 gcggcatgga cgagctgtac aagtaagcgg ccgcgactct agagtcgggg cggccggccg
     1981 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2041 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2101 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2161 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     2221 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     2281 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     2341 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4261 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     4321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4501 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4621 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4681 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4741 tttgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4801 cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac
     4861 ttggagcggc cgc
//
