LOCUS       sequence-249971-        5007 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 5007)
  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 5007)
  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..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    39..87
                     /note="synthetic polyadenylation signal"
     misc_feature    101..192
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     141..160
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        869..1014
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1092..1224
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1207..1226
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1284..1994
                     /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(1429..1447)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1697..1716
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2169..2290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2180..2199)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2234..2253
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2538..2555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2709..3297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2789..2808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3468..4259,4260..4328))
                     /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     4091..4110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4329..4433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4460..4915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4547..4566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4757..4778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
       61 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
      121 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
      181 gaacatttct ctatcgatag gtaccgagct cgagttgtgc agttgcttgt aggaccgctc
      241 aagtgcttgc agctcacaat taccctgctg gcagttgtaa ccgccgcact aggactaagt
      301 cgcttgtgtc ttgtcattac aatgcttcct tttgtccgat ggctgcaaat gcttgacagt
      361 cattagaaag tcatccaagg aaatacttcc tttccatgta caagatgctt gtagtggatt
      421 tttaacactg agctttctca gtaactcaga tagcattagc aaaagaatag tttcttacag
      481 cctgcctctg cccttaaggt tcttttataa agagaacaga agctctttgt tgttactcct
      541 tcacacgtta ttaacagagg tttaacagaa agaagagggg cttgttatag tccgctgcaa
      601 tttgtctttt tcagttcatt cggatctcca tcatgtccaa ccagaagacg gattaaccca
      661 gatccagggt ggctgcttcc atactaaaca gaaaatgaat tttaaaaagt cagacagtgt
      721 aatctctctc tgccaagaac cacggtggct agcccgggct cgagatctgc gatctaagta
      781 agcttgctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga
      841 aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt
      901 ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat
      961 taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaacagcgt
     1021 caacagcgtg ccgcagatcc caagcttgcg gccgcttaag ctgcagaagt tggtcgtgag
     1081 gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     1141 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     1201 tccactttgc ctttctctcc acaggtgtcc actcccaggt tcaatacagc tcttaagcgg
     1261 ccgcaagcta gcttgccgcc accatggtga gcaagggcga ggaggataac atggccatca
     1321 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     1381 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     1441 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1501 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1561 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1621 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1681 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1741 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1801 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1861 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1921 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1981 agctgtacaa gtgagcctct agactccctc agcggcgcgc cgcccttgat caaaaggcga
     2041 gccattgatc accgtccgcc gtgcataaca gctgcacatc gactcctagg gagtcgtcgt
     2101 ctcttaccac aactaggaga tctcacgtgt ctagagtcgg ggcggccggc cgcttcgagc
     2161 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2221 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2281 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2341 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     2401 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     2461 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     2521 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2881 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3121 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3541 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3601 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3901 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4201 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4321 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4381 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4441 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4501 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4561 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4621 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4681 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4741 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4801 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4861 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     4921 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4981 cgccagccca agctaccatg ataagta
//
