LOCUS       sequence-250012-        5012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer reporter construct containing the chicken EdnrB enhancer, 
            E2 driving Cerulean activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5012)
  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 5012)
  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-Cerulean
FEATURES             Location/Qualifiers
     source          1..5012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        590..735
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     928..947
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      999..1008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1005..1007,1008..1010,1011..1724)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1050..1071)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1311..1330)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1658..1679
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1895..2016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1906..1925)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1960..1979
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2264..2281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2435..3023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2515..2534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3194..3985,3986..4054))
                     /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     3817..3836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4055..4159)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4186..4641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4273..4292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4483..4504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4772..4820
                     /note="synthetic polyadenylation signal"
     misc_feature    4834..4925
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4874..4893
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 accgccacac taggactaag tcgcttgtgt cttgtcatta caatgcttcc ttttgtccga
       61 tggctgcaaa tgcttgacag tcattagaaa gtcatccaag gaaatacttc ctttccatgt
      121 acaagatgct tgtagtggat ttttaacact gagctttctc agtaactcag atagcattag
      181 caaaagaata gtttcttaca gcctgcctct gcccttaagg ttcttttata aagagaacag
      241 aagctctttg ttgttactcc ttcacacgtt attaacagag gtttaacaga aagaagaggg
      301 gctcgttata gtccgctgca atttgtcttt ttcagttcat tcggatctcc atcatgtcca
      361 accagaagac ggattaaccc agatccaggg tggctgcttc catactaaac agaaaatgaa
      421 ttttaaaaag tcagacagtg taatctctct ctgccaagaa ccaggaccgc tagcccgggc
      481 tcgagatctg cgatctaagt aagcttgctg cttcatcccc gtggcccgtt gctcgcgttt
      541 gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac
      601 cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg
      661 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg
      721 cagcgacccg cttaacagcg tcaacagcgt gccgcagatc ccaagcttgc ggccgcttaa
      781 gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      841 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagg
      961 ttcaatacag ctcttaagcg gccgcaagct agcttgccgc caccatggtg agcaagggcg
     1021 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1081 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1141 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1201 cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1261 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1321 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1381 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacg
     1441 ccatcagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1501 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1561 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1621 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1681 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgagcctct agactccctc
     1741 agcggcgcgc cactggtacg agaacggtgg agtacctttc gagcgccggt aagaagcgtc
     1801 gcctgactaa aggttcgggt gcagaccagt tccgacacgc gtaagatctc acgtgtctag
     1861 agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     1921 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1981 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2041 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2101 tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg agagccttca acccagtcag
     2161 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2221 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
     2281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3241 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3601 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     4201 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4261 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4321 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4381 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4441 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4501 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4561 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4621 aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt tgggaagggc
     4681 gatcggtgcg ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata
     4741 ttaaggtacg ggaggtactt ggagcggccg caataaaata tctttatttt cattacatct
     4801 gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac
     4861 gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt
     4921 tctctatcga taggtaccga gctcgagttc catggttgct tgtaggaccg ctcaagtgct
     4981 tgcagctcac aattaccctg ctggcagttg ta
//
