LOCUS       sequence-250025-        5385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer reporter construct containing the chicken EdnrB enhancer E1
            driving Citrine activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5385)
  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 5385)
  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-EdnrbE1-Citrine
FEATURES             Location/Qualifiers
     source          1..5385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..65
                     /note="synthetic polyadenylation signal"
     misc_feature    79..170
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     119..138
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1222..1367
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1445..1577
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1560..1579
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1631..1640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1637..1639,1640..1642,1643..2353)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1682..1703)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1943..1962)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2290..2311
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2525..2646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2536..2555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2590..2609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2894..2911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3065..3653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3145..3164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3824..4615,4616..4684))
                     /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     4447..4466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4685..4789)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4816..5271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4903..4922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5113..5134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
       61 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
      121 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt
      181 accgagctcg agttgccagg caacacaagg aaggtcctgc tttctggttg tccaactaaa
      241 gctacccaga tttagcttgc tggctcttcc tttccctttt caatatctaa attgcattac
      301 aagatgataa gaaatacact gaatactccc acaatggttg ctacaggaat gtgatgtagt
      361 tacagatgag aagagagatg gtgtgtctga atacgagtga gggaaaaggc agttccccta
      421 gacaatgtct ccctttctca gaacactgcc aagattcata ttctgggacc atgataaact
      481 gtgggaaaac aatgaataat tggcagtctc ttacagttgc ttcagaactc agagctacca
      541 tgcagttaag tgacctctgg ttctccaagg tcacaaggag gtaaataata ggagaaaggc
      601 agtaaatgaa gcattattgt gcagactctg tctgtactgt atcagttctg tagaaaaata
      661 aaggtttcct gtgttctctt cctccatggt ctcaattgtg tgattaagca cagcactgtg
      721 tggtgtgagc tgctttgccc caggctaagc actgggaggt tcagggaaac aaaaagaatc
      781 ccagaatgat gtcgtttaca taatgaaagc ttgagtaaac tgctttgtat ctatggtctg
      841 ggacatacgg agtgaccgag gggcagtccc atgcccttgc cttacagcaa gaggtgagac
      901 aaggggagaa gtggtcgtgt agcctacttt tccttttcca agatgtgtgt gcctcctcta
      961 ctcttctccc cgaatttact caggtcgtaa gagtataatt tgcaacactg tccctccaat
     1021 gcatgagcac atttgcttca aatcagtgac tgcatttcaa tatgaaatac accactggga
     1081 gaaaggagaa acaagtgttg gctagcccgg gctcgagatc tgcgatctaa gtaagcttgc
     1141 tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat
     1201 ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc attggcgaat
     1261 tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg
     1321 acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaacag cgtcaacagc
     1381 gtgccgcaga tcccaagctt gcggccgctt aagctgcaga agttggtcgt gaggcactgg
     1441 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     1501 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     1561 tgcctttctc tccacaggtg tccactccca ggttcaatac agctcttaag cggccgcaag
     1621 ctagcttgcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1681 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1741 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1801 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct tcgcccgcta
     1861 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1921 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1981 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2041 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2101 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2161 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2221 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
     2281 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2341 cgagctgtac aagtgagcct ctagactccc tcagcggcgc gcctggtgtt cctcgagatc
     2401 ggagtcctaa atcgactgcc ttagcggtag gacagtagat cagtcagcga cgtgacgaac
     2461 cctcgcagga tcaagatccc acgtgtctag agtcggggcg gccggccgct tcgagcagac
     2521 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2581 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2641 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2701 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     2761 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2821 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4801 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4861 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4921 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4981 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5041 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5101 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5161 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5221 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     5281 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5341 agcccaagct accatgataa gtaagtaata ttaaggtacg ggagg
//
