LOCUS       sequence-249956-        5842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enhancer reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5842)
  AUTHORS   Williams RM, Candido-Ferreira I, Repapi E, Gavriouchkina D, 
            Senanayake U, Ling ITC, Telenius J, Taylor S, Hughes J, 
            Sauka-Spengler T
  TITLE     Reconstruction of the Global Neural Crest Gene Regulatory Network In
            Vivo.
  JOURNAL   Dev Cell. 2019 Oct 21;51(2):255-276.e7. doi: 
            10.1016/j.devcel.2019.10.003.
  PUBMED    31639368
REFERENCE   2  (bases 1 to 5842)
  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: 
            "10.1016/j.devcel.2019.10"; journalName: "Dev Cell"; date: 
            "2019-10-21- 21"; volume: "51"; issue: "2"; pages: "255-276.e7"
COMMENT     Sequence Label: pTK_Axud1_enh_394_Citrine
FEATURES             Location/Qualifiers
     source          1..5842
                     /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        1661..1806
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1884..2016
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1999..2018
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2070..2079
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2076..2078,2079..2081,2082..2792)
                     /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(2121..2142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2382..2401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2729..2750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2966..3087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2977..2996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3031..3050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3335..3352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3506..4094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3586..3605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4265..5056,5057..5125))
                     /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     4888..4907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5126..5230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5257..5712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5344..5363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5554..5575
                     /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 ctcgagttgc
      181 caggcaaaca gagcccgaga gaggaaaatg aagctctgcc tattttacag gcagattttt
      241 gcaacagtgc agatgtcagc cttgctgttt cagaacagcc atgccctgcc ctataaacgt
      301 gtctcagata catcacattg ttgtgggtct gggaaactgg gagttgctgg tccttgccta
      361 ccctctctac caccattaaa gcaaatccca tcctcagaaa agagggaatt cttagaaacc
      421 tttgccttgc accagcaaca tcctgcaaac tcgaacgggt taatcccctt tagccagctt
      481 acccataatg aaaggtttca ctctttgtca cctaatggca gccaggtctt gtgtctaaaa
      541 tacaagccta gctgcctttg catgaaatgt ttttaatcat cgtggaataa aaaccttgcc
      601 tggctagtgc caaccaaaga gatgtttgtc tgctcagcag attttatcaa agcttccttg
      661 atgtctctgc ctttgtaatg ctaagcgctt cactgtttgg catatttaaa gtaggatgcg
      721 gtacatgtga agcatttcac agagcgatga tgtatcgagt gtgctttagg aagtaatgaa
      781 agcagcagac tcatgatttt tgagctttta attacttatt aagtggcatg acataaaggc
      841 tacgggggag attttttttc agcatccaac ttactgactt tgaagtcagt gcctaaatac
      901 cctttgtgcc tttgaaactt tccgtctgct cacaggggaa cgcgctgccg ctctgggctt
      961 cgtttcacgc tgaccacatt tgttcagggc aggggatggg agtctgctca gacacaggtc
     1021 tctgccttcg aggtggtgga taaaacgagg aacagactaa agacgtgcct tcggacaaag
     1081 gatggggaga gcctggagag agcgcagagc ctggagcaat gagttctgac ctggcctgaa
     1141 aaatgccagc actacagggg gcaaataacc atagcagccc ccggggtgct tttaatgagc
     1201 tgaattgcag tcttgagtcc agccttccat cactcacaag ccgctgccac agctggtttt
     1261 agcagatgtc tcctccgaca gaaaggatgc ggcaaagcac agtggttgac ctcgcctcca
     1321 tctcctctgg gtccgctccg accgctgtga ctacaaataa aagataaagt caggaaaggc
     1381 caaaaagaag aaaagattct cagatttgtg gttgtgttta ttggagtttc ttttcttttt
     1441 gtttgtttgt ttctttgttt tgcttttttt ttttctgtgt ttttttccct tgtttttgac
     1501 tggtttttca tatgcaggtt ttatggggag tgactgttgg ctagcccggg ctcgagatct
     1561 gcgatctaag taagcttgct gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt
     1621 gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa ccccgcccag
     1681 cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc
     1741 cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc
     1801 gcttaacagc gtcaacagcg tgccgcagat cccaagcttg cggccgctta agctgcagaa
     1861 gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc
     1921 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     1981 gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag gttcaataca
     2041 gctcttaagc ggccgcaagc tagcttgccg ccaccatggt gagcaagggc gaggagctgt
     2101 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2161 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2221 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc
     2281 tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2341 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2401 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2461 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2521 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2581 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2641 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
     2701 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2761 ggatcactct cggcatggac gagctgtaca agtgagcctc tagactccct cagcggcgcg
     2821 ccgttgcgtt ccaaacgtcg tggagctccc tgtcaagaca cgaaccgcta acgccacctt
     2881 agtccgtgaa gcatgatacg cgacataccc caggagatct cacgtgtcta gagtcggggc
     2941 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3001 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3061 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3121 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     3181 atcgataagg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     3241 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     3301 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     3721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     5281 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5341 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5401 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5461 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5521 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5581 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5641 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5701 cgcttacaat ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5761 gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac
     5821 gggaggtact tggagcggcc gc
//
