LOCUS       sequence-258446-        6710 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 6710)
  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 6710)
  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_FoxD3_enh_193_Citrine
FEATURES             Location/Qualifiers
     source          1..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2375..2520
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2598..2730
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2713..2732
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2784..2793
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2790..2792,2793..2795,2796..3506)
                     /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(2835..2856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3096..3115)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3443..3464
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3680..3801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3691..3710)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3745..3764
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4049..4066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4220..4808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4300..4319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4979..5770,5771..5839))
                     /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     5602..5621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5840..5944)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5971..6426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6058..6077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6268..6289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6557..6605
                     /note="synthetic polyadenylation signal"
     misc_feature    6619..6710
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6659..6678
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atcgataggt accgagctcg agttgccagt ggaccactct gtgttctgtg tattgcacca
       61 gagcagtaac aggtatgctt tcataagcag ggtgtaatag gagtgatgtg attcagggtt
      121 tgcccttacc atttgttgca taccatccac tctggtgttg tcacaacgac tcttagacat
      181 actgatcttg caaaacacag ggccacccac agccagattc agcataaagc aaaatcagtg
      241 cacagaacct aagagaaacg atggctgtgc aatgctcagg tgagaaatcg ttccacccaa
      301 atggaccatg aatgtaaaca ctgactaacg agagtggtac agattgggcc attaaaggcc
      361 aggcagagat gaatggaaac gtctgagcag gaacataaaa caggccaatt tctgccacta
      421 agtaggagtg ggagtataac ggtccttgtt atagagttgc tggcagtgat gctactgtta
      481 aggctgaaaa agacatcaca ttccaaatct ctcttttcac ttgtaaaaca gtaagatttt
      541 gagcttgatc cctcacttca aaggtagcca ctgccactgc ttaacaaaga gaaacatgag
      601 atggatgaga agaaaaaaat caagccggct acaaaaggtt atagagacga cccagagtgg
      661 gaagtgtgag acttgagtta ccgtgaagtt gagttctgct tttcttttgc agcgactgat
      721 ttctcccacc tctgaaagcc aacacagaaa tagccagaga ataacccagc ttttggtttt
      781 atccatcagc aaaaggtttt catgcatcaa atctgtttgt ggattctaaa aaggctgatt
      841 catgacgtgg tttatgaaca ctgtttgttc cttcaagttc ctgattgctt ccacgggatc
      901 catttggctt ctcctgactc gagctgagag tctttccaac aatagagaga acagaggctc
      961 ggaagaactg agccacaggc tttgaagtgt gaacatttgt ctttgctctg atcagaaatg
     1021 aaacctgaaa ttttcaatga aggaaaattt gtacaatgtg ggaaacagtt tcggggcaaa
     1081 attgaaacaa taattctaac gtggcctctt agaatgtgac ctttttatct cttaacaaca
     1141 tttaaatata ataaagaatt tgaatatttg aaaagtcatt ttaacatttg aaatttccac
     1201 ttttttgctc tgaaaatgtc aaaacaggat atttcagctt tctgggaact tctttccctc
     1261 ccactttcac ttgaaaatga gaacttccag cctgtattga tcgtccatgt ggctccctgg
     1321 tacagccagc cacgagcaga aagggaaagg tgacagattg cactaggtcc tgtgacacca
     1381 aacacagcta ttcctttcct attgccattg cccaatacaa cttattattg gataggctgt
     1441 ggaggaagag gggggtaggt tgggaagcat ttctttgtta accactttgt cactgtgtta
     1501 gagacgcatg caatctgcat ttgtgattga gtctttggag tggcacagga aaatacttgg
     1561 acaagggagc aggcagagct tttgggatct tcagtgcttc atcaaaggta acagccagtt
     1621 gttttgtgct gccttcagtg cttcatgcac tgagaacact ggttgcaaag aacttccact
     1681 gcatggtgcg tgtaatctgt aattgtgtat aacttgtttg cacccagctt ccggcctgaa
     1741 gcaggcacgt taagcactgg gtgatgtggc tcccagcctt tgcaaagctc cccacattta
     1801 cgtgccacta actcaggctg cagccttgtg agcgttgctc acaccggatg acaccggaac
     1861 actttgaaga gttcctgatg gcatctgtcc ctgcagctct caccacagag gttcctgccc
     1921 ccaggttgtg tgtctgctca agctgatcca ttcacagccc agtgaagttt tgctatttag
     1981 ataattatgc aactgtttta ggcttaaaat tgactgtcag ctgttagtga gagagaaata
     2041 gcagttctaa actttcaact gctttgatta aatcaaaatg tgactgtgaa tcataggtct
     2101 caacccacag tattgccctg gtgcaaatgt ggtgcaaaga gaaatcggtt cccatcctgg
     2161 aaaaacaact gttccactca gcagtggaac acctccatcc agccatgcct cggggacagc
     2221 tcttggccaa cagggtggtc tcagttcctg ttggctagcc cgggctcgag atctgcgatc
     2281 taagtaagct tgctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
     2341 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
     2401 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
     2461 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     2521 cagcgtcaac agcgtgccgc agatcccaag cttgcggccg cttaagctgc agaagttggt
     2581 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     2641 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     2701 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccaggttcaa tacagctctt
     2761 aagcggccgc aagctagctt gccgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2821 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2881 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2941 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
     3001 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3061 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3121 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3181 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3241 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3301 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3361 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     3421 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3481 ctctcggcat ggacgagctg tacaagtgag cctctagact ccctcagcgg cgcgccacga
     3541 agctggtaga gtgctggcct tgtgccaagt tgcgggagta cccggatctg cgtttacata
     3601 cccaccactc accgtcttaa agcgggacag atctcacgtg tctagagtcg gggcggccgg
     3661 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3721 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3781 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3841 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     3901 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     3961 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     4021 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4081 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4141 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4201 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4261 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4321 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4381 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4441 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4501 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4561 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4621 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4681 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4741 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4801 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5941 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     6001 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     6061 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     6121 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     6181 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6241 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6301 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6361 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     6421 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6481 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     6541 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     6601 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     6661 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
