LOCUS       sequence-249941-        5596 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 5596)
  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 5596)
  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_Msx1_enh_264_Citrine
FEATURES             Location/Qualifiers
     source          1..5596
                     /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        1415..1560
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1638..1770
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1753..1772
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1824..1833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1830..1832,1833..1835,1836..2546)
                     /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(1875..1896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2136..2155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2483..2504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2720..2841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2731..2750)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2785..2804
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3089..3106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3260..3848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3340..3359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4019..4810,4811..4879))
                     /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     4642..4661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4880..4984)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5011..5466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5098..5117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5308..5329
                     /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 caggcgcatc tttgggtgca gaacccgagt gtagatcgga cttcggatcg tgctcctatt
      241 agcaaaccaa acaccggttt tgtttttgtt gctgacgttg tttgttttca ttgccgtctg
      301 aatgagcctt ttatggcccc tgccttgcct ggctgactgc acccgattgt atttaattac
      361 atttactcgg cagtttcagc cgcgatcccc ggccgttacc catcaagtcc aatcactatt
      421 ctttaaatcc aatggctcag aatcagagaa tttatctttt gcacagttat tgcccaaaca
      481 tccagcgtaa acaggcgggg ctccccggcc ttgctccggc tgcgctgatt aaaggccggc
      541 agtaagatgg atggaggcgg tgcgggcagc aggctcagcg ccccgctgcg ccccagtctc
      601 tgccattgtg cctgcctctg ggtaccagac tcgccgtggt cctcccagct ccaaaaggaa
      661 agctggcttt gtttgcccgt aagcatctgc aaactgttct gacacatgat ctaccctctc
      721 agcaaggcca ctcattaggc tgattatcac ttcaaagttt gtattatcgt ccatctctaa
      781 aggcaattag gaatgggtgg ctttgagtat cagacactgg gtgacactaa gtgactcatc
      841 tttattgatt cttttgtgtg cggagacagt gattgagatg ataggattgc agcctacctg
      901 cccctttgac tggtaaatac tgaagaaaaa cggtgtgtaa ttaactgaca gttaatatag
      961 gttctgttta gtgttcagaa agcacaatag tggaagctgc ccaaaaggaa caaagtaatg
     1021 aaactccttt gacaagttaa actctaaagc caactgtgta attatctgac ggttaacggg
     1081 gggaggggag gactcggatt cgtgtttatg agaccaaagg aattcctgcc cctgaacaat
     1141 ggggacagtt actgacaata atcataacag ctacaccaga tatttgtttg tgcaagcctg
     1201 gcttgcaggc gagctcagcc cggtacccaa ccccgctccc cgtctgtggg tcagggatac
     1261 cgagcccagg aaaccacagc ctgttctctg ttggctagcc cgggctcgag atctgcgatc
     1321 taagtaagct tgctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
     1381 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
     1441 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
     1501 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     1561 cagcgtcaac agcgtgccgc agatcccaag cttgcggccg cttaagctgc agaagttggt
     1621 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     1681 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     1741 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccaggttcaa tacagctctt
     1801 aagcggccgc aagctagctt gccgccacca tggtgagcaa gggcgaggag ctgttcaccg
     1861 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1921 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1981 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
     2041 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2101 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2161 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2221 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2281 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2341 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2401 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     2461 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2521 ctctcggcat ggacgagctg tacaagtgag cctctagact ccctcagcgg cgcgccgatc
     2581 tgcggttgcc gaggtatctt ccgggtatta tgcgccattg cgccacggac tagcatcctg
     2641 ctagattcgc gtcttgatac gggcctaaag atctcacgtg tctagagtcg gggcggccgg
     2701 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2761 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2821 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2881 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     2941 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     3001 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     3061 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3421 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     3481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3661 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4981 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     5041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5401 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5461 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5521 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     5581 tacttggagc ggccgc
//
