LOCUS       sequence-259317-        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"
     misc_feature    14..105
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     54..73
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1366..1511
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1589..1721
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1704..1723
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1775..1784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1781..1783,1784..1786,1787..2497)
                     /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(1826..1847)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2087..2106)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2434..2455
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2671..2792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2682..2701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2736..2755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3040..3057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3211..3799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3291..3310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3970..4761,4762..4830))
                     /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     4593..4612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4831..4935)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4962..5417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5049..5068)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5259..5280
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5548..5596
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggcgcat ctttgggtgc agaacccgag tgtagatcgg acttcggatc
      181 gtgctcctat tagcaaacca aacaccggtt ttgtttttgt tgctgacgtt gtttgttttc
      241 attgccgtct gaatgagcct tttatggccc ctgccttgcc tggctgactg cacccgattg
      301 tatttaatta catttactcg gcagtttcag ccgcgatccc cggccgttac ccatcaagtc
      361 caatcactat tctttaaatc caatggctca gaatcagaga atttatcttt tgcacagtta
      421 ttgcccaaac atccagcgta aacaggcggg gctccccggc cttgctccgg ctgcgctgat
      481 taaaggccgg cagtaagatg gatggaggcg gtgcgggcag caggctcagc gccccgctgc
      541 gccccagtct ctgccattgt gcctgcctct gggtaccaga ctcgccgtgg tcctcccagc
      601 tccaaaagga aagctggctt tgtttgcccg taagcatctg caaactgttc tgacacatga
      661 tctaccctct cagcaaggcc actcattagg ctgattatca cttcaaagtt tgtattatcg
      721 tccatctcta aaggcaatta ggaatgggtg gctttgagta tcagacactg ggtgacacta
      781 agtgactcat ctttattgat tcttttgtgt gcggagacag tgattgagat gataggattg
      841 cagcctacct gcccctttga ctggtaaata ctgaagaaaa acggtgtgta attaactgac
      901 agttaatata ggttctgttt agtgttcaga aagcacaata gtggaagctg cccaaaagga
      961 acaaagtaat gaaactcctt tgacaagtta aactctaaag ccaactgtgt aattatctga
     1021 cggttaacgg ggggagggga ggactcggat tcgtgtttat gagaccaaag gaattcctgc
     1081 ccctgaacaa tggggacagt tactgacaat aatcataaca gctacaccag atatttgttt
     1141 gtgcaagcct ggcttgcagg cgagctcagc ccggtaccca accccgctcc ccgtctgtgg
     1201 gtcagggata ccgagcccag gaaaccacag cctgttctct gttggctagc ccgggctcga
     1261 gatctgcgat ctaagtaagc ttgctgcttc atccccgtgg cccgttgctc gcgtttgctg
     1321 gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg
     1381 cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc
     1441 aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc
     1501 gacccgctta acagcgtcaa cagcgtgccg cagatcccaa gcttgcggcc gcttaagctg
     1561 cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
     1621 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1681 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact cccaggttca
     1741 atacagctct taagcggccg caagctagct tgccgccacc atggtgagca agggcgagga
     1801 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1861 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1921 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     1981 cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2041 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2101 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2161 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2221 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2281 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2341 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     2401 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2461 cgccgggatc actctcggca tggacgagct gtacaagtga gcctctagac tccctcagcg
     2521 gcgcgccgat ctgcggttgc cgaggtatct tccgggtatt atgcgccatt gcgccacgga
     2581 ctagcatcct gctagattcg cgtcttgata cgggcctaaa gatctcacgt gtctagagtc
     2641 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2701 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2761 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2821 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2881 gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2941 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     3001 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     3061 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3121 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3181 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3241 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3301 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3361 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3421 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3481 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3541 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3601 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3661 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3721 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3781 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3841 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3901 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3961 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4021 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4081 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4141 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4201 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4261 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4321 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4381 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4441 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4501 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4561 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4621 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4681 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4741 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4801 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4861 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4921 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     4981 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5041 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5101 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5161 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5221 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5281 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5341 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5401 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5461 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     5521 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     5581 gttggttttt tgtgtg
//
