LOCUS       sequence-249907-        6897 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 6897)
  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 6897)
  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_Snai2_enh_242_Citrine
FEATURES             Location/Qualifiers
     source          1..6897
                     /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        2716..2861
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2939..3071
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3054..3073
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3125..3134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3131..3133,3134..3136,3137..3847)
                     /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(3176..3197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3437..3456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3784..3805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4021..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4032..4051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4086..4105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4390..4407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4561..5149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4641..4660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5320..6111,6112..6180))
                     /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     5943..5962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6181..6285)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6312..6767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6399..6418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6609..6630
                     /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 caggcactta ggtagatgca ggtccagata tccatagcgc attaagagat gcctttagcc
      241 agctgaaaac tctttttgat ctctctgata ataccaggca agcactgatc tgaaaaggca
      301 gattaccacc aaggattaag cagtacaata gcatgctgag aatgtaagag aatgatgagg
      361 tttagcaaac aagcgtggaa catatttgaa ctcaacatcg aacaccaaaa gaagacatag
      421 gaagagaact gaggctgtag ggaactgccc tgaaaaatgt tatatgagag agggcaaaac
      481 tgtctaacta ctagatttta acttaaaatc aggactcagc tagtcaagaa aacacattct
      541 gcacaaaaaa tctcttctca gtgctgaagg cacagtctga atggacacag agtctcagct
      601 acactttaat tgtcctgaga ctatcctttc aggcatgtgg ctggagcagg tcacactgct
      661 gtagctgctg tcctcaggca gtaagacaca gtgaagacag ggctttacaa agaccttttt
      721 ttttaaagag cttttttttt tttttttctt tttcaggact agtcacagta aatagagtgg
      781 ttatttggta tgtgagataa gaattcgcca acatcatttg aacacactag ttttctttca
      841 gtgttttgtc acttgaccag tttaatgcta aggttaaaaa gctcttacac aacatgtatc
      901 caccctcttg tgtgtcaaaa atgtgaggaa tgtttgcaag aaacagcagc ttgcgtacaa
      961 ttcctgtcac caaagcccaa agttgaaatg gctaacgttt cctggatctg cttagctcac
     1021 ctacggagac agagcttgcc gtgcctgtgt gcaaacagaa gcagagtcat taaaacggga
     1081 gcagttcttg ggcaagctcc agtttgcatg ttacatattc ataaatagct gtgttagaat
     1141 ggcttccaac agtgcaaacg tcaatggagc ctgctggagc aagccttagg tatgccatct
     1201 atgtgtctga ttgcctggag ttcagaggaa cttgctgaat gtattaccaa caagaatgaa
     1261 aaggcttgca tcagggtagc tggccaggta caattgctgg gtatttaaaa ggacgtatca
     1321 gtaccaaatt gtttcacagc tgtaattttt aaactcactt gctgtttgtg taatgcttga
     1381 gaatctccaa caggaatatt tcattctgtt gttctttcaa agagcacata ttaatttttg
     1441 ttgtaacaaa aaattttcat tagggcttgg ctaatagcac tgattatttg tttattttca
     1501 ctctgtgcaa cctatttaat tgtgtataca catggctata attaccactt tgaaatgcaa
     1561 atacacagtt actaaggata acccttcttg ttttcaagta gctaacactc ctttttcata
     1621 aatgctaacc aaatcataag atcccagtaa aacagcacag tttaagctct ccccacattg
     1681 caatggctgc ctcctgctcc aaattactct ctctgccact gtcatagctt ccttcaattt
     1741 atcacgtcag aggccctagg caagggccac gaggtcattt gaccaaagta ctggccagct
     1801 tacagctgag tgtacaccag gactcctcta agtgatagga agaagagtga gtgcgtttta
     1861 tgccaaactg tgtctcatcc agtctccaac tccatttgta acctatccac ggaaaggtct
     1921 gtgttggtac agagtcattt gttactctca gttattcacc agatagttta ttgctattta
     1981 tttccaccca cagactgctc tcagactttc ctctgggaca ggatttgctt cctaaaggga
     2041 ggatttaggg atttgactaa gcagcatgag cagcagaaga tatcacataa caatggcttt
     2101 acttcctaaa tgtcaaactc acaaagagct ttcaaagtga ccgtaccaac ccaccttcct
     2161 tttcgcatca cttccccttg cccccaatca attaccgatt tcagtggtta cccatagaag
     2221 tttctctttc agcagcattt ttgtgagcaa atatttgctt gtccatctat tcacagagag
     2281 cacattaaag cagcgtgcca gttagagtgt gggctgtctt tgaacagatg ttatgaatac
     2341 tgttttgcag gatttgccca gcagtagatg agaacaagta atgaggaaac taggaacatg
     2401 ggtaacgagg ccagaaggtg tggaagcttt ccaggagcag tgggagctgg gtggagaaaa
     2461 cttgacttac tgatatgcag ccacgagccc ttctcagact ttatttgttc ttcttaatgg
     2521 tcactctcaa gaaaagttat tcatcagggc tcattccctt gagaattaat tttcctgtgc
     2581 cttcgaggct gttggctagc ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc
     2641 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     2701 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     2761 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     2821 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     2881 cagatcccaa gcttgcggcc gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     2941 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     3001 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     3061 tctctccaca ggtgtccact cccaggttca atacagctct taagcggccg caagctagct
     3121 tgccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3181 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3241 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3301 ctggcccacc ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc gctaccccga
     3361 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3421 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3481 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3541 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3601 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3661 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3721 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3781 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3841 gtacaagtga gcctctagac tccctcagcg gcgcgccgtt gcgttccaaa cgtcgtggag
     3901 ctccctgtca agacacgaac cgctaacgcc accttagtcc gtgaagcatg atacgcgaca
     3961 taccccagga gatctcacgt gtctagagtc ggggcggccg gccgcttcga gcagacatga
     4021 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4081 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4141 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4201 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg
     4261 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     4321 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     4381 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4501 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4561 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4621 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4681 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4741 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4801 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4861 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4921 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4981 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5041 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5101 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5161 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5221 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5281 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5341 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5401 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5461 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5521 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5581 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5641 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5701 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5761 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5821 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5881 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5941 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6001 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6061 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6121 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6181 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6241 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6301 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6361 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6421 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6481 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6541 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6601 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6661 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6721 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt
     6781 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc
     6841 caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgc
//
