LOCUS       sequence-258984-        5912 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 5912)
  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 5912)
  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_Zfhx4_enh_5_Citrine
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1336..1481
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1559..1691
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1674..1693
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1745..1754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1751..1753,1754..1756,1757..2467)
                     /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(1796..1817)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2057..2076)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2404..2425
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2639..2760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2650..2669)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2704..2723
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3008..3025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3179..3767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3259..3278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3938..4729,4730..4798))
                     /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     4561..4580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4799..4903)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4930..5385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5017..5036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5227..5248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5516..5564
                     /note="synthetic polyadenylation signal"
     misc_feature    5578..5669
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5618..5637
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aaggtataga tctgaaagaa tagaatttta tgttatcatt ttaaatatat atacaaatga
       61 ctcaccctgt tttcatcatg aaattccctg aagagtgaag ctgtaaaacc tcctgagaaa
      121 tttcctgaag acataaaggc cttagaatgt tgcttgaagc cactgccaat tcattactca
      181 acagaagcaa tcagtgtttc atctccagaa gaaaagacaa aaaaaaaaaa aaaaaaaatt
      241 ctcttacctc ctagggactt cctgaaagta ggaagagaaa gagaaaactg agagaacact
      301 actggctcag tgcttccaca aagcactcct ttttccttct ctcagctgaa tgcaaatgac
      361 ctcttctagt gtggaaggtc acctcatcgc ccagagaaaa gcagaagacc agaattcact
      421 gtacactcat tgcactcacc ctcattgtac agcaacattg gttgtttgga catctaggaa
      481 gggaaagagg acagatctgt atcactctat cgccttgcaa ttcttcgttt cccttattca
      541 cagtaaaatg gagaaatgca tgcttagaag agatcgcttt ttcccttatt cctagtaaca
      601 gagatcaatg gcaaataata cagagactag cacatttata ggattcaacc tcagaaatag
      661 ttttctgctc tactgtactt tggtttggct gattttatgt acaaaagaga atattcaata
      721 ttcaaatgga tgccaagcac agtagtatag tgctttaacc tttccctttg aaatatttta
      781 gttcagtggg tggcagagaa aaccacaaac tgtggcatga ctgtcagtct cataaatgat
      841 ttgtgactgt gtaaatgcag atgccacagc tgtcatgtaa tctgcacaca ggctatactg
      901 agaatgtaac tcagaccctt ctgctccagg aacacaggcc ttttccattt gtactgttcc
      961 attagctgtt gaaatggctt aaaactagaa caatttatcc ctttcccttt tgaactttag
     1021 taaaatagta cagagctctg aaatggcttt catttataaa ccaaaattct cccaatgtga
     1081 ttttctaaag caaacattta aatccagtga tatctaaata gctgtcaaca gtattatagg
     1141 gagctgtaga aaactttaaa agtaaaactc agaaatggat acattttcct ggtttcaagt
     1201 gtcattgtct gttggctagc ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc
     1261 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     1321 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     1381 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     1441 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     1501 cagatcccaa gcttgcggcc gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     1561 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     1621 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1681 tctctccaca ggtgtccact cccaggttca atacagctct taagcggccg caagctagct
     1741 tgccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1801 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1861 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1921 ctggcccacc ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc gctaccccga
     1981 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2041 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2101 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2161 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2221 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2281 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2341 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2401 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2461 gtacaagtga gcctctagac tccctcagcg gcgcgcctgg tgttcctcga gatcggagtc
     2521 ctaaatcgac tgccttagcg gtaggacagt agatcagtca gcgacgtgac gaaccctcgc
     2581 aggatcaaga tcccacgtgt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     2641 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2701 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2761 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2821 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     2881 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     2941 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     3001 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3061 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3121 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3181 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3241 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3301 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3361 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3421 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3481 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3541 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3601 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3661 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3721 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3781 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3841 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3901 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3961 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4021 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4081 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4141 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4201 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4261 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4321 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4381 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4441 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4501 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4561 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4621 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4681 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4741 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4801 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4861 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4921 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4981 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5041 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5101 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5161 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5221 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5281 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5341 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca
     5401 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca
     5461 agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa
     5521 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac
     5581 atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag
     5641 tgcaagtgca ggtgccagaa catttctcta tcgataggta ccgagctcga gttgccaggt
     5701 gcgaggttca aggaaaacta tttgttcaac tgaaaacagg caggaagaag gatgtgggaa
     5761 tttagagaca gactttacat tcaagactcc tcagaatggg aaaaacatag ataagaaaaa
     5821 cttctctttg aactattaga gatactctta aaaatatagg ctgtgttttt ccataacgtt
     5881 atggttatgg aaaataatat tacttttaat tg
//
