LOCUS       sequence-249927-        5323 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 5323)
  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 5323)
  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_11_Citrine
FEATURES             Location/Qualifiers
     source          1..5323
                     /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        1144..1289
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1367..1499
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1482..1501
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1553..1562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1559..1561,1562..1564,1565..2275)
                     /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(1604..1625)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1865..1884)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2212..2233
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2447..2568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2458..2477)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2512..2531
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2816..2833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2987..3575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3067..3086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3746..4537,4538..4606))
                     /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     4369..4388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4607..4711)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4738..5193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4825..4844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5035..5056
                     /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 caggtcctgg tctttaagct ctggacttcc tttttacatt taccactgta tagttcataa
      241 ttgtaatgag aatatttttc ccctcttgat gtaacatctc agggatcagc aaatctaggt
      301 ggtctttcca cctttaaaag aactgaaaaa ttttaatgga aattggagaa tagatgaatg
      361 agctaacagc agggttcaat tttaatgaca gcaacaagga attctgtaat taagcacatt
      421 aaggcagaac accctctttg aatttcagaa catctctagt tcatgctcta acatctgttt
      481 ttcttagact gctttaatct gcatcctccc ttcaggcatc ttacattaag ggtttgacag
      541 actttcctag ctgacagtag ctggcatcct tttaattgac aagggagacc actgttgaaa
      601 gagattggga agagcattct cttttccctc aaggtctaaa tgctctgctt ttgcagaact
      661 cctgcaggac ctgctagcta cagactcaac ctgaaaaggc agtttttgca gaatccaatg
      721 aagagctcaa ataagcttca catttgttcc actctctttt attttggatt tttgtcagat
      781 atcccaagag aaaaagtttt gtgttttatt ctccacatat gtgttatttt agcagaaagt
      841 gggataatct ccattaaaaa taataaaaat actgatgcat ctgcttacat ctcattaaaa
      901 tcagatacaa attaggctat ctaaaattct tatatatttg gttatgagcc tgaaacaacc
      961 ttttttttgt ttgtttgttt ttgtttttgt ttttgttttc ctgcattgtg gtcactctgt
     1021 tggctagccc gggctcgaga tctgcgatct aagtaagctt gctgcttcat ccccgtggcc
     1081 cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt ctttagttct
     1141 atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag
     1201 tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac
     1261 accgagcgac cctgcagcga cccgcttaac agcgtcaaca gcgtgccgca gatcccaagc
     1321 ttgcggccgc ttaagctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt
     1381 tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt
     1441 gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg
     1501 tgtccactcc caggttcaat acagctctta agcggccgca agctagcttg ccgccaccat
     1561 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1621 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1681 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1741 cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca
     1801 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1861 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1921 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1981 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2041 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2101 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2161 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2221 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgagc
     2281 ctctagactc cctcagcggc gcgcctggtg ttcctcgaga tcggagtcct aaatcgactg
     2341 ccttagcggt aggacagtag atcagtcagc gacgtgacga accctcgcag gatcaagatc
     2401 tcacgtgtct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     2461 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2521 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2581 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2641 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     2701 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2761 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     2821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3181 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3421 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4741 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4801 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4861 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4921 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4981 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5041 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5101 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5161 cgaattttaa caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact
     5221 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     5281 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgc
//
