LOCUS       sequence-258987-        5317 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 5317)
  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 5317)
  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..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1076..1221
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1299..1431
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1414..1433
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1485..1494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1491..1493,1494..1496,1497..2207)
                     /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(1536..1557)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1797..1816)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2144..2165
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2379..2500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2390..2409)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2444..2463
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2748..2765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2919..3507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2999..3018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3678..4469,4470..4538))
                     /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     4301..4320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4539..4643)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4670..5125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4757..4776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4967..4988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5256..5304
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cgagttgcca
      121 ggtcctggtc tttaagctct ggacttcctt tttacattta ccactgtata gttcataatt
      181 gtaatgagaa tatttttccc ctcttgatgt aacatctcag ggatcagcaa atctaggtgg
      241 tctttccacc tttaaaagaa ctgaaaaatt ttaatggaaa ttggagaata gatgaatgag
      301 ctaacagcag ggttcaattt taatgacagc aacaaggaat tctgtaatta agcacattaa
      361 ggcagaacac cctctttgaa tttcagaaca tctctagttc atgctctaac atctgttttt
      421 cttagactgc tttaatctgc atcctccctt caggcatctt acattaaggg tttgacagac
      481 tttcctagct gacagtagct ggcatccttt ttattgacaa gggagaccac tgttgaaaga
      541 gattgggaag agcattctct tttccctcaa ggtctaaatg ctctgctttt gcagaactcc
      601 tgcaggacct gctagctaca gactcaacct gaaaaggcag tttttgcaga atccaatgaa
      661 gagctcaaat aagcttcaca tttgttccac tctcttttat tttggatttt tgtcagatat
      721 cccaagagaa aaagttttgt gttttattct ccacatatgt gttattttag cagaaagtgg
      781 gataatctcc attaaaaata ataaaaatac tgatgcatct gcttacatct cattaaaatc
      841 agatacaaat taggctatct aaaattctta tatatttggt tatgagcctg aaacaacctt
      901 ttttgttttg ttttgttttt gtttttgttt tcctgcattg tggtcactct gttggctagc
      961 ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc atccccgtgg cccgttgctc
     1021 gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac
     1081 acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg
     1141 gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg
     1201 accctgcagc gacccgctta acagcgtcaa cagcgtgccg cagatcccaa gcttgcggcc
     1261 gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1321 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1381 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1441 cccaggttca atacagctct taagcggccg caagctagct tgccgccacc atggtgagca
     1501 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1561 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1621 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1681 ccttcggcta cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1741 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1801 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1861 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1921 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1981 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2041 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     2101 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2161 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga gcctctagac
     2221 tccctcagcg gcgcgcctgg tgttcctcga gatcggagtc ctaaatcgac tgccttagcg
     2281 gtaggacagt agatcagtca gcgacgtgac gaaccctcgc aggatcaaga tcccacgtgt
     2341 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2401 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2461 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2521 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2581 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2641 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2701 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2761 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2821 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2881 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2941 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3001 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3061 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3121 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3181 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3241 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3301 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3361 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3421 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3481 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3541 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3601 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3661 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3721 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3781 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3841 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3901 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3961 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4021 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4081 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4141 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4201 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4261 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4321 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4381 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4441 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4501 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4561 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4621 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5041 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5101 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     5161 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     5221 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     5281 atctgtgtgt tggttttttg tgtgaatcga tagtact
//
