LOCUS       sequence-258406-        6653 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 6653)
  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 6653)
  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_Zic1_enh_373a_Citrine
FEATURES             Location/Qualifiers
     source          1..6653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      30..39
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(36..38,39..41,42..752)
                     /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(81..102)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(342..361)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     689..710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    926..1047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(937..956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     991..1010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1295..1312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1466..2054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1546..1565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2225..3016,3017..3085))
                     /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     2848..2867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3086..3190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3217..3672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3304..3323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3514..3535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3803..3851
                     /note="synthetic polyadenylation signal"
     misc_feature    3865..3956
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3905..3924
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        6274..6419
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          6497..6629
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6612..6631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gctcttaagc ggccgcaagc tagcttgccg ccaccatggt gagcaagggc gaggagctgt
       61 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      121 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      181 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc
      241 tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      301 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      361 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      421 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      481 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
      541 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
      601 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
      661 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
      721 ggatcactct cggcatggac gagctgtaca agtgagcctc tagactccct cagcggcgcg
      781 ccgttgcgtt ccaaacgtcg tggagctccc tgtcaagaca cgaaccgcta acgccacctt
      841 agtccgtgaa gcatgatacg cgacataccc caggagatct cacgtgtcta gagtcggggc
      901 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
      961 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1021 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1081 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     1141 atcgataagg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     1201 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     1261 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1321 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1381 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1441 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1501 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1561 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1621 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1681 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1741 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1801 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1861 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1921 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1981 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2041 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2101 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2161 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2221 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2281 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2341 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2401 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2461 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2521 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2581 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2641 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2701 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2761 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2821 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2881 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2941 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3001 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3061 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3121 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3181 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     3241 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3301 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3361 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3421 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3481 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3541 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3601 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3661 cgcttacaat ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3721 gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac
     3781 gggaggtact tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     3841 ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa
     3901 caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg
     3961 ataggtaccg agctcgagtt gccaggtgct cctatgctgt atacccaagg agaacagtct
     4021 gtcattccac aaactctcct gtattttatg gactcaacat tgctattcat ctcaggcaac
     4081 tgcagccaat cttggctgcc aggtgaaaca tgatgttgaa acagagaaac agtcttgaag
     4141 aaatacaagc tcttaaacca tgaatggctt tgtggatcaa aaatgccttt ttgaattaca
     4201 cctgaaaatg gagatcagca aaatctctgg agcatggtga aggaggagtc gtgccggaca
     4261 ggcagtccca cccttgcctg ccaagcattt tgcaggtctg ctctaagcac atcagttatg
     4321 attaggagca gctgcaaact cagcccagtg cagtgctccc aataaattcc agatgatgaa
     4381 tatgatgcag acttactgga agaaaaatat atttctgaag agaaaaacaa ttacttctga
     4441 aagcaagaca atggaactga gctgatgtga tgctgggcat aagatagaac agaactttag
     4501 ggcaatggct ttaacttcta aaaaatcacc cacctggaaa gccttttcta gactgtattt
     4561 ggattcctca gacctaaagt actttcagag ttaaacttga aaaattatgc atttcactct
     4621 tgatttctcc cgaaaaggac agttgctttg ttctgtcatt atctcacagt tttatgatta
     4681 gcagaaaggt gcctttgcta cccccgaaca acatgtatgt cgtttcacag aagtgcaaaa
     4741 aggcttagta acaccacttg atgcagtggg tttcattggc tgaatcttcg gcggggactt
     4801 gaaggaaatt tgcaaaaaac atgtcaaaag ttaagtggtg aaaatcagct taaatgccta
     4861 attttaattt tgtttgtgct ctttgttgtt tcattgtagg gagagggaag aagctgtctt
     4921 tctctctgtc ctttcatttt ggttgggaga aaatggatgt tggtgaaaag aagagagctc
     4981 tgacagggac tgggaagggg ttgtttattc tgttctcttt tctttagaaa ggggcaactc
     5041 ttctaaaact ggtgatgatc cagttagagg gggacttggg aggggagaga aaagaaaact
     5101 aataatcagc ctctaaatga caagtttcct cttgaacaaa agagcaacag gaggcagtgc
     5161 aggcgggggt ccatgcccag gggcaacgtc gccaaaacaa tggctggaga atgtgcagct
     5221 tttgccactc caggatctac tgccccagtg tgtttgctgg caggctggaa ggatccaacc
     5281 cgaccaaagg ctgggtgatg tgctgatgcc acttctggag tcctccgggg tgaaatgatt
     5341 gagtgctagt aaggtaaaca aggaaagagg actgcaagag agtctggccc agaatgacaa
     5401 gtatgaatag cctcaacata gcccctgtca gtgaagggac tacaggtggg cctcagagtg
     5461 agtttaatta aagatgacta ctgctcttta atatctgtgt ttctcttgca tttattaact
     5521 gtcttctggg agacactacc agtgccagat aaactgaaat gagatgattt taaattattg
     5581 agtgttcaca cagaagttca ttctggttta gactaaattg gttttaattt aaattcatgt
     5641 aacttctgtg tagatgaggc ctatggtttg ccattctgca gcagtggtga atttatagca
     5701 tgcttgctcc aaacgtgctg tcatgcaaat tctcttggca tttagtccaa gtccagttcc
     5761 agctgttggt ggagatggag gagcagaatg gactgctcac atggcctccc atgtaatgca
     5821 gattggatct attgtcagtc agatggactg gtgttgtggg agcagtgtga agccagttct
     5881 ttcactacat ggtgtgtaaa gcaatgaggc agacaaaagc tggatttaaa ttagggaata
     5941 tttctgccaa atttcattct ctgaagcaca gaaataaaac gtttaatgca tctggcaatg
     6001 tttattctgt gcacagtgtt ctagctgatc ataaaagaaa agctacgtgt gaacattatc
     6061 tagaaacttc atcgtattca gagaagagag agcatgattt tgctgcagtc tctgaaaaag
     6121 tatttgactg tgaaagcgga tacagtctgt tggctagccc gggctcgaga tctgcgatct
     6181 aagtaagctt gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg
     6241 gaagaaatat atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg
     6301 tcattggcga attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg
     6361 catattaagg tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaac
     6421 agcgtcaaca gcgtgccgca gatcccaagc ttgcggccgc ttaagctgca gaagttggtc
     6481 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     6541 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     6601 tgacatccac tttgcctttc tctccacagg tgtccactcc caggttcaat aca
//
