LOCUS       sequence-258404-        4957 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 4957)
  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 4957)
  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_Lmo4_enh_15_Citrine
FEATURES             Location/Qualifiers
     source          1..4957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        608..753
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          831..963
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     946..965
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1017..1026
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1023..1025,1026..1028,1029..1739)
                     /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(1068..1089)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1329..1348)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1676..1697
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1911..2032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1922..1941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1976..1995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2280..2297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2451..3039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2531..2550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3210..4001,4002..4070))
                     /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     3833..3852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4071..4175)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4202..4657
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4289..4308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4499..4520
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4788..4836
                     /note="synthetic polyadenylation signal"
     misc_feature    4850..4941
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4890..4909
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ctcgagttgc caggtctttg agcaatggaa gccttcgctt ctaggggtgt tgtgagaggg
       61 caatgctacc cagtcggcac acggaggtgt tcggtagggc tcgttttggt catcgtcctg
      121 agcagcaagc cgaagcccgg cagaatggta attaaataga gcagaacggt gctgggaagg
      181 agcctcgttt gaagtggagg aattgacgct gctcttgggc tgcagagtgg tgcggatcca
      241 tcactcggca gcaccatcaa acaaacctgt gtctgattgt tcctaattaa agggagagga
      301 tgtgtccacc gtgtcccaga ggcaagttca ggcagttatc gcaaccccct ggtttcgtgt
      361 tgccaaaaat taatactcgt ttattttaac gtaggcggtt ataaatagga tgctttatca
      421 gacagggatt tgaattgatc tcctcgttgt atcgcccgcc ctcccttaga tccgtgccat
      481 ctgttggcta gcccgggctc gagatctgcg atctaagtaa gcttgctgct tcatccccgt
      541 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
      601 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat
      661 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
      721 gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc
      781 aagcttgcgg ccgcttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
      841 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
      901 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
      961 caggtgtcca ctcccaggtt caatacagct cttaagcggc cgcaagctag cttgccgcca
     1021 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1081 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1141 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1201 ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga
     1261 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1321 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1381 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1441 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1501 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1561 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1621 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1681 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1741 gagcctctag actccctcag cggcgcgcct ggtgttcctc gagatcggag tcctaaatcg
     1801 actgccttag cggtaggaca gtagatcagt cagcgacgtg acgaaccctc gcaggatcaa
     1861 gatcccacgt gtctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     1921 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1981 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2041 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2101 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag
     2161 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     2221 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
     2281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2881 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4201 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4261 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4321 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4381 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4441 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4501 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4561 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4621 aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc
     4681 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca
     4741 tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt
     4801 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc
     4861 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg
     4921 caggtgccag aacatttctc tatcgatagg taccgag
//
