LOCUS       sequence-259016-        5396 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 5396)
  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 5396)
  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_17_Citrine
FEATURES             Location/Qualifiers
     source          1..5396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..105
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     54..73
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1165..1310
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1388..1520
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1503..1522
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1574..1583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1580..1582,1583..1585,1586..2296)
                     /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(1625..1646)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1886..1905)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2233..2254
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2471..2592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2482..2501)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2536..2555
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2840..2857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3011..3599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3091..3110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3770..4561,4562..4630))
                     /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     4393..4412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4631..4735)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4762..5217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4849..4868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5059..5080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5348..5396
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccagggctgg agttatggct gtgggatctc atctgactaa aagaatagcg
      181 tgtacacagt aatcagctct gttataattc tgccgtggtg tatggaaagg aaattgcagt
      241 agcattattc taggagtagt ctacagactt tgtctttttt ggtacgtctt gaaagggagc
      301 cagtgtctcc ccagcttgag tcaaccatac cctatctaag taaagtggac tgagatcatc
      361 tagatgctgg tcttcatttc taatgagcct tcccagtaaa acaggaaatg ataatcttgg
      421 aagggggaaa aaaaaaatgg tcaaaaatca gccagggaac atttaaaggt ttgttctgta
      481 ctagcagagc taagaaaaag gcctgagaaa agtgtgtcct gggatcactc acagttaaag
      541 cactgattaa ttaagcagag ggtcattctt cagaaagact ggagggcagt ttgggcccag
      601 gggctgtaat ttggacactt ctggagttct agatacactg agctattctg cgctgggcct
      661 gtgaatatgt gcattctttt ggctgaacaa tggcatcttt attcaaatgt gagccactaa
      721 gtaagtggct gtctcctaat atgggtggta tttcttcctg ctctgcctca atctccgctg
      781 ctttcccgtt gccttatcag tgctttgtag cacaactgac aaaacggaac ctattcacaa
      841 tgctaaaaac ctcccctttc agagacacag aaaaaacgcc ccaaatgctt acatatcctc
      901 caattctgtg gcagggtgtt tacttcttta aatttggttc tgcttatctg acactgtttt
      961 ggaaattaat aaagtggtgc ctggctttta ttgttttgac tatgggaagg aaattaaccc
     1021 acaactgtta caagtgtctg ttggctagcc cgggctcgag atctgcgatc taagtaagct
     1081 tgctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata
     1141 tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg
     1201 aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag
     1261 gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa cagcgtcaac
     1321 agcgtgccgc agatcccaag cttgcggccg cttaagctgc agaagttggt cgtgaggcac
     1381 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
     1441 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
     1501 ctttgccttt ctctccacag gtgtccactc ccaggttcaa tacagctctt aagcggccgc
     1561 aagctagctt gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1621 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1681 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1741 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt gcttcgcccg
     1801 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1861 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1921 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1981 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2041 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2101 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2161 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     2221 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2281 ggacgagctg tacaagtgag cctctagact ccctcagcgg cgcgccttgc agtaattcac
     2341 gaggccaaag gcgaaccatt tccaccagcg gtactccggc tcattcttac cgcgcagatc
     2401 gttgtggaca gatcgcgaaa gatctcacgt gtctagagtc ggggcggccg gccgcttcga
     2461 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2521 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2581 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2641 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     2701 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2761 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     2821 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3781 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3841 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3901 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3961 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4021 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4081 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4141 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4201 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4261 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4321 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4381 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4441 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4501 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4561 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4621 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4681 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4741 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5161 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc
     5221 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5281 tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag
     5341 cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtg
//
