LOCUS       sequence-258293-        5229 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 5229)
  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 5229)
  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_Snai2_enh_332_Citrine
FEATURES             Location/Qualifiers
     source          1..5229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        479..624
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          702..834
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     817..836
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      888..897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(894..896,897..899,900..1610)
                     /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(939..960)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1200..1219)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1547..1568
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1782..1903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1793..1812)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1847..1866
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2151..2168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2322..2910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2402..2421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3081..3872,3873..3941))
                     /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     3704..3723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3942..4046)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4073..4528
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4160..4179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4370..4391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4659..4707
                     /note="synthetic polyadenylation signal"
     misc_feature    4721..4812
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4761..4780
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ctcacgaggc ttgtttgagg tgttttgcat ccataattca ggggttcata gatttttatg
       61 gccagaagaa accattagat catctggcct ccccttcact attataaaag acatacaatt
      121 gcattcattt actcctgttg ttaacccaat tacttgtgtt tcactgtctt ccaaaaaaga
      181 catccaaaga cttcaaaaga tggcaagccc acttctccct tggaagttta tcccaacagc
      241 taatcaccct aagtattaca caactaatgt aggcttggga ctaataaagc ttagtcccct
      301 atggatttgt gtttcatggt tgattgattt ccatgtctaa caagctgtga gctgttggct
      361 agcccgggct cgagatctgc gatctaagta agcttgctgc ttcatccccg tggcccgttg
      421 ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat
      481 gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg
      541 gcggcgcggt cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga
      601 gcgaccctgc agcgacccgc ttaacagcgt caacagcgtg ccgcagatcc caagcttgcg
      661 gccgcttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
      721 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
      781 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
      841 actcccaggt tcaatacagc tcttaagcgg ccgcaagcta gcttgccgcc accatggtga
      901 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      961 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1021 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1081 ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1141 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1201 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1261 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1321 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1381 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1441 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1501 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1561 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tgagcctcta
     1621 gactccctca gcggcgcgcc tggtgttcct cgagatcgga gtcctaaatc gactgcctta
     1681 gcggtaggac agtagatcag tcagcgacgt gacgaaccct cgcaggatca agatcccacg
     1741 tgtctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     1801 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1861 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1921 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1981 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
     2041 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     2101 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     4081 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4141 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4201 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4261 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4321 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4381 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4441 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4501 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     4561 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     4621 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
     4681 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     4741 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     4801 gaacatttct ctatcgatag gtaccgagct cgagttgcca ggggtccact gattccacat
     4861 ccatttagta aagtagaaat aagcccattg taacaggaga ttgccatagg atctgtatca
     4921 ccagctttgc aaatgaacat gccttgatgt ttaattttcc gtgagttcct ctgcatttag
     4981 gtaccagcag gggaccatta tactgtgaca gcactcagtt ctgatgacat tatcttaaac
     5041 ttttccctgt tgtccgtcct cacccctgtt gtgctgccgc catctattaa cattccactg
     5101 acaacaatga aagatgttag agtagaacat gttaatgcag tagccaagct aggatgcttt
     5161 cacctggatg gatacaaaag tttggcagca gctttatagt taattagcta gatcacatca
     5221 tgccaattc
//
