LOCUS       sequence-258391-        7349 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 7349)
  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 7349)
  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_334_Citrine
FEATURES             Location/Qualifiers
     source          1..7349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2385..2530
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2608..2740
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2723..2742
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2794..2803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2800..2802,2803..2805,2806..3516)
                     /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(2845..2866)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3106..3125)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3453..3474
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3690..3811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3701..3720)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3755..3774
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4059..4076)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4230..4818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4310..4329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4989..5780,5781..5849))
                     /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     5612..5631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5850..5954)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5981..6436
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6068..6087)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6278..6299
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6567..6615
                     /note="synthetic polyadenylation signal"
     misc_feature    6629..6720
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6669..6688
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atgacaatgg cattgctctt tggcaaggct ctcggcttca aatctgttct catgaaactt
       61 tcagctgagg tcttgcagac tcagactttt agttactgtg gtttcaagac gcacctcagc
      121 ctattgatgc tcagctaaaa aagccaccaa ttccaatcct ctggtagctt acctatcatc
      181 actagtgtaa accaaccagc ctttcctcca caagaagcat aaattccctg gagccctgct
      241 tccagatggg gaaattgcct tcagtgacac ctgttacatc tcatatcaga tagcacaacg
      301 tctgtaattc tgtcatggac tcatcctgca actttgtgag tcacactgca tgtttcatgc
      361 accaagcgaa ctaaagcaaa caaacaggaa ctatttgtat tcttcttgtt tatgccagga
      421 cttataataa atcccttcag aaacacttcc gaacaggtag cagccagcac acaagtttgg
      481 ccaaggtttt caactagtct ttacctgttg cagccgttgg gaaatttcta tctgttggtg
      541 gaaatgtact tgtacctgca tgctggcact gggagtgtga gcaggtgttt gcaaaccgca
      601 gttatattgc taaccatgct tactgtctca gactcaaaaa tattaatcat gatggcaatg
      661 tctggtaccc aaaatacctt gctgcaaaaa gttctcattc aatttgtctt accgcagtga
      721 tcagaactgc tctaagcaat aaatgcctct cgagctaatt tctgtatgca gcgcaacgct
      781 acttaccccg aaggcccttt aagatttgac ccctttcact gtttttcctg ctgtcaggca
      841 ggacagcaaa agaggtggta tgcgggctga aaagccaagc agccctcaat cagccctttg
      901 tctttcatga ccagcttttt atccaggggc aacagtcttt ttccattacc agtaaaaatg
      961 aaccccccag taggggtgat gaagaaacaa taaaactaat cattaagcag gcctgctgag
     1021 aagcgaattc cagtcccagg tcagatctaa ttgcattctg acactgaagc agtggcctgg
     1081 tcatttatat tagcgatgtc atgttccttg ctgtttattt ctcaacgttt tgctccctgt
     1141 tcccattaac aaataaatgc acgtaaagat caaaatacac atactgccct gtggtgtact
     1201 ttgcaaattt ttaaaatacc gtctcttttt atccaagatc aattgtggaa aaactaaaaa
     1261 caaatataac attttggaaa ataatgagaa actgtaaaag ggatagtcag taaaaattat
     1321 ggtagtatta tggcctatga ccaaggaaaa ttaaatactc aggggaggaa agacaaattt
     1381 aaccatgcgt atgaacacac aagataaaca tatgtggctg tgattttcaa aaccagagat
     1441 attcagatag ctatctttga gtaattacaa cagcaataca acacttctga gctctgtaca
     1501 gcacatgcat gcacgtggca tgcactcgca tccctgaatg cattcctttc aatacaaaca
     1561 tccttggtct ctttcctctt ttgccacttc tcttttactg cgttcatcat tactgatgtt
     1621 ttgcccctat ctgcatcccc agtctcctct cccactgttt tttgcccaga ctcccagcca
     1681 gcttcccagc ctgagactct tccttcgaga atctaaacaa gatgcctatg catctgactg
     1741 aacacaatgg gcccctcttc aaagccaagc aatatttttc ctatcattgt ttgcagggga
     1801 agagtcactc tacgttctgc tcagatatta cggttacgct agccttatta gctcatgggt
     1861 atactttgct gttacaagag ggtttgtaca actgacaagg ttgcttaatc ccttcctagg
     1921 tccctcagca tttgtaacaa gttttgtaat tgctttttaa ataaaatgcc ctgaccctgg
     1981 aaactcattt gtgagccagc tcagttctct ggcttttttc ccttcatata gcttttatga
     2041 cctctcctat ctggatatcg ggccctttct cagaagtatc ttagtccaat cctcttgatg
     2101 ctatccagag ctaaaagccc tcagcatgtg ctttggatcc gatccctacc tggctgctct
     2161 gtccagaggg gtgttgatca aatctggtaa caacaaatca gggaccagaa aatgcatttg
     2221 tagaatctgg agtttttcca tctaattgca agcaggcctg ttggctagcc cgggctcgag
     2281 atctgcgatc taagtaagct tgctgcttca tccccgtggc ccgttgctcg cgtttgctgg
     2341 cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc
     2401 ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca
     2461 ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg
     2521 acccgcttaa cagcgtcaac agcgtgccgc agatcccaag cttgcggccg cttaagctgc
     2581 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
     2641 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     2701 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccaggttcaa
     2761 tacagctctt aagcggccgc aagctagctt gccgccacca tggtgagcaa gggcgaggag
     2821 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2881 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2941 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     3001 ggcctgatgt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3061 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3121 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3181 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3241 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3301 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3361 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     3421 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3481 gccgggatca ctctcggcat ggacgagctg tacaagtgag cctctagact ccctcagcgg
     3541 cgcgccgttg cgttccaaac gtcgtggagc tccctgtcaa gacacgaacc gctaacgcca
     3601 ccttagtccg tgaagcatga tacgcgacat accccaggag atctcacgtg tctagagtcg
     3661 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3721 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3781 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3841 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3901 taaaatcgat aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     3961 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     4021 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     4081 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4141 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4741 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4801 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4861 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4921 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4981 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5041 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5101 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5161 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5221 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5281 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5341 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5401 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5461 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5521 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5581 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5641 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5701 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5761 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5821 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5881 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5941 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     6001 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6061 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6121 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6181 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6241 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6301 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6361 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6421 ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6481 gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag
     6541 gtacgggagg tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg
     6601 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     6661 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
     6721 atcgataggt accgagctcg agttgccagg gtgccattgt gtgcatggtt tgtgttccag
     6781 ctgacactga tatttactta taaaagcaga acctatatag atcaggaata cttggggcat
     6841 aatttctcat ttggaataga tcacgagagc aacttacaca gctctcaggt ggatacatac
     6901 actgtggaca ttgctgtgtc ttcaacttct ttggatttca gatcctgcgc ttcaaattca
     6961 cttcagaatc actcaaatgt aaaagttatt ggcatctcca gtacatctga gagagcagtt
     7021 ctgaggttca cagctgacca caatttcttt acattcacac cgggtaattt taactatctc
     7081 ttgacttgct ctgttgcaga ttaaattata aaaaaaaaaa aaaaaaagaa tgttcattat
     7141 ttaagcgatt tattataatg ctaaacatta tagattagtt caacaattga tgtgatcagt
     7201 tttgctgcac tgcagtcctt gaataaacac agatttacac cagtgggcag tttggccttc
     7261 aaaattgttt tgtatatgcg gcatcaagat tttttaaaag ctttacttac tttatttcag
     7321 tctggctgaa ctggtgtggt gaggaagga
//
