LOCUS       sequence-249940-        6905 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 6905)
  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 6905)
  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_Ets1_enh_117_Citrine
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2726..2871
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2949..3081
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3064..3083
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3135..3144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3141..3143,3144..3146,3147..3857)
                     /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(3186..3207)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3447..3466)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3794..3815
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4029..4150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4040..4059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4094..4113
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4398..4415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4569..5157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4649..4668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5328..6119,6120..6188))
                     /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     5951..5970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6189..6293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6320..6775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6407..6426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6617..6638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttgc
      181 caggtgctcg ctgttaatgc ctctgctccc tcaggactag gccaagtgat ggggttaatt
      241 tgggaggggg ctggaggtgc agcttcagga gatggcaatt aacaggagac agcgctcagg
      301 gaactcactg caaatgaaaa aacaacaggt cctcgctcct gtggtgttaa ggagtggctt
      361 aagggggttg tccggagccc agaaatctgg gagtaatttg ggacaaaaca atgtcctctg
      421 caccactttg atgctgcaga aacatgagaa catgcagaga tagtgtcgca tctcagagcc
      481 tccccacccc aaggccgtgg agatttagat ttagattctg atcagggcct gcagtgtgct
      541 gcaatcaata cagccaaggc tgaaggcgat ttaattgtgg caagtaatga cccaacaccc
      601 tggggagatg cttgtggatg gaagtcgagg aatttgctca gatctgtgca gaggtgggca
      661 atgggatgag cccaagcttg gagtttttac ccttttttgg aagaaaaatg tctctccacc
      721 tctgtttttt ccccttaact caaaaatcat catctattct ccgtgagaga atgggcagct
      781 gaatatcacg aatggcataa ttccaggaaa agctgctccc ttttatactg cagagaggaa
      841 tgtggaaaag caattcaaaa agcccaagag aaataattat caattttcag ccacagcagc
      901 ctggagccaa tggggcacac atggcagagc tcagcttagg gctgccatgg ctgaacgctt
      961 gtgccaatag agcaagtgta ggtgggtgcc aatacagctg aagtacacat cataaatagg
     1021 tgtaagcgtc ctgacggttg cagaagtgca tgaacacttg gtgttacaag gttgtgaact
     1081 tctgggagtc tcatagtgaa ataccaaata accactggtg ccttttaatt ttggctcttc
     1141 tctgactatt tactcttgct gggttttttt ttttagctat aaccttctgg agggtgtggg
     1201 cgatgctgtt tttaaggtga ccgcggagga tggctccagc atcactcaaa agaactttct
     1261 gggccctcta gtggtgtgtg ctgtgctgag gacctgatct ccctatcctt ttgttccacc
     1321 aaaataaaaa ctagtcatta ttttccccat tatttttcac aaaatgagga aaatcccaaa
     1381 tctccctggt cttatttttt atttttattc ttcttagaac gcacctatgg aaggagcagt
     1441 gcatccgttg ggttaaaacc ctccctctgc ctcttcagaa ccaacgaacg tatgcagaca
     1501 ttcatttcaa aactgaaaac ccctctcgaa aagacaaagg tgggctgata tgttttgaag
     1561 acatcagggc aatgggggac tcatgccttt ctatcatttg tttctcctgt tatggcttgt
     1621 gaagaaaata atgtcacagg ggtcttatca gggagctggg gaaggacagg ggatttggga
     1681 agagcttacg tatttctttc caagctgtgt ttctaaaata agttgatgtt tatttggttg
     1741 cgagggaaaa acgtttaatt tgatatcctg tccgtgctgg aaacagcaaa cattttaaca
     1801 aaggggagga tgggaaaagg tggctggaaa agcagcgtct gtaggtgttt gggctttgtt
     1861 tgttgagtaa caccattgct gacagcatca tccccagcca ccctgcctgc gtacaccagc
     1921 acttatcccc agggcgcaca cagctttgtg tcctgcctac cccacacaac tatttgtgct
     1981 ctgccaggta ggggtgtgag ttttgactga aatggctctt tctggtggtt tagcagatgt
     2041 aggcagaggc agcatgagct gcaaaggcct tggggtgggt tctgcccttt actacctggg
     2101 catccagcct ttcttttgga tttttgacac cttcataggc ttgctcaaga ggtaaattaa
     2161 gggtgttaag atggatatcc agggatcttg ggatagattt aggggaaacc tccccttcat
     2221 attgagttcc cagcttaagc aatgctcttc cacatctttc cctgttggcc tgcatgggat
     2281 ggtgcagggc gcagccatgt tgaggtgtcc ccttgggttc aagactgaat ttttccaagt
     2341 gtggacgctg cagcagactg ctctgaaggg ctcagcataa gggatcccaa aaccttcacc
     2401 ttgccaaccc agcctgctgg ttttagtggt ttgcaaatgc aaaccagctt gactgtgctg
     2461 ctttggcagc cgtaggagct tcacatcacc atcctgattg catttggatt tgggcactgt
     2521 agtcctggaa aacacttgga gcagccaaca tcaaatgtaa caaagcaaga cctggatgtg
     2581 tgaggatgga gtgatgccct gttggctagc ccgggctcga gatctgcgat ctaagtaagc
     2641 ttgctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat
     2701 atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc
     2761 gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa
     2821 ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa
     2881 cagcgtgccg cagatcccaa gcttgcggcc gcttaagctg cagaagttgg tcgtgaggca
     2941 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     3001 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     3061 actttgcctt tctctccaca ggtgtccact cccaggttca atacagctct taagcggccg
     3121 caagctagct tgccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3181 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3241 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3301 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc
     3361 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3421 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3481 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3541 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3601 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3661 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3721 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     3781 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3841 tggacgagct gtacaagtga gcctctagac tccctcagcg gcgcgcctgg tgttcctcga
     3901 gatcggagtc ctaaatcgac tgccttagcg gtaggacagt agatcagtca gcgacgtgac
     3961 gaaccctcgc aggatcaaga tctcacgtgt ctagagtcgg ggcggccggc cgcttcgagc
     4021 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4081 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4141 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4201 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     4261 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     4321 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     4381 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4741 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     4801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4981 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5581 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5641 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5701 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5761 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5821 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5881 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5941 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6001 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6061 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6301 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6361 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6421 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6481 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6541 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6601 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6661 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6721 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     6781 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6841 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     6901 gccgc
//
