LOCUS       sequence-249909-        7345 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 7345)
  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 7345)
  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..7345
                     /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        3168..3313
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3391..3523
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3506..3525
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3577..3586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3583..3585,3586..3588,3589..4299)
                     /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(3628..3649)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3889..3908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4236..4257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4469..4590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4480..4499)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4534..4553
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4838..4855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5009..5597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5089..5108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5768..6559,6560..6628))
                     /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     6391..6410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6629..6733)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6760..7215
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6847..6866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7057..7078
                     /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 cagggtgcca ttgtgtgcat ggtttgtgtt ccagctgaca ctgatattta cttataaaag
      241 cagaacctat acagatcagg aatacttggg gcataatttc tcatttggaa tagatcacga
      301 gagcaactta cacagctctc aggtggatac atacactgtg gacattgctg tgtcttcaac
      361 ttctttggat ttcagatcct acgcttcaaa ttcacttcag aatcactcaa atgtaaaagt
      421 tattggcatt tccagtccat ctgagagagc ttttctgagg ttcacagctg accacaattt
      481 ctttacattc acaccgggta attttaacta tctcttgact tgctctgttg cagattaaat
      541 tataaaaaaa aaaaaaaaaa agaatgttca ttatttaagc gatttattat aatgctaaac
      601 attatagatt agttcaacaa ttgatgtgat cagttttgct gcactgcagt ccttgaataa
      661 acacagattt acaccagtgg gcagtttggc cttcaaaatt gttttgtata tgcggcatca
      721 agatttttta aaagctttac ttactttatt tcagtctggc tgaactggtg tggtgaggaa
      781 ggaatgacaa tggcattgct ctttggcaag gctctcggct tcaaatctgt tctcatgaaa
      841 ctttcagctg agttcttgca gactcagact tttagttact gtggtttcaa gacgcacctc
      901 agcctattga tgctcagcta aaaaagccac caattccaat cctctggtag cttacctatc
      961 atcactagtg taaaccaacc agcctttcct ccacaagaag cataaattcc ctggagccct
     1021 gcttccagat ggggaaattg ccttcagtga cacctgttac atctcatatc agatagcaca
     1081 acgtctgtaa ttctgtcatg gactcatcct gcaactttgt gagtcacact gcatgtttca
     1141 tgcaccaagc gaactaaagc aaacaaacag gaactatttg tattcttctt gtttatgcca
     1201 ggacttataa taaatccctt cagaaacact tccgaacagg tagcagccag cacacaagtt
     1261 tggccaaggt tttcaactag tctttacctg ttgcagccgt tgggaaattt ctatctgttg
     1321 gtggaaatgt acttgtacct gcatgctggc actgggagtg tgagcaggtg tttgcaaacc
     1381 gcagttatat tgctaaccat gcttactgtc tcagactcaa aaatattaat catgatggca
     1441 atgtctggta cccaaaatac cttgctgcaa aaagttctca ttcaatttgt cttaccgcag
     1501 tgatcagaac tgctctaagc aataaatgcc tctcgagcta atttctgtat gcagcgcaac
     1561 gctacttacc ccgaaggccc tttaagattt gacccctttc actgtttttc ctgctgtcag
     1621 gcaggacagc aaaagaggtg gtatgcgggc tgaaaagcca agcagtcctc aatcagccct
     1681 ttgtctttca tgaccagctt tttatccagg ggcaacagtc tttttccatt accagtaaaa
     1741 atgaaccccc cagtaggggt gatgaagaaa caataaaact aatcattaag caggcctgct
     1801 gagaagcgaa ttccagtccc aggtcagatc taattgcatt ctgacactga agcaatggcc
     1861 tggtcattta tattagcgat gtcatgttct ttgctgttta tttctcaacg ttttgctccc
     1921 tgttcccatt aacaaataaa tgcacgtaaa gatcaaaata ctcatactgc cctgtggtgt
     1981 actttgcaaa tttttaaaat accgtctctt tttatccaag atcaattgtg gaaaaactaa
     2041 aaacaaatat aacattttgg aaaataatga gaaactgtaa aagggatagt cagtaaaaat
     2101 tatggtagta ttatggccta tgaccaagga aaattaaata ctcaggggag gaaagacaaa
     2161 tttaaccatg cgtatgaaca cacaagataa atatatgtgg ctgtgatttt caaaaccaga
     2221 gatattcaga tagctatctt tgagtaatta caacagcaat acaacacttc tgagctctgt
     2281 acagcacatg catgcacgtg gcatgcactc gcatccctga atgcattctt ttcaatacaa
     2341 acatccttgg tctctttcct cctttgccac ttctctttta ctgcgttcat cattactgat
     2401 gttttgcccc tatctgcatc cctagtctcc tctcccactg ttttttgccc agactcccag
     2461 ccagcttccc agcctgagac tcttccttcg agaatctaaa caagatgcct atgcatctga
     2521 ctgaacacaa tgggcccctc ttcaaagcca agcaatattt ttcctatcgt tgtttgcagg
     2581 ggaagagtca ctctacgttc tgctcagata ttacggttac gctagcctta ttagctcatg
     2641 ggtatacttt gctgttacaa gagggtttgt acaactgaca agattgctta atcccttcct
     2701 aggtccctca gcatttgtaa caagttttgt aattgctttt taaataaaat gccctgaccc
     2761 tggaaactca tttgtgagcc agctcagttc tctggctttt ttcccttcat atagctttta
     2821 tgacctctcc tatctggata tcgggccctt tctcagaagt atcttagtcc aatcctcttg
     2881 atgctatcca gagctaaaag ccctcagcat gtgctttgga tccgatccct acctggctgc
     2941 tctgtccaga ggggtgttga tcaaatctgg taacaacaaa tcagggacca gaaaatgcat
     3001 ttgtagaatc tggagttttt ccatctaatt gcaagcaggc ctgttggcta gcccgggctc
     3061 gagatctgcg atctaagtaa gcttgctgct tcatccccgt ggcccgttgc tcgcgtttgc
     3121 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
     3181 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
     3241 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     3301 gcgacccgct taacagcgtc aacagcgtgc cgcagatccc aagcttgcgg ccgcttaagc
     3361 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
     3421 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     3481 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtt
     3541 caatacagct cttaagcggc cgcaagctag cttgccgcca ccatggtgag caagggcgag
     3601 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3661 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3721 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     3781 tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     3841 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3901 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3961 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4021 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4081 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4141 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     4201 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4261 gccgccggga tcactctcgg catggacgag ctgtacaagt gagcctctag actccctcag
     4321 cggcgcgcct ggatctgccg acaaccaggt tcgagcaagg acgcaggttg tgttgccggt
     4381 aagtcgacaa tcgagcttgt gatcggtcct tgaagccaga tctcacgtgt ctagagtcgg
     4441 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     4501 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4561 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4621 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4681 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     4741 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     4801 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4861 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4921 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4981 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5041 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5101 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5161 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     5221 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5281 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5341 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5401 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5461 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5521 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6721 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     6781 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6841 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6901 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6961 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7021 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7081 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     7141 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7201 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     7261 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     7321 tacgggaggt acttggagcg gccgc
//
