LOCUS       sequence-259327-        6428 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 6428)
  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 6428)
  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_Cdh6_enh_318_Cerulean
FEATURES             Location/Qualifiers
     source          1..6428
                     /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        2247..2392
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2470..2602
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2585..2604
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2656..2665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2662..2664,2665..2667,2668..3381)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2707..2728)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2968..2987)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3315..3336
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3552..3673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3563..3582)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3617..3636
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3921..3938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4092..4680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4172..4191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4851..5642,5643..5711))
                     /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     5474..5493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5712..5816)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5843..6298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5930..5949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6140..6161
                     /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 ctcgagttcc
      181 atgggcagca ttgatctcct tcgctaaact gagtatgttc ttaaaatcac aggtcatctg
      241 attttggtga ctcaccccat gtaatctagc caacaagatt caaaggaaca ttttctgatt
      301 tttgcactgc tgttctaaac atcaccttca gtgcccttct atatacaatt cttcaggaga
      361 ggtaggctta ctttagttgt tatgcatatt tatagtggtt ctctaatcat tgctataaac
      421 aatacactga ggccaacaaa ctctcttaca aaagcaaacc cagtaaaatt ggtgggatgt
      481 gcacttgatt tagggtttct tctgtcactg agtacctgct gactgaattc tttccacata
      541 ttgtaaatag tgcctcatta cttggagctg taatgaaaaa gccaatttcc accacaccca
      601 aaaataatac aagtattttt ttaaatttgt gagattggtt ttaaatcatg acttatcttt
      661 aaaataaagt aagaagttta tggctttttt aatctttctc ttattttata ttaccttggt
      721 atttattata tgctatatgg ctaggaactc agtaattatt tttcatttaa aggtgaggat
      781 cacacgtaaa tatatgagct gtggatttac agaaaaatca gataccatag gagaatataa
      841 atacagtgtc aatgtcatat ttttatatgt tttgtaaaga agccaaaata aattattttt
      901 gtttttcttc catgcataaa agatccacta tcacttgctg aattagatta ttttatatga
      961 tcattgtttc aatttaattt aaaggaagta ggtccagaaa aataaaagct ttactcactc
     1021 ataaggactt aatgcacacc attcctcata ggaagaacat ttcaatgttt ttcaagtgtt
     1081 gacacaggtt ttcctttcgt ttacccatcc acagctgttg ggtgtttttt tttttttttt
     1141 ttttttaatg aaggagcata ttttgctaga cgaaccttca tttctccctc tctcttcctt
     1201 tcttcccacg attaaaaatc aaccacattc ccagagcacc actatctatc gaaactgaaa
     1261 gtcttcaggg tattttggaa gcgggagact gcaaagtttg aatgcggtat tccactggca
     1321 agctttcaaa cttatttgca gatcaggacc tcttttgtaa acgtctcccc aggaggaagg
     1381 cagttagaca cctgccagta gcctacagtg gcaccatcct actgtgatgt taatgagatg
     1441 aatgagagca ccacggagag ctgcaataat tacagggtga agagcttgtc ttttcctggc
     1501 cctcctctcc gggatgcctg ttctataaag cagggtcttt tgtaatacta attacaaatg
     1561 ttaattaaac atttctttgg aattttgccc tccattaata ggtcagataa acagggaata
     1621 gctgtgacag gcacaagtca ttatctgtca catgagaaaa atacaattta tttgctccct
     1681 tcctttggtc tctccaaaga cataacttct gattttgttt cgatttatta agcaggcatt
     1741 aaaaccactc aagtatagct tttctacaca gaaataaaat cgaattaatt cctacattaa
     1801 tggaattaaa cgtgcttgta aggaccttcc caacacaact caacctgacc tgtgttggct
     1861 tgaccccgag ttttaagcat aatcatgaag ctgtctctaa agccacttga aacccctttc
     1921 aaatgaagat ttaaggaaat gttgatgaca gctgttgatg acagcacaaa atctagcttg
     1981 atgtcctatt ctaacacaag caacacacgg catttcaaaa tcatgcacta atcacgttta
     2041 cagcctagtc cctagaatta ggggagcgac cccagctcag ctcagagcta agcacaagta
     2101 cttggcaggc atcatcacaa ggaccgctag cccgggctcg agatctgcga tctaagtaag
     2161 cttgctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa
     2221 tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg
     2281 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta
     2341 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aacagcgtca
     2401 acagcgtgcc gcagatccca agcttgcggc cgcttaagct gcagaagttg gtcgtgaggc
     2461 actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     2521 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     2581 cactttgcct ttctctccac aggtgtccac tcccaggttc aatacagctc ttaagcggcc
     2641 gcaagctagc ttgccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2701 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2761 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2821 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcgcc
     2881 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2941 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3001 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3061 gacggcaaca tcctggggca caagctggag tacaacgcca tcagcgacaa cgtctatatc
     3121 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     3181 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3241 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     3301 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3361 atggacgagc tgtacaagtg agcctctaga ctccctcagc ggcgcgccgt tgacttgggg
     3421 ctcctcagac ttactaggtg gcgggacagg ccaagcccaa tttaggatcg gtgtaccgca
     3481 gagcgtgaag caccctatcc agatctcacg tgtctagagt cggggcggcc ggccgcttcg
     3541 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3601 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3661 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3721 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     3781 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     3841 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     3901 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3961 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5761 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5821 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5881 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5941 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6001 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6061 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6121 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6181 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6241 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc
     6301 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6361 ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga
     6421 gcggccgc
//
