LOCUS       sequence-259019-        5204 bp    DNA     circular SYN 15-APR-2025
DEFINITION  enhancer reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5204)
  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 5204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_Zfhx4_enh_316_Cerulean
FEATURES             Location/Qualifiers
     source          1..5204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..105
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     54..73
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        975..1120
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1198..1330
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1313..1332
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1384..1393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1390..1392,1393..1395,1396..2109)
                     /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(1435..1456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1696..1715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2043..2064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2279..2400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2290..2309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2344..2363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2648..2665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2899..2918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3578..4369,4370..4438))
                     /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     4201..4220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4439..4543)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4570..5025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4657..4676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4867..4888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5156..5204
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttc catggacttt ccctttgctc ctcaggatag aaaacaaaga gaaaaatcct
      181 atcctgacag tgaaggagag ctgatcagtc tcaggaggtt tcattgtaag agtttaaaaa
      241 tcagctggcc taatgtttga tataagtctt tgtggatgac aagaggaaca ggcgagaaaa
      301 tagtcaagtt gaacagtaca ctccagtttg tcttttgcaa acttttccat cccccctcct
      361 gtcctttcag tgtactgttt tagatgcata atatatttcg ttacatcctt ctattcaaac
      421 ttaattacct tctccccttt gtcgtgacat gggcggaaag tttttgccgt gtcaggagag
      481 gtcgtcacac attcttttgc agttctcttc attaattcct tcttgcagaa gcattttcta
      541 tcgtgctggg aaggtagaag ggtgtaatag aagagccctg aaacatcagc aggaagctct
      601 aagctggtta atgcagtata attgtctgga catagactgg tctttgtttg tgtggttcac
      661 ttcgaatcag ctgtaattaa ctccagagtg tttccagatt gccagcaaag aaaaattaca
      721 gcttttctgt tttaaattgg aaagctcaca aagacattaa ctatgcagat tgcttttact
      781 ttgatatatt tttatcccat aagaaaaagc ttatttggct attttttttt cctccgccca
      841 aaatgtgagg accgctagcc cgggctcgag atctgcgatc taagtaagct tgctgcttca
      901 tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg
      961 tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg aattcgaaca
     1021 cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag gtgacgcgtg
     1081 tggcctcgaa caccgagcga ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc
     1141 agatcccaag cttgcggccg cttaagctgc agaagttggt cgtgaggcac tgggcaggta
     1201 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     1261 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     1321 ctctccacag gtgtccactc ccaggttcaa tacagctctt aagcggccgc aagctagctt
     1381 gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1441 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1501 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1561 tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg ctaccccgac
     1621 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1681 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1741 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1801 ctggggcaca agctggagta caacgccatc agcgacaacg tctatatcac cgccgacaag
     1861 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg
     1921 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1981 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     2041 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2101 tacaagtgag cctctagact ccctcagcgg cgcgccagga catgagtact tcccgagaga
     2161 gcgatgtcaa ccgaatcctg acgggacttc gtaattcgga ccaggtgttc ggagcttgtg
     2221 aggagataga tctcacgtgt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     2281 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2341 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2401 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2461 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     2521 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     2581 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     2641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4561 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4621 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4681 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4741 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4801 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4861 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4921 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4981 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca
     5041 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca
     5101 agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa
     5161 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtg
//
