LOCUS       sequence-259034-        5264 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 5264)
  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 5264)
  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_Msx1_enh_389_Cerulean
FEATURES             Location/Qualifiers
     source          1..5264
                     /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        1028..1173
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1251..1383
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1366..1385
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1437..1446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1443..1445,1446..1448,1449..2162)
                     /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(1488..1509)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1749..1768)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2096..2117
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2339..2460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2350..2369)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2404..2423
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2708..2725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2879..3467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2959..2978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3638..4429,4430..4498))
                     /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     4261..4280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4499..4603)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4630..5085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4717..4736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4927..4948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5216..5264
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttc catggcccac tagaccatgt tcccccagtg ccacatctat ctttttcttg
      181 atcacctcca aagatggtga ctgcaattat actagtttct gcctttggat atgagtgctg
      241 attttagtta caactagcca ggtatatgta attaagctca gattggactg cagtgctacc
      301 tacagtaaga catgtgggag gcagtgggag caggaatgct gggagtactg agactttgca
      361 aaacagagct gaaaattgaa tcccatttgt gcctactcca gaatctatct ggtattgtaa
      421 catatccttt aagactgcta aaggattgat cccaaactgg gaggtgtctt tccatctacc
      481 tctgtggtct tttttacttg gaaatctttt gttatcttgc agtaaatgct ctggtataag
      541 ctgaagttcc aaccctcctt ttcttgcctc tggtgatcct atagagactg ctcaaattac
      601 ccaagaggtt tgagatagcg tattgttttc atggacttaa ctctgtgtaa ttatgcatga
      661 gtaatcctcg tttatttctg ttatccttta attgagctat aagatttttt aagcttaaaa
      721 aataccacag gctttctgag ggctctaaat taaaatgcaa caaaacaaaa aagtgcactc
      781 cttagaccaa gaatctgcca tattttattg taaaagaagt ggcttttgtg ttgaatcctt
      841 cagaacaaca gtgtggagtc tttttctttt attttcaatt gtcctgaagt gctgtgtctc
      901 aggaccgcta gcccgggctc gagatctgcg atctaagtaa gcttgctgct tcatccccgt
      961 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
     1021 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat
     1081 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
     1141 gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc
     1201 aagcttgcgg ccgcttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
     1261 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     1321 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     1381 caggtgtcca ctcccaggtt caatacagct cttaagcggc cgcaagctag cttgccgcca
     1441 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1621 ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
     1681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1861 acaagctgga gtacaacgcc atcagcgaca acgtctatat caccgccgac aagcagaaga
     1921 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2041 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2101 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2161 gagcctctag actccctcag cggcgcgcct cgctaagaga actccggtac gacgcgatgc
     2221 gactacagca gggaccttca agcgctcttc tcgggagaat cgctagtcgc gcttggataa
     2281 ccttccgaga tctcacgtgt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     2341 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2401 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2461 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2521 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     2581 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     2641 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     2701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4621 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4681 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4921 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4981 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5041 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca
     5101 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca
     5161 agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa
     5221 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtg
//
