LOCUS       sequence-258291-        6895 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 6895)
  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 6895)
  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_Snai2_enh_242_Citrine
FEATURES             Location/Qualifiers
     source          1..6895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2210..2355
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2433..2565
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2548..2567
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2619..2628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2625..2627,2628..2630,2631..3341)
                     /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(2670..2691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2931..2950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3278..3299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3513..3634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3524..3543)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3578..3597
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3882..3899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4053..4641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4133..4152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4812..5603,5604..5672))
                     /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     5435..5454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5673..5777)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5804..6259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5891..5910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6101..6122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6390..6438
                     /note="synthetic polyadenylation signal"
     misc_feature    6452..6543
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6492..6511
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aatcaggact cagctagtca agaaaacaca ttctgcacaa aaaatctctt ctcagtgctg
       61 aaggcacagt ctgaatggac acagagtctc agctacactt taattgtcct gagactatcc
      121 tttcaggcat gtggctggag caggtcacac tgctgtagct gctgtcctca ggcagtaaga
      181 cacagtgaag acagggcttt acaaagacct ttttttttaa agagcttttt tttttttttt
      241 tctttttcag gactagtcac agtaaataga gtggttattt ggtatgtgag ataagaattc
      301 gccaacatca tttgaacaca ctagttttct ttcagtgttt tgtcacttga ccagtttaat
      361 gctaaggtta aaaagctctt acacaacatg tatccaccct cttgtgtgtc aaaaatgtga
      421 ggaatgtttg caagaaacag cagcttgcgt acaattcctg tcaccaaagc ccaaagttga
      481 aatggctaac gtttcctgga tctgcttagc tcacctatgg agacagagct tgccgtgcct
      541 gtgtgcaaac agaagcagag tcattaaaac gggagcagtt cttgggcaag ctccagtttg
      601 catgttacat attcataaat agctgtgtta gaatggcttc caacagtgca aacgtcaatg
      661 gagcctgctg gagcaagcct taggtatgcc atctatgtgt ctgattgcct ggagttcaga
      721 ggaacttgct gaatgtatta ccaacaagaa tgaaaaggct tgcatcaggg tagctggcca
      781 ggtacaattg ctgggtattt aaaaggacgt atcagtacca aattgtttca cagctgtaat
      841 ttttaaactc acttgctgtt tgtgtaatgc ttgagaatct ccaacaggaa tatttcattc
      901 tgttgttctt tcaaagagca catattaatt tttgttgtaa caaaaaattt tcattagggc
      961 ttggctaata gcactgatta tttgtttatt ttcactctgt gcaacctatt taattgtgta
     1021 tacacatggc tataattacc actttgaaat gcaaatacac agttactaag gataaccctt
     1081 cttgttttca agtagctaac actccttttt cataaatgct aaccaaatca taagatccca
     1141 gtaaaacagc acagtttaag ctctccccac attgcaatgg ctgcctcctg ctccaaatta
     1201 ctctctctgc cactgtcata gcttccttca atttatcacg tcaggggccc taggcaaggg
     1261 ccacgaggtc atttgaccaa agtactggcc agcttacagc tgagtgtaca ccaggactcc
     1321 tctaagtgat aggaagaaga gtgagtgcgt tttatgccaa actgtgtctc atccagtctc
     1381 caactccatt tgtaacctat ccacggaaag gtctgtgttg gtacagagtc atttgttact
     1441 ctcagttatt caccagatag tttattgcta tttatttcca cccacagact gctctcagac
     1501 tttcctctgg gacaggattt gcttcctaaa gggaggattt agggatttga ctaagcagca
     1561 tgagcagcag aagatatcac ataacaatgg ctttacttcc taaatgtcaa actcacaaag
     1621 tgctttcaaa gtgaccgtac caacccacct tccttttcgc atcacttccc cttgccccca
     1681 atcaattacc gatttcagtg gttacccata gaagtttctc tttcagcagc atttttgtaa
     1741 gcaaatattt gcttgtccat ctattcacag agagcacatt aaagcagcgt gccagttaga
     1801 gtgtgggctg tctttgaaca gatgttatga atactgtttt gcaggatttg cccagcagta
     1861 gatgagaaca agtaatgagg aaactaggaa catgggtgac gaggccagaa ggtgtggaag
     1921 ctttccagga gcagtgggag ctgggtggag aaaacttgac ttactgatat gcagccacga
     1981 gcccttctca gactttattt gttcttctta atggtcactc tcaagaaaag ttattcatca
     2041 gggctcattc ccttgagaat taattttcct gtgccttcga ggctgttggc tagcccgggc
     2101 tcgagatctg cgatctaagt aagcttgctg cttcatcccc gtggcccgtt gctcgcgttt
     2161 gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac
     2221 cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg
     2281 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg
     2341 cagcgacccg cttaacagcg tcaacagcgt gccgcagatc ccaagcttgc ggccgcttaa
     2401 gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
     2461 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     2521 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagg
     2581 ttcaatacag ctcttaagcg gccgcaagct agcttgccgc caccatggtg agcaagggcg
     2641 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2701 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2761 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     2821 gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2881 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2941 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3001 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3061 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3121 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3181 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3241 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3301 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgagcctct agactccctc
     3361 agcggcgcgc ctggtgttcc tcgagatcgg agtcctaaat cgactgcctt agcggtagga
     3421 cagtagatca gtcagcgacg tgacgaaccc tcgcaggatc aagatcccac gtgtctagag
     3481 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3541 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3601 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3661 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3721 tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     3781 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     3841 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     3901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4741 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4801 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4861 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4921 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4981 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5041 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5101 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5161 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5221 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5281 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5341 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5401 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5461 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5521 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5581 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5641 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5701 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5761 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     5821 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5881 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5941 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6001 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6061 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6121 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6181 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6241 atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6301 tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt aagtaatatt
     6361 aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca ttacatctgt
     6421 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     6481 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     6541 tctatcgata ggtaccgagc tcgagttgcc aggcacttag gtagatgcag gtccagatat
     6601 ccatagcgca ttaagagatg cctttagcca gctgaaaact ctttttgatc tctctgataa
     6661 taccaggcaa gcactgatct ggaaaggcag attaccacca aggattaagc agtacaatag
     6721 catgctgaga atgtaagaga atgatgaggt ttagcaaaca agcgtggaac atatttgaac
     6781 tcaacatcga acaccaaaag aagacatagg aagagaactg aggctgtagg gaactgccct
     6841 gaaaaatgtt atatgagaga gggcaaaact gtctaactac tagattttaa cttaa
//
