LOCUS       sequence-258295-        5843 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 5843)
  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 5843)
  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_Axud1_enh_394_Citrine
FEATURES             Location/Qualifiers
     source          1..5843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..149
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          227..359
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     342..361
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      413..422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(419..421,422..424,425..1135)
                     /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(464..485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(725..744)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1072..1093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1309..1430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1320..1339)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1374..1393
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1678..1695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1849..2437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1929..1948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2608..3399,3400..3468))
                     /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     3231..3250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3469..3573)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3600..4055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3687..3706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3897..3918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4186..4234
                     /note="synthetic polyadenylation signal"
     misc_feature    4248..4339
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4288..4307
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag
       61 tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac
      121 accgagcgac cctgcagcga cccgcttaac agcgtcaaca gcgtgccgca gatcccaagc
      181 ttgcggccgc ttaagctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt
      241 tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt
      301 gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg
      361 tgtccactcc caggttcaat acagctctta agcggccgca agctagcttg ccgccaccat
      421 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      481 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      541 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      601 cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca
      661 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      721 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      781 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      841 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
      901 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
      961 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1021 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1081 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgagc
     1141 ctctagactc cctcagcggc gcgccgttgc gttccaaacg tcgtggagct ccctgtcaag
     1201 acacgaaccg ctaacgccac cttagtccgt gaagcatgat acgcgacata ccccaggaga
     1261 tctcacgtgt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     1321 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1381 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1441 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1501 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
     1561 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     1621 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     1681 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1741 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1801 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2041 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2281 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2641 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2701 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2761 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2821 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2881 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2941 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3001 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3061 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3121 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3181 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3241 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3301 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3361 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3421 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3961 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4021 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     4081 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     4141 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
     4201 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
     4261 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
     4321 ggtgccagaa catttctcta tcgataggta ccgagctcga gttgccaggc aaacagagcc
     4381 cgagagagga aaatgaagct ctgcctattt tacaggcaga tttttgcaac agtgcagatg
     4441 tcagccttgc tgtttcagaa cagccatgcc ctgccctata aacgtgtctc agatacatca
     4501 cattgttgtg ggtctgggaa actgggagtt gctggtcctt gcctaccctc tctaccaccg
     4561 ttaaagcaaa tcccatcctc agaaaagagg gaattcttag aaacctttgc ctttcaccag
     4621 caacatcctg caaactcgaa cgggttaatc ccctttagcc agcttaccca taatgaaagg
     4681 tttcactctt tgtcacctaa tggcagccag gtcttgtgtc taaaatacaa gcctagctgc
     4741 ctttgcatga aatgttttta atcatcgtgg aataaaaacc ttgcctggct agtgccaacc
     4801 aaagagatgt ttgtctgctc agcagatttt atcaaagctt ccttgatgtc tctgcctttg
     4861 taatgctaag cacttcactg tttggcatat ttaaagtagg atgcggtaca tgtgaagcat
     4921 ttcacagagc gatgatgtat cgagtgtgct ttaggaagta atgaaagcag cagactcatg
     4981 atttttgagc ttttaattac ttattaagtg gcatgacata aaggctacgg gggagatttt
     5041 ttttcagcat ccaacttact gactttgaag tcagtgccta aatacccttt gtgcctttga
     5101 aactttccgt ctgctcacag gggaacgcgc tgccgctctg ggcttcgttt cacgctgacc
     5161 acatttgttc agggcagggg atgggagtct gctcagacac aggtctctgc cttcgaggtg
     5221 gtggataaaa cgaggaacag actaaagacg tgccttcgga caaaggatgg ggagagcctg
     5281 gagagagcgc agagcctgga gcaatgagtt ctgacctggc ctgaaaaatg ccagcactac
     5341 agggggcaaa taaccatagc agcccccggg gtgcttttaa tgagctgaat tgcagtcttg
     5401 agtccagcct tccatcactc acaagccgct gccacagctg gttttagcag atgtctcctc
     5461 cgacagaaag gatgcggcaa agcacagtgg ttgacctccc ctccatctcc tctgggtccg
     5521 ctccgaccgc tgtgactaca aataaaagat aaagtcagga aaggccaaaa agaagaaaag
     5581 attctcagat ttgtggttgt gtttattgga gtttcttttc tttttgtttg tttgtttctt
     5641 tgttttgctt tttttttttt ctgtgttttt ttcccttgtt tttgactggt ttttcatatg
     5701 caggttttat ggggagtgac tgttggctag cccgggctcg agatctgcga tctaagtaag
     5761 cttgctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa
     5821 tatatttgca tgtctttagt tct
//
