LOCUS       sequence-458191-        4316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A gateway compatible 5' entry clone containing the crestin gene 
            followed by the mus musculus c Fos minimal promoter and then a 
            rabbit beta globin intron to drive neural crest-specific gene 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4316)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 4316)
  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"; journalName: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: p5E-Crestin-pro (JDW 1320)
FEATURES             Location/Qualifiers
     source          1..4316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          1696..2268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    2362..2485
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2557..2576)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2558..2576)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2581..2597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2581..2597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2699..2718)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2710..3519
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2777..2796)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3666..4254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4155..4174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccga gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgtc tagaggtacc cctcgagccg cagatgttct
      721 agtacccttc ggctaactag ccaatgatta attaaatcag ttaaattaaa aacaatggcc
      781 cttacaggat cttcagcgga aaccatttca cgagactttt aattgactcc tggtcatgtg
      841 ctcgccttgg gcacatcctg ctagaagcac aatttcgtag agacaaagag atctctagaa
      901 actttaatgc atatgaatga aagacaatct cttaaaatgt tagaactaag gcagatgtat
      961 ctttgaataa tatgccataa tgttcctctt atgctgttta ttgcaaatca accaggttaa
     1021 tcaatgtgtt gttttaaccc ttataacaga gtaaaatgtt ctgtatgtgt ctgaaatgta
     1081 tgatgtctgg tattgaacga aagctagtga catttgcaat acgttggtgt aaatgaaaaa
     1141 ctagtagtac aaacagtatt aatcttataa cgtttaatgt aagatagttt caatcttgca
     1201 agagttttac cacttgaaat ctacatgcag tctgttaatt attgaccttc cctacgtcag
     1261 tacggggtgt ggcccgccct tcatggcaac tgctcattgg cagacagtgc catggggacg
     1321 aaccagacca ggtcacttaa aaacaccctg cgacaaagaa actttgcttt ttgcatcagc
     1381 tagcttgcgc atgcttttgc tctctgcccc ctttcgcctg cccggacacc gcgccgccac
     1441 gcgattgggt cactacgaac tttcacgcaa acctcttaag cttcgggact cccgaagctg
     1501 catgaacttc cgcaaaccga ctctcagcag tctgtcacct cagtaaccca ggatatcgtc
     1561 gacccagtga cgtaggaagt ccatccattc acagcgcttc tataaaggcg ccagctgagg
     1621 cgcctactac tccaaccgcg actgcagcga gcaactgagc tcggatccgt ttaaaccgat
     1681 cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta
     1741 aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt
     1801 cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga
     1861 caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc
     1921 ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt
     1981 ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt
     2041 tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc
     2101 tggcgtggaa atattcttat tggtagaaac aactacaccc tggtcatcat cctgcctttc
     2161 tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa
     2221 ccgggcccct ctgctaacca tgttcatgcc ttcttctctt tcctacagct cctgggcaac
     2281 gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tcctcgacac cggtatcgat
     2341 acgcgttacg tagcggccgc tcaagtttgt acaaaaaagt tgaacgagaa acgtaaaatg
     2401 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
     2461 aaacacaaca tatgcagtca ctatgaacca actacttaga tggtattagt gacctgtaga
     2521 attcgagctc tagagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg
     2581 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2641 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2701 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2761 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2821 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2881 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2941 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     3001 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     3061 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     3121 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     3181 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     3241 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     3301 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     3361 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     3421 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     3481 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     3541 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     3601 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3661 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3721 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3781 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3841 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3901 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3961 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4021 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4081 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4141 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4201 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4261 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
