LOCUS       sequence-365714-        6159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible chicken EN2 (mut) activation via mammalian cell 
            transfection or mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  AUTHORS   Rampon C, Gauron C, Lin T, Meda F, Dupont E, Cosson A, Ipendey E, 
            Frerot A, Aujard I, Le Saux T, Bensimon D, Jullien L, Volovitch M, 
            Vriz S, Joliot A
  TITLE     Control of brain patterning by Engrailed paracrine transfer: a new 
            function of the Pbx interaction domain.
  JOURNAL   Development. 2015 May 15;142(10):1840-9. doi: 10.1242/dev.114181. 
            Epub 2015 Apr 29.
  PUBMED    25926358
REFERENCE   2  (bases 1 to 6159)
  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: 
            "10.1242/dev.114181"; journalName: "Development"; date: "2015-05-15-
            15"; volume: "142"; issue: "10"; pages: "1840-9"
COMMENT     Sequence Label: pCSm5En2CER4(#457)
FEATURES             Location/Qualifiers
     source          1..6159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             142..171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             181..210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             220..249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             259..288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1204..1224
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2211..2345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2235..2254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2289..2308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2458..2478)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2458..2476)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2497..2513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2497..2513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2510..2532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2521..2537
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2545..2551,2552..2569,2570..2575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2590..2611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2728..2745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2899..3487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2979..2998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3658..4449,4450..4518))
                     /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     4281..4300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4519..4623)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4649..5104)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4786..4807)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4998..5017
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5175..6159
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggaggagggc ggcaggtcgc cccgggagga ggcggccgaa
      361 ccgcaggagt ccggcgggga cgcggaaccc ggcggagggc gtcgggctct gctgctgccc
      421 cccggcgacc ccccgcaccc ccacccgcac cctcaccgca tcaccaactt cttcatcgac
      481 aacatcctga ggcccgagtt cggtcggagg aaggaagcgg gaggaaccgc cggagaaccc
      541 cgcaggcccg gagcggagag caggaggagc cccgcggccg ccgcgcccgc tcccggagct
      601 ccggtgcccg gcggaggagg cggcggcggg ggtggatcgc cgggccgagg ggagggcggc
      661 cccgccgctc tggccctgca cggagcggcc aagaaggggg gagaccccgc ggcgctggag
      721 gcggccctga aagcgcgggg gctgagcggc gcggagctgt cggtgagctc ggactcggat
      781 agctcccagg ccggctccaa cgccgggaac caacccatgc tttggcccgc ttgggtgtac
      841 tctacaaggt actcggaccg accgtcttca ggtccccgct cccgcaaacc aaagaagaag
      901 aaccccaaca aggaagacaa gcggccccgc accgccttca cggccgagca gctgcagaga
      961 ctcaaggccg agttccagac gaaccgctac ctgacggagc agcgccggca gagcctggcc
     1021 caggagctcg gcctcaacga gtcccagatc aagatctggt tccagaacaa gagagccaag
     1081 atcaagaagg cgacgggcag caagaactcg ctggcggtgc acctcatggc ccaggggctc
     1141 tacaaccact ccaccacggc gaaagacggc aagtcggaca gcgaagcttt ggcgcccggg
     1201 ccagagaacc tgtacttcca ggggctcgag ccctctgccg gagacatgag agctgcaaac
     1261 ctttggccga gcccgctgat gattaaacgc tctaagaaga acagcctggc cttgtccctg
     1321 accgccgacc agatggtcag tgccttgttg gatgccgagc cccccatact ctattccgag
     1381 tatgatccta ccagaccctt cagtgaggct tcgatgatgg gcttactgac caacctggca
     1441 gacagggagc tggttcacat gattaactgg gccaagaggg tgccaggctt tgtggatttg
     1501 accctccacg atcaggtcca cctcctagaa tgtgcctggc tagagatcct gatgattggt
     1561 ctcgtctggc gcagcatgga gcacccagtg aagctactgt ttgctcctaa cttgctcttg
     1621 gacaggaacc agggaaaatg tgtagagggc atggtggaga ttttcgacat gctgctggct
     1681 acatcatctc ggttccgcat gatgaatctg cagggagagg aatttgtgtg cctcaaatct
     1741 attattttgc ttaattctgg agtttacaca tttctgtcca gcaccctgaa gtctctggag
     1801 gaaaaggacc atatccatcg agtgctggac aagatcacag acactttgat tcacctgatg
     1861 gctaaggctg gcctgaccct gcagcagcag catcagagac tcgcacagct tctcctgatc
     1921 ctctcccaca tcaggcacat gagtaacaaa ggcatggagc atctgtactc tatgaagtgc
     1981 aagaacgtgg tgcccctcta tgatctgctg ctggaggcgg cggacgctca ccgcctacat
     2041 gcgcccacta gccgtggagg ggcatccgtg gaggaaacag accaaagcca cttggctact
     2101 gcgggctcta cttcatctca ttccttgcaa aagtattaca tcacggggga ggcagagggt
     2161 ttccctgcca cagcaccagg ttccggcagc gatctgaggg cccgtttgta gatccagaca
     2221 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2281 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2341 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2401 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     2461 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2521 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     2581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4621 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4681 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4741 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4801 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4861 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4921 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4981 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5041 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5101 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5161 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
     5221 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     5281 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     5341 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     5401 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     5461 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     5521 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     5581 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     5641 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     5701 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     5761 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     5821 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     5881 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     5941 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
     6001 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     6061 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     6121 taatgggagg tctatataag caatgctcgt ttagggaac
//
