LOCUS       sequence-360330-        4034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Retinoic Acid activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4034)
  AUTHORS   Rekler D, Kalcheim C
  TITLE     Completion of neural crest cell production and emigration is 
            regulated by retinoic-acid-dependent inhibition of BMP signaling.
  JOURNAL   Elife. 2022 Apr 8;11. pii: 72723. doi: 10.7554/eLife.72723.
  PUBMED    35394423
REFERENCE   2  (bases 1 to 4034)
  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: "Elife."; 
            date: "2022-04-8"; volume: "11. pii: 72723. doi"; pages: " 
            10.7554/eLife.72723"
COMMENT     Sequence Label: pGL3 RARE-RFP
FEATURES             Location/Qualifiers
     source          1..4034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             14..691
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             14..691
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(144..162)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     412..431
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     606..629
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(781..800)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    846..901
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(900..919)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1262..1278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1262..1278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1275..1297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1286..1302
                     /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(1310..1316,1317..1334,1335..1340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1355..1376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1435..1630
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1481..1616
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1636..1770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1673..1692)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1727..1746
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1838..1855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2009..2597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2089..2108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2768..3559,3560..3628))
                     /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     3391..3410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3629..3733)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3838..4034
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3885..4020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aagcttgcta gccatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt
       61 gcgcatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
      121 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
      181 cgcctgggac atcctgtccc ctcagttcca gtacggctcc aaggcctacg tgaagcaccc
      241 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
      301 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
      361 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
      421 gcagaagaag accatgggct gggaggcctc caccgagcgg atgtaccccg aggacggcgc
      481 cctgaagggc gagatcaaga tgaggctgaa gctgaaggac ggcggccact acgacgccga
      541 ggtcaagacc acctacatgg ccaagaagcc cgtgcagctg cccggcgcct acaagaccga
      601 catcaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
      661 cgccgagggc cgccactcca ccggcgccta agaatgcggc cgcactcctc aggtgcaggc
      721 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
      781 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
      841 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
      901 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
      961 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     1021 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     1081 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     1141 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     1201 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     1261 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1321 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1381 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     1441 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     1501 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1561 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     1621 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1741 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     1801 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1861 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1921 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1981 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2041 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2101 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2161 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2221 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2281 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2341 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2401 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2461 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2521 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3721 taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtcgacagtt gggtcatttg
     3781 aaggttagca gcccgggtag ggttcaccga aagttcactc gctcgagatc tgcgatctgc
     3841 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3901 cgcccagttc cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg
     3961 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4021 taggcttttg caaa
//
