LOCUS       sequence-299711-        5509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXD4L3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5509)
  AUTHORS   Moparthi L, Koch S
  TITLE     A uniform expression library for the exploration of FOX 
            transcription factor biology.
  JOURNAL   Differentiation. 2020 Sep - Oct;115:30-36. doi: 
            10.1016/j.diff.2020.08.002. Epub 2020 Aug 20.
  PUBMED    32858261
REFERENCE   2  (bases 1 to 5509)
  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: 
            "Differentiation. 2020 Sep - Oct"; volume: "115"; pages: "30-36. 
            doi: 10.1016/j.diff.2020.08.002. Epub 2020 Aug 20"
COMMENT     Sequence Label: Flag-FOXD4L3
FEATURES             Location/Qualifiers
     source          1..5509
                     /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"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             94..117
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1561..1695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1585..1604)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1639..1658
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1808..1828)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1808..1826)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1847..1863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1847..1863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1860..1882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1871..1887
                     /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(1895..1901,1902..1919,1920..1925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1940..1961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2078..2095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2249..2837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2329..2348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3008..3799,3800..3868))
                     /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     3631..3650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3869..3973)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3999..4454)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4136..4157)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4348..4367
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4525..5509
                     /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 atcagccacc atggattaca aggacgacga cgataaggga
      121 tcccatcgat tcgaattcaa cttgccaaga gctgagcgcc ttcgctccac accgcagcgc
      181 agcctccggg actccgatgg ggaagacggt aaaatcgatg tcctgggaga ggaggaagat
      241 gaagacgagg tggaagacga ggaggaggag gcgagacagc agttcctaga gcagtcgctc
      301 cagccggggc tgcaggtggc ccggtggggc ggggttgcgc ttccccgaga gcacatcgag
      361 ggcggcggcg gcccgagcga cccctcagag tttggcacca agttcagggc accgccaagg
      421 tctgcggcgg cctctgaaga tgcccggcag ccggcaaagc ccccctactc gtacatcgcg
      481 ctcatcacca tggccatcct gcaaaacccg cacaagcgcc tcacgctcag cggcatctgc
      541 gccttcatta gtggccgctt cccatactac cgccgcaagt tccccgcctg gcagaacagc
      601 atccgccaca acctctcgct gaacgactgc ttcgttaaga tcccccgcga gccgggccgc
      661 ccaggcaagg gcaactactg gagcctggac cccgcctccc aggacatgtt cgacaatggc
      721 agctttctcc ggcgtaggaa gcgtttcaag cgccaccaac tgaccccggg agcccacctg
      781 ccccacccct tccctctacc tgctgcacac gccgccctgc acaaccccca cccaggccct
      841 ctgcttgggg cccctgcccc gccgcagcca gtcccggggg cctaccccaa caccgccccc
      901 gggagacgcc cttacgctct gctgcacccg catcctcttc gctacctact gctctcggcc
      961 cccgtctatg ccggggcacc gaagaaagca gaaggcgcgg acctggcgac cccggcaccc
     1021 ttcccgtgct gcagccctca cttggtcctc agccttggga ggagggcaag ggtctggcgt
     1081 cgccaccggg aggcggatgc atctctttca gcattgagag tattatgcaa ggggtcaggg
     1141 gagcgggtac aggggctgcg cagaatttgt ccccgaccgc gtggagctac tgccacctgc
     1201 tccagcgacc atcaagcctg ttgcatcccc agaccgctgc ccctttgctg caagtgtccg
     1261 ccgccgccgc tgctcggaca attttgcagc aatagcagca gcatcaggag gaggactgcg
     1321 ccaacggctg cgctcccacc aagggcgcgg tgctgggcgg gcacctgtcg gcctcgtcgg
     1381 ccctgctgag gtatcaggca gtggcagagg gctctaggct gacatcgctg gctgcccctt
     1441 tgggcggaga ggggacctca ccagtttttt tagtatcgcc cacgcccagt tccctggcca
     1501 actccgcagg gccctcctag agcctcgagc ctctagaact atagtgagtc gtattacgta
     1561 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     1621 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1681 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1741 gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct
     1801 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     1861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt
     1921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3001 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3061 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3121 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3901 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3961 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     4021 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4081 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4141 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4201 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4261 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4321 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4381 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4441 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4501 gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca
     4561 tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg
     4621 tgccaattca atatggtgga tctggacccc agccaattca atatggcgga cttggcacca
     4681 tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg
     4741 gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg
     4801 gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg caatatccaa
     4861 tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc
     4921 atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt
     4981 cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca
     5041 atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat
     5101 ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga
     5161 cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg
     5221 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
     5281 tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca
     5341 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
     5401 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
     5461 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
