LOCUS       sequence-311603-        5509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXD4L6 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-FOXD4L6
FEATURES             Location/Qualifiers
     source          1..5509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(79..98)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(758..1549,1550..1618))
                     /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     1381..1400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1619..1723)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1749..2204)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1886..1907)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2098..2117
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2275..3259
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3294..3312
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3294..3311
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3344..3353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3353..3376
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4820..4954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4844..4863)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4898..4917
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5067..5087)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5067..5085)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5106..5122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5106..5122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5119..5141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5130..5146
                     /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(5154..5160,5161..5178,5179..5184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5199..5220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5337..5354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5508..5509,1..587))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
       61 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
      721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
      781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
      901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1741 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1801 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     1861 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1921 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1981 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2041 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2101 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2161 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     2221 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
     2281 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
     2341 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
     2401 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
     2461 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
     2521 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
     2581 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
     2641 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
     2701 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
     2761 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
     2821 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
     2881 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
     2941 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
     3001 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
     3061 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
     3121 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
     3181 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     3241 caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg
     3301 tgacactata gaatacaagc tacttgttct ttttgcagga tcagccacca tggattacaa
     3361 ggacgacgac gataagggat cccatcgatt cgaattcaac ttgccaagag ctgagcgcct
     3421 tcgctccaca ccgcagcgca gcctccggga ctccgatggg gaagacggta aaatcgatgt
     3481 cctgggagag gaggaagatg aagacgaggt ggaagacgag gaggaggcgg cgagccagca
     3541 gttcctagag cagtcgctcc agccggggct gcaggtggcc cggtggggcg gggttgcgct
     3601 tccccgagag cacatcgagg gcggcggcgg cccgagcgac ccctcagagt ttggcaccaa
     3661 gttcagggca ccgccaaggt ctgcggcggc ctctgaagat gcccggcagc cggcaaagcc
     3721 cccctactcg tacatcgcgc tcatcaccat ggccatcctg caaaacccgc acaagcgcct
     3781 cacgctcagc ggcatctgcg ccttcattag tggccgcttc ccctactacc gccgcaagtt
     3841 ccccgcctgg cagaacagca tccgccacaa cctctcgctg aacgactgct tcgttaagat
     3901 cccccgcgag ccgggccacc caggcaaggg caactactgg agcctggacc ccgcctccca
     3961 agacatgttc gacaatggca gctttctccg gcgtaggaag cgtttcaagc gccaccaact
     4021 gaccccggga gcccacctgc cccacccctt ccctctacct gctgcacacg ccgccctgca
     4081 caacccccac ccaggccctc tgcttggggc ccctgccccg ccgcagccag tcccgggggc
     4141 ctaccccaac accgcccccg ggagacgccc ttacgctctg ctgcacccgc atcctcttcg
     4201 ctacctactg ctctcggccc ccgtctatgc cggggcaccg aagaaagcag aaggcgcggc
     4261 cctggcgacc ccggcaccct tcccgtgctg cagccctcac ttggtcctca gccttgggag
     4321 gagggcaagg gtctggcgtc gccaccggga ggcggatgca tctctttcag cattgagagt
     4381 attatgcaag gggtcagggg agcgggtaca ggggctgcgc agagtttgtc cccgaccgcg
     4441 tggagctact gccacctgct ccagcgacca tcaagcctgt tgcatcccca gaccgctgcc
     4501 cctttgctgc aagtgtccgc cgccgccgct gctcggacaa ttttgcagca atagcagcag
     4561 catcaggagg aggactgcgc caacggctgc gctcccacca agggcgcggt gctgggcggg
     4621 cacctgtcgg cctcgtcggc cctgctgagg tatcaggcag tggcagaggg ctctaggctg
     4681 acatcgctgg ctgccccttt gggcggagag gggacctcac cagttttttt agtatcgccc
     4741 acgcccagtt ccctggccaa ctccgcaggg ccctcctaga gcctcgagcc tctagaacta
     4801 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     4861 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4921 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4981 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     5041 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     5101 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5161 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5221 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5281 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5341 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5401 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5461 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgtttt
//
