LOCUS       sequence-299702-        5454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXD4L1in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5454)
  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 5454)
  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-FOXD4L1
FEATURES             Location/Qualifiers
     source          1..5454
                     /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    1506..1640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1530..1549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1584..1603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1753..1773)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1753..1771)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1792..1808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1792..1808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1805..1827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1816..1832
                     /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(1840..1846,1847..1864,1865..1870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1885..1906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2023..2040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2194..2782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2274..2293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2953..3744,3745..3813))
                     /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     3576..3595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3814..3918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3944..4399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4081..4102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4293..4312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4470..5454
                     /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 ctcgctccac accgcagcgc
      181 agcctccggg actccgatgg ggaagacggt aaaatcgatg tcctgggaga ggaggaagat
      241 gaagacgagg tggaagacga ggaggaggag gcgagccaga agttcctaga gcagtcgctc
      301 cagccggggc tgcaggtggc ccggtggggc ggggttgcgc ttccccgaga gcacatcgag
      361 ggcggcggcc cgagcgaccc ctcagagttt ggcaccgagt tcagggcacc gccaaggtct
      421 gcggcggcct ctgaagatgc ccggcagccg gcaaagcccc cctactcgta catcgcgctc
      481 atcaccatgg ccatcctgca aagcccgcac aagcgcctca cgctcagcgg catctgcgcc
      541 ttcattagtg gccgcttccc ctactaccgc cgcaagttcc ccgcctggca gaacagcatc
      601 cgccacaacc tctcgctgaa cgactgcttc gtcaagatcc cccgcgagcc gggccaccca
      661 ggcaagggca cctactggag cctggacccc gcctcccagg acatgttcga caatggcagc
      721 tttctccggc gtaggaagcg tttcaagcgc caccaactga ccccgggagc ccacctgccc
      781 caccccttcc ctctacctgc tgcacacgcc gccctgcaca acccccgccc aggccctctg
      841 cttggggccc ctgccctgcc gcagccagtc ccgggggcct accccaacac cgcccccggg
      901 agacgccctt acgctctgct gcacccgcat cctcctcgct acctactgct ctcggccccc
      961 gcctatgccg gggcaccgaa gaaagcagaa ggcgcggacc tggcgacccc cggcaccctt
     1021 cccgtgctgc agccctcact tggtcctcag ccttgggagg agggcaaggg tctggcgtcg
     1081 ccaccgggag gcggatgcat ctctttcagc attgagagta tcatgcaagg ggtcagggga
     1141 gcgggtacag gggctgcgca gagtttgtcc ccgaccgcgt ggagctactg ccccctgctc
     1201 cagcgaccgt caagcctgtc ggacaatttt gcagcaacag cagcagcatc aggaggagga
     1261 ctgcgccaac ggctgcgctc ccaccaaggg cgcggtgctg ggcgggcacc tgtcggccgc
     1321 gtcggcgctg ctgcggtatc aggcggtggc agagggctct aggctgacat cgctggctgc
     1381 ccctttgggc ggagagggga cctcaccagt ttttttagta tcgcccacgc ccagttccct
     1441 ggccaagtcc gcagggccct cctagagcct cgagcctcta gaactatagt gagtcgtatt
     1501 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1561 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1621 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1681 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     1741 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     1801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat
     1861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     3961 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4021 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4081 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4141 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4201 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4261 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4321 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4381 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     4441 tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg
     4501 actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg
     4561 actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg
     4621 caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg
     4681 gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg
     4741 gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata
     4801 tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg tattggccct
     4861 atgccatata gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg
     4921 gcggtcccat ataccatata tggggcttcc taataccgcc catagccact cccccattga
     4981 cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg
     5041 tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc
     5101 attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat
     5161 atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat
     5221 caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat
     5281 tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat
     5341 ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg
     5401 ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
