LOCUS       sequence-299347-        5219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXH1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5219)
  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 5219)
  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-FOXH1
FEATURES             Location/Qualifiers
     source          1..5219
                     /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    1271..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1295..1314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1349..1368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1518..1538)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1518..1536)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1557..1573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1557..1573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1570..1592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1581..1597
                     /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(1605..1611,1612..1629,1630..1635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1650..1671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1788..1805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1959..2547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2039..2058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2718..3509,3510..3578))
                     /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     3341..3360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3579..3683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3709..4164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3846..3867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4058..4077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4235..5219
                     /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 tcgaattcgg gccctgcagc ggctcccgcc tggggccccc agaggcagag
      181 tcgccctccc agccccctaa gaggaggaag aagaggtacc tgcgacatga caagcccccc
      241 tacacctact tggccatgat cgccttggtg attcaggccg ctccctcccg cagactgaag
      301 ctggcccaga tcatccgtca ggtccaggcc gtgttcccct tcttcaggga agactacgag
      361 ggctggaaag actccattcg ccacaacctt tcctccaacc gatgcttccg caaggtgccc
      421 aaggaccctg caaagcccca ggccaagggc aacttctggg cggtcgacgt gagcctgatc
      481 ccagctgagg cgctccggct gcagaacacc gccctgtgcc ggcgctggca gaacggaggt
      541 gcgcgtggag ccttcgccaa ggacctgggc ccctacgtgc tgcacggccg gccataccgg
      601 ccgcccagtc ccccgccacc acccagtgag ggcttcagca tcaagtccct gctaggaggg
      661 tccggggagg gggcaccctg gccggggcta gctccacaga gcagcccagt tcctgcaggc
      721 acagggaaca gtggggagga ggcggtgccc accccacccc ttccctcttc tgagaggcct
      781 ctgtggcccc tctgccccct tcctggcccc acgagagtgg agggggagac tgtgcagggg
      841 ggagccatcg ggccctcaac cctctcccca gagcctaggg cctggcctct ccacttactg
      901 cagggcaccg cagttcctgg gggacggtcc agcgggggac acagggcctc cctctggggg
      961 cagctgccca cctcctactt gcctatctac actcccaatg tggtaatgcc cttggcacca
     1021 ccacccacct cctgtcccca gtgtccgtca accagccctg cctactgggg ggtggcccct
     1081 gaaacccgag ggcccccagg gctgctctgc gatctagacg ccctcttcca aggggtgcca
     1141 cccaacaaaa gcatctacga cgtttgggtc agccaccctc gggacctggc ggcccctggc
     1201 ccaggctggc tgctctcctg gtgcagcctg tgactcgagc ctctagaact atagtgagtc
     1261 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1321 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1381 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1441 tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg
     1501 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     1561 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg
     1621 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1681 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1741 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1801 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1861 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3301 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3661 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     3721 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3781 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3841 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3901 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3961 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4021 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4081 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4141 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     4201 ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
     4261 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
     4321 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
     4381 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
     4441 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
     4501 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
     4561 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
     4621 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
     4681 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
     4741 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
     4801 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
     4861 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
     4921 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
     4981 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
     5041 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
     5101 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
     5161 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
