LOCUS       sequence-299544-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXN3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  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 5571)
  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-FOXN3
FEATURES             Location/Qualifiers
     source          1..5571
                     /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    1623..1757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1647..1666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1701..1720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1870..1890)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1870..1888)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1909..1925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1909..1925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1922..1944)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1933..1949
                     /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(1957..1963,1964..1981,1982..1987))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2002..2023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2140..2157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2311..2899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2391..2410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3070..3861,3862..3930))
                     /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     3693..3712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3931..4035)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4061..4516)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4198..4219)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4410..4429
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4587..5571
                     /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 tccagtcatg cctcccagta agaagccaga aagctcagga
      181 attagtgtct ccagtggact gagtcagtgt tacgggggca gcggtttctc caaggccctt
      241 caggaagacg atgacctcga cttttctctg cctgacatcc gattagaaga gggggccatg
      301 gaagatgaag agctgaccaa cctgaactgg ctgcacgaga gcaagaactt gctgaagagc
      361 tttggggagt cggtcctcag gagtgtcagc cccgtccagg acctggacga tgacaccccc
      421 ccatcccctg cccactctga catgccctac gatgccaggc agaaccccaa ctgcaaaccc
      481 ccctactcct tcagctgcct catatttatg gccatcgagg actctccaac caagcgcctg
      541 ccagtgaagg atatctacaa ctggatcttg gaacattttc cgtattttgc aaatgcacct
      601 actgggtgga aaaactcagt gagacacaat ttatcattga ataagtgttt taagaaagtg
      661 gacaaagaga ggagtcagag tattgggaaa gggtcgttgt ggtgcataga cccagagtat
      721 agacaaaatc taattcaggc tttgaaaaag acaccttatc acccacaccc acacgtgttc
      781 aatacacctc ccacctgtcc tcaggcatat caaagcacat caggtccacc catctggccg
      841 ggcagtacct tcttcaagag aaatggagcc cttctccaag ttcctccagg agtgatccaa
      901 aatggagcgc gggtcctgag ccgagggctg tttcctggcg tgcggccgct gccaatcact
      961 cccattgggg tgacagcggc catgaggaat ggcatcacca gctgccggat gcggactgag
     1021 agtgagccat cttgtggctc cccagtggtc agcggagacc ccaaggagga tcacaactac
     1081 agcagtgcca agtcctccaa cgcccggagc acctcgccca ccagcgactc catctcctcc
     1141 tcctcctcct cagccgacga ccactatgag tttgccacca aggggagcca ggagggcagc
     1201 gagggcagcg aggggagctt ccggagccac gagagcccca gcgacacgga agaggacgac
     1261 aggaagcaca gccagaagga gcccaaggat tctctggggg acagcgggta cgcatcccag
     1321 cacaagaagc gccagcactt cgccaaggcc aggaaggtcc ccagcgacac actgcccctc
     1381 aaaaagagac gcaccgaaaa gccccccgag agcgatgatg aggagatgaa agaagcggca
     1441 gggtccctcc tgcacttagc agggatccgg tcctgtttga ataacatcac caatcggacg
     1501 gcaaaggggc agaaagagca aaaggaaacc acaaaaaatt aaaaacaagt cactgatttg
     1561 ttttgaactt acgaccattt ggtttctcga gcctctagaa ctatagtgag tcgtattacg
     1621 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1681 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1741 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1801 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     1861 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     1921 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     1981 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2041 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2101 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2161 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2221 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2281 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2341 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2401 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2461 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2521 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2581 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2641 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2701 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2761 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2821 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2881 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2941 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3001 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3061 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3121 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3181 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3241 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3301 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3361 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3421 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3481 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3541 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3601 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3661 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3721 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3781 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3841 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3901 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3961 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4021 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     4081 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4141 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4201 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4261 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4321 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4381 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4441 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4501 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4561 gtgcgggcct cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact
     4621 catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact
     4681 cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac
     4741 catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca
     4801 cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca
     4861 tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc
     4921 aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg
     4981 ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg
     5041 gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt
     5101 caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat
     5161 atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt
     5221 gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc
     5281 cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa
     5341 tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg
     5401 cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc
     5461 cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc
     5521 ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
