LOCUS       sequence-296333-        6053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXN1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6053)
  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 6053)
  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-FOXN1
FEATURES             Location/Qualifiers
     source          1..6053
                     /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    2105..2239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2129..2148)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2183..2202
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2352..2372)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2352..2370)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2391..2407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2391..2407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2404..2426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2415..2431
                     /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(2439..2445,2446..2463,2464..2469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2484..2505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2622..2639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2793..3381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2873..2892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3552..4343,4344..4412))
                     /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     4175..4194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4413..4517)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4543..4998)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4680..4701)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4892..4911
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5069..6053
                     /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 tccgtgtcgc tacccccgcc gcagtctgac gtcacgctgc cgggccccac cagactggag
      181 ggcgagcgcc aaggggacct catgcaggca ccgggcctcc caggctcccc tgccccacag
      241 agtaagcatg ccggcttcag ctgctcgtca tttgtgtccg acggccctcc agagaggaca
      301 ccctcactgc ccccacacag cccccgcatt gcgtcaccag ggcccgagca agtccagggc
      361 cactgcccag ccggccccgg ccctgggccc ttcaggctct caccctcaga caagtatcct
      421 ggctttggct ttgaggaggc cgcagcaagc agccctgggc gattcctcaa gggcagccac
      481 gcgcccttcc acccgtacaa gcggcctttc catgaggacg tcttcccaga ggccgagacc
      541 accctggccc tcaaaggaca ctcctttaag accccagggc cgctggaggc cttcgaggag
      601 atcccagtgg acgtggcgga ggccgaggcc ttcctgcctg gcttctcagc agaggcctgg
      661 tgtaacgggc tcccctaccc cagccaggag catggccccc aagtcctggg ttcagaggtc
      721 aaagtcaagc ccccagttct ggagagtggt gctgggatgt tctgctacca gcctcccttg
      781 cagcatatgt actgctcctc ccagcccccc ttccaccagt actcgccagg tggtggcagc
      841 taccccatac cctacctggg ctcctcacac tatcagtacc agcgaatggc accccaggcc
      901 agcaccgatg ggcaccagcc tctcttccca aaacccatct attcctacag catcctcatc
      961 ttcatggccc ttaagaacag taaaactggg agccttcccg tcagcgagat ctacaatttt
     1021 atgacggagc actttcctta cttcaagaca gcacccgatg gctggaagaa ttctgtccgg
     1081 cacaacctat ccctcaacaa gtgcttcgag aaggtggaga acaaatcagg aagttcctcc
     1141 cgcaagggct gcctgtgggc cctcaatccg gccaagatcg acaagatgca agaggagctg
     1201 caaaaatgga agaggaaaga tcccattgct gtgcgcaaaa gcatggccaa gccagaagag
     1261 ctggacagcc tcattggaga caagagagaa aagctgggct ccccactcct gggctgtccg
     1321 ccccctgggc tgtccggctc aggcgccatc cggcccctgg cacccccagc tggcctctcc
     1381 ccaccactgc actcactcca cccagctcca ggccccattc ctggcaagaa ccccctgcag
     1441 gacctactta tggggcacac accctcctgc tatgggcaga catacttgca cctctcacca
     1501 ggcctggccc ctcctggacc cccgcagcca ttgttcccac agccggacgg gcaccttgag
     1561 ctgcgggccc agccaggcac cccccaggac tcgcctctgc ctgcccacac cccacccagc
     1621 cacagtgcca agctactggc cgagccttcc ccagccagga ctatgcacga caccctgctg
     1681 ccagatggag accttggcac tgacctggat gccatcaatc cctcactcac tgacttcgac
     1741 ttccagggaa acctgtggga acagttgaag gatgatagct tggccctcga ccccctggta
     1801 ctggtgacct catccccgac atcatcttcg atgccaccac cccagccacc acctcactgc
     1861 ttcccccctg ggccctgtct gacagagaca ggcagtgggg caggtgactt ggcagccccg
     1921 ggcagtggtg gctccggggc actgggtgac ctgcacctca ccaccctcta ctctgccttt
     1981 atggagctgg agcccacgcc ccccacggcc cctgcaggcc cctctgtgta cctcagcccc
     2041 agctccaagc ccgtggccct ggcatagctc gagcctctag aactatagtg agtcgtatta
     2101 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2161 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2221 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2281 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
     2341 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     2401 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     2461 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2521 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2581 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2641 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2701 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3541 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3601 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3661 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3721 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3781 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3841 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3901 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3961 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4021 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4081 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4141 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4201 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4261 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4321 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4381 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4441 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4501 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4561 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4621 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4681 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4741 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4801 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4861 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4921 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4981 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     5041 cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga
     5101 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
     5161 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
     5221 accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg
     5281 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
     5341 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
     5401 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
     5461 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
     5521 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
     5581 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
     5641 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
     5701 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
     5761 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
     5821 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
     5881 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
     5941 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
     6001 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
