LOCUS       sequence-299543-        5426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXN2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5426)
  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 5426)
  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-FOXN2
FEATURES             Location/Qualifiers
     source          1..5426
                     /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    1478..1612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1502..1521)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1556..1575
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1725..1745)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1725..1743)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1764..1780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1764..1780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1777..1799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1788..1804
                     /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(1812..1818,1819..1836,1837..1842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1857..1878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1995..2012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2166..2754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2246..2265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2925..3716,3717..3785))
                     /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     3548..3567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3786..3890)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3916..4371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4053..4074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4265..4284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4442..5426
                     /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 tccggtccag taattggaat gactccagat aagagagctg aaaccccagg agctgaaaag
      181 attgcaggat taagccagat ttacaaaatg ggaagcttgc ctgaagctgt tgatgctgcc
      241 aggccgaagg ccactctagt ggacagtgag tcagcagatg atgaactcac aaacttgaac
      301 tggcttcatg aaagcactaa tcttctaaca aacttcagcc tcggaagtga gggtcttcca
      361 attgttagtc cattgtatga catagaggga gatgatgtgc catcctttgg accagcttgc
      421 taccagaacc cagaaaaaaa atcagcgact tcaaagcccc catactcctt tagtcttctc
      481 atttatatgg ccattgagca ctctccaaat aaatgtttgc ctgtcaaaga aatttatagc
      541 tggattctgg accattttcc atattttgct actgcaccaa caggctggaa gaattctgtt
      601 cgacataatc tgtccctgaa taaatgtttt cagaaagtgg aaagaagcca tggcaaggtt
      661 aatggaaaag gttccttatg gtgtgttgat ccggaatata aacccaatct tatccaggca
      721 ctgaagaagc aacctttttc ttcagcatct tcacaaaatg gttctttatc acctcactat
      781 ttaagctctg taatcaagca gaaccaggtg cgaaacctca aagaatctga tattgatgct
      841 gctgctgcaa tgatgctttt aaatacttct atagaacaag gaattttaga atgtgagaag
      901 cctcttcctc ttaaaacagc attgcaaaaa aagaggagtt acggcaatgc atttcatcat
      961 cccagtgctg tacgattaca agagagtgat tctttagcca ccagcattga tccaaaagaa
     1021 gatcacaatt acagtgcaag tagcatggca gcacagcgtt gtgcatccag gtctagcgtg
     1081 tcttctctgt cttctgtgga tgaggtatat gaatttatcc caaagaatag tcacgtggga
     1141 agtgatggca gtgaaggatt tcacagtgaa gaagatacag acgttgatta tgaagatgat
     1201 cctcttggag acagtggcta tgcatcacag ccttgtgcaa aaatctctga aaaagggcag
     1261 tcaggcaaaa agatgcgaaa acagacatgt caagaaattg atgaggagct caaagaggca
     1321 gctggatctc tgctccacct tgctggaatt cgtacatgtt taggttccct aataagtact
     1381 gcaaagacac aaaatcaaaa gcaacggaaa aaatagaaat acttaaagtg tggcaatact
     1441 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     1501 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1561 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1621 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     1681 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     1741 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1801 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     1861 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1921 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1981 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2041 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2101 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2161 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2581 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3901 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3961 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4021 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4081 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4141 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4201 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4261 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4321 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4381 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4441 agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct
     4501 ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct
     4561 ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct
     4621 ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct
     4681 tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt
     4741 ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct
     4801 attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt
     4861 ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt
     4921 cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc
     4981 ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa
     5041 ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt
     5101 gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc
     5161 aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag
     5221 tacattacta ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc
     5281 ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa
     5341 tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct
     5401 atataagcaa tgctcgttta gggaac
//
