LOCUS       sequence-299560-        5061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXR2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5061)
  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 5061)
  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-FOXR2
FEATURES             Location/Qualifiers
     source          1..5061
                     /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    1113..1247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1137..1156)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1191..1210
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1360..1380)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1360..1378)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1399..1415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1399..1415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1412..1434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1423..1439
                     /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(1447..1453,1454..1471,1472..1477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1492..1513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1630..1647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1801..2389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1881..1900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2560..3351,3352..3420))
                     /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     3183..3202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3421..3525)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3551..4006)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3688..3709)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3900..3919
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4077..5061
                     /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 tcgaattcga cttaaaacta aaagactgtg aattttggta tagtctccat
      181 ggccaggtcc cagggctgct ggactgggac atgaggaatg agttatttct gccttgtacc
      241 acagaccagt gctctttagc tgagcaaatc cttgccaaat acagagtcgg agtaatgaag
      301 cccccagaaa tgcctcagaa gaggagaccc agtcctgatg gagatggtcc tccctgtgaa
      361 cccaatctgt ggatgtgggt ggaccccaat atcctgtgcc cccttggcag ccaggaggcc
      421 ccaaagccca gtggaaaaga ggatctgaca aacatttctc ctttccctca gcccccacaa
      481 aaagacgaag ggtctaactg ctcagaggac aaagtggtag agtctctgcc atcttcctcc
      541 agtgagcagt ctcctttaca gaagcagggt atccattccc ccagtgactt tgagctcaca
      601 gaagaggagg ctgaggaacc agacgacaac tccctccagt cccctgaaat gaaatgttac
      661 cagagccaga aactatggca aatcaacaac caagagaagt cctggcaaag gccccctctc
      721 aattgtagcc accttattgc cctagcatta agaaacaacc cccactgtgg cctcagtgtg
      781 caggagatct acaatttcac ccgacagcat ttcccctttt tctggacagc tccggatggc
      841 tggaagagca ccattcatta caacctctgc ttcctggaca gctttgagaa ggtgccagac
      901 agccttaagg atgaagataa tgcaagacct cgctcttgcc tttggaagct cactaaggag
      961 gggcaccgcc gcttttggga ggagactcgt gccttagcct ttgctcaaag ggagagaatc
     1021 caagagtgca tgagtcagcc agagttgttg acctctctct ttgatctttg aaatgctcga
     1081 gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
     1141 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1201 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1261 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
     1321 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     1381 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1441 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3541 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3661 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3721 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3781 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3841 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3901 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3961 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     4021 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
     4081 accgccaatt caatatggcg tatatggact catgccaatt caatatggtg gatctggacc
     4141 tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc
     4201 ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac
     4261 tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc
     4321 ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta
     4381 tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg
     4441 ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct
     4501 attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa
     4561 taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt
     4621 gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg
     4681 gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt
     4741 caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga
     4801 cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat
     4861 tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa
     4921 atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg
     4981 caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata
     5041 agcaatgctc gtttagggaa c
//
