LOCUS       sequence-299559-        5000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXR1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  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 5000)
  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-FOXR1
FEATURES             Location/Qualifiers
     source          1..5000
                     /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    1052..1186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1076..1095)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1130..1149
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1299..1319)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1299..1317)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1338..1354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1338..1354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1351..1373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1362..1378
                     /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(1386..1392,1393..1410,1411..1416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1431..1452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1569..1586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1740..2328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1820..1839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2499..3290,3291..3359))
                     /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     3122..3141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3360..3464)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3490..3945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3627..3648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3839..3858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4016..5000
                     /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 gaacgagctc tttctggcct tcaccacatc tcacctcccc
      181 ttagcggagc agaaacttgc caggtataaa ctccgaattg ttaagccacc aaaattaccc
      241 ctagagaaaa aacccaaccc tgataaggat ggtccagatt atgagcccaa cctctggatg
      301 tgggtaaatc ccaacattgt gtatccccct ggaaagctgg aggtctcagg acgtaggaag
      361 agggaggacc tgacaagcac actcccctcc tctcagccac cccagaagga ggaagatgcc
      421 agctgctcag aggccgcagg ggtggaatca ctgtcccagt cctccagcaa gcggtctccc
      481 cctcggaagc ggtttgcctt ttcccccagc acctgggagc tcacagaaga ggaggaggct
      541 gaggaccagg aagacagctc ctctatggct ctcccatccc ctcacaaaag ggcccccctc
      601 cagagtcgga ggcttcggca agccagcagc caggcgggga ggctctggtc ccggccccct
      661 ctcaattact tccacctaat tgccctggca ttaagaaaca gttccccctg tggcctcaac
      721 gtgcaacaga tctacagttt cactcgaaag cacttcccct ttttccggac ggccccggaa
      781 ggctggaaga atactgtccg tcacaatctc tgttttcgag acagctttga gaaagtgcct
      841 gtcagcatgc agggcggggc cagcacacgg cctcgatctt gcctctggaa gttgaccgag
      901 gagggacacc gccgctttgc ggaggaggcc cgcgccttgg cttccactcg gctagaaagt
      961 atccaacagt gcatgagcca gccagatgtg atgcccttcc tctttgatct ttaactcgag
     1021 cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga
     1081 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1141 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1201 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc
     1261 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     1321 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1381 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2161 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2401 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3361 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3421 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3481 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3541 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3601 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3661 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3721 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3781 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3841 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3901 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     3961 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga
     4021 ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     4081 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     4141 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact
     4201 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     4261 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     4321 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     4381 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     4441 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     4501 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     4561 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     4621 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     4681 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     4741 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     4801 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     4861 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     4921 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     4981 gcaatgctcg tttagggaac
//
