LOCUS       sequence-299545-        6074 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of Flag tagged human FOXO1A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  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 6074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-FOXO1A
FEATURES             Location/Qualifiers
     source          1..6074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1597..1731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1621..1640)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1675..1694
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1844..1864)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1844..1862)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1883..1899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1883..1899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1907..1923
                     /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(1931..1937,1938..1955,1956..1961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1976..1997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2114..2131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2285..2873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2365..2384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3044..3835,3836..3904))
                     /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     3667..3686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3905..4009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4035..4490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4384..4403
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4561..5545
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        5580..5598
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5580..5597
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      5630..5639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5639..5662
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tgccccccgc cgccgctggg ccgctcgcgg ggcagccgcg caagagcagc tcgtcccgcc
       61 gcaacgcgtg gggcaacctg tcctacgccg acctcatcac caaggccatc gagagctcgg
      121 cggagaagcg gctcacgctg tcgcagatct acgagtggat ggtcaagagc gtgccctact
      181 tcaaggataa gggtgacagc aacagctcgg cgggctggaa gaattcaatt cgtcataatc
      241 tgtccctaca cagcaagttc attcgtgtgc agaatgaagg aactggaaaa agttcttggt
      301 ggatgctcaa tccagagggt ggcaagagcg ggaaatctcc taggagaaga gctgcatcca
      361 tggacaacaa cagtaaattt gctaagagcc gaagccgagc tgccaagaag aaagcatctc
      421 tccagtctgg ccaggagggt gctggggaca gccctggatc acagttttcc aaatggcctg
      481 caagccctgg ctctcacagc aatgatgact ttgataactg gagtacattt cgccctcgaa
      541 ctagctcaaa tgctagtact attagtggga gactctcacc cattatgacc gaacaggatg
      601 atcttggaga aggggatgtg cattctatgg tgtacccgcc atctgccgca aagatggcct
      661 ctactttacc cagtctgtct gagataagca atcccgaaaa catggaaaat cttttggata
      721 atctcaacct tctctcatca ccaacatcat taactgtttc gacccagtcc tcacctggca
      781 ccatgatgca gcagacgccg tgctactcgt ttgcgccacc aaacaccagt ttgaattcac
      841 ccagcccaaa ctaccaaaaa tatacatatg gccaatccag catgagccct ttgccccaga
      901 tgcctataca aacacttcag gacaataagt cgagttatgg aggtatgagt cagtataact
      961 gtgcgcctgg actcttgaag gagttgctga cttctgactc tcctccccat aatgacatta
     1021 tgacaccagt tgatcctggg gtagcccagc ccaacagccg ggttctgggc cagaacgtca
     1081 tgatgggccc taattcggtc atgtcaacct atggcagcca ggcatctcat aacaaaatga
     1141 tgaatcccag ctcccatacc caccctggac atgctcagca gacatctgca gttaacgggc
     1201 gtcccctgcc ccacacggta agcaccatgc cccacacctc gggtatgaac cgcctgaccc
     1261 aagtgaagac acctgtacaa gtgcctctgc cccaccccat gcagatgagt gccctggggg
     1321 gctactcctc cgtgagcagc tgcaatggct atggcagaat gggccttctc caccaggaga
     1381 agctcccaag tgacttggat ggcatgttca ttgagcgctt agactgtgac atggaatcca
     1441 tcattcggaa tgacctcatg gatggagata cattggattt taactttgac aatgtgttgc
     1501 ccaaccaaag cttcccacac agtgtcaaga caacgacaca tagctgggtg tcaggctagc
     1561 tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
     1621 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1681 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1741 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
     1801 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     1861 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1921 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
     1981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4021 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4081 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4141 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4201 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4261 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4321 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4381 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4441 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     4501 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4561 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
     4621 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
     4681 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
     4741 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
     4801 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
     4861 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
     4921 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
     4981 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
     5041 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
     5101 tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat
     5161 tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg
     5221 acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca
     5281 atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt
     5341 acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg
     5401 gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat
     5461 gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta
     5521 tataagcaat gctcgtttag ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga
     5581 tttaggtgac actatagaat acaagctact tgttcttttt gcaggatcag ccaccatgga
     5641 ttacaaggac gacgacgata agggatccgc cgaggcgcct caggtggtgg agatcgaccc
     5701 ggacttcgag ccgctgcccc ggccgcgctc gtgcacctgg ccgctgccca ggccggagtt
     5761 tagccagtcc aactcggcca cctccagccc ggcgccgtcg ggcagcgcgg ctgccaaccc
     5821 cgacgccgcg gcgggcctgc cctcggcctc ggctgccgct gtcagcgccg acttcatgag
     5881 caacctgagc ttgctggagg agagcgagga cttcccgcag gcgcccggct ccgtggcggc
     5941 ggcggtggcg gcggcggccg ccgcggccgc caccgggggg ctgtgcgggg acttccaggg
     6001 cccggaggcg ggctgcctgc acccagcgcc accgcagccc ccgccgcccg ggccgctgtc
     6061 gcagcacccg ccgg
//
