LOCUS       sequence-299551-        5592 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of Flag tagged human FOXO6 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5592)
  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 5592)
  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-FOXO6
FEATURES             Location/Qualifiers
     source          1..5592
                     /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    1644..1778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1668..1687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1722..1741
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1891..1911)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1891..1909)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1930..1946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1943..1965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1954..1970
                     /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(1978..1984,1985..2002,2003..2008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2023..2044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2161..2178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2332..2920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2412..2431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3091..3882,3883..3951))
                     /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     3714..3733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3952..4056)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4082..4537)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4219..4240)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4431..4450
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4608..5592
                     /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 tcgaattcgc tgcgaagctg cgagcccatc aggtggacgt ggaccctgac
      181 ttcgctcctc agagccggcc tagatcctgt acatggcccc tgcctcagcc tgacttggcc
      241 ggcgacgaag acggagctct gggagcaggg gttgctgaag gcgctgaaga ctgcggacct
      301 gagagaaggg ctacagcccc tgctatggcc ccagctcctc ccctgggagc tgaggttgga
      361 cctctgcgga aagccaagag ctctaggcgg aacgcctggg ggaacctgtc ctacgccgac
      421 ctgatcacca aagccatcga gagcgccccc gacaagcggc tgaccctgtc ccagatctac
      481 gactggatgg tgcggtacgt gccctacttc aaggataaag gcgacagcaa cagctccgcc
      541 ggctggaaga actccatccg gcacaacctg tccctgcaca cccggttcat ccgcgtgcag
      601 aacgagggca ccggcaagag ttcctggtgg atgctgaacc ccgagggcgg aaagacaggg
      661 aagacccctc ggagaagggc cgtgtccatg gacaacgggg ccaagttcct gcgcatcaag
      721 ggcaaggcca gcaagaagaa gcagctgcaa gctcccgaga gaagccctga tgacagctcc
      781 cctagtgctc ctgctcctgg gcctgtgcct gctgcagcta agtgggccgc tagccctgcc
      841 tctcacgcta gcgatgacta tgaggcttgg gctgatttcc gcggcggcgg aagacccctg
      901 ctgggggaag ctgctgaact ggaggacgac gaagccctgg aggccctggc tccatcatct
      961 cctctgatgt acccaagccc tgccagcgct ctgtctcctg ctctgggctc tcgctgtcct
     1021 ggtgaactgc ccagacttgc cgaactggga ggacctcttg gactgcacgg aggcggcgga
     1081 gctggactgc ctgaaggact gttagacgga gcccaggacg cttacggacc tagggccgct
     1141 cctaggcctg gacctgttct tggtgcccct ggggagctgg ctcttgctgg agcagctgct
     1201 gcttatcctg gcaaaggagc cgccccttac gctcctcccg ctccctctcg cagtgcctta
     1261 gctcacccca tcagccttat gaccctgcct ggcgaggctg gagctgctgg acttgcacct
     1321 cctggacatg ctgctgcttt cggaggccct cctggcggac tgctgctgga tgctcttcct
     1381 gggccttatg ctgctgctgc cgctggacct ttaggagctg cccccgacag attccccgct
     1441 gacctggacc tggacatgtt cagcgggagc ctggagtgcg acgtggagtc catcatcctg
     1501 aacgacttca tggacagcga cgaaatggac ttcaacttcg attctgccct gcctcctcct
     1561 cctcccggcc tggccggggc ccctccccct aaccagagtt gggtgccggg ctgatctaga
     1621 actatagtga gtcgtattac gtagatccag acatgataag atacattgat gagtttggac
     1681 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1741 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1801 ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa
     1861 tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     1921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1981 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4081 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4141 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4201 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4261 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4321 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4381 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4441 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4501 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     4561 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat
     4621 tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat
     4681 tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat
     4741 tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac
     4801 tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa
     4861 gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg
     4921 atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca
     4981 atactatgta ttggccctat gccatatagt attccatata tgggttttcc tattgacgta
     5041 gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca
     5101 tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata
     5161 tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc
     5221 ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat
     5281 ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg
     5341 gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg
     5401 aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc
     5461 gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc
     5521 gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct
     5581 cgtttaggga ac
//
