LOCUS       sequence-421917-        6651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6651)
  AUTHORS   Seoane J, Le HV, Shen L, Anderson SA, Massague J
  TITLE     Integration of Smad and forkhead pathways in the control of 
            neuroepithelial and glioblastoma cell proliferation.
  JOURNAL   Cell. 2004 Apr 16. 117(2):211-23.
  PUBMED    15084259
REFERENCE   2  (bases 1 to 6651)
  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: "Cell. 2004 
            Apr 16. 117(2):211-23."
COMMENT     Sequence Label: Flag FoxO3
FEATURES             Location/Qualifiers
     source          1..6651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(953..1744,1745..1813))
                     /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     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1945..2400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2032..2051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2242..2263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2526..2548
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2540..2557
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2541..2557
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2718..3097
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3098..3301
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3252..3272
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3336..3359
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5413..6035
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5612..5631)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6059..6079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6224..6358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6410..6429)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6411..6429)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6443..6459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6443..6459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6456..6478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6467..6483
                     /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(6491..6497,6498..6515,6516..6521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6536..6557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2101 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2161 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2281 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2341 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2401 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2461 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     2521 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
     2581 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     2641 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     2701 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     2761 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2821 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2881 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2941 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
     3001 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
     3061 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
     3121 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     3181 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
     3241 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3301 tcgtttagtg aaccgtcaga attgcagtga ccatggacta caaggacgac gatgacaagg
     3361 aattcagatc tggtaccatg gcagaggcac cggcttcccc ggccccgctc tctccgctcg
     3421 aagtggagct ggacccggag ttcgagcccc agagccgtcc gcgatcctgt acgtggcccc
     3481 tgcaaaggcc ggagctccaa gcgagccctg ccaagccctc gggggagacg gccgctgact
     3541 ccatgatccc cgaggaggag gacgatgaag acgacgagga cggcggggga cgggccggct
     3601 cggccatggc gatcggcggc ggcggcggga gcggcacgct gggctccggg ctgctccttg
     3661 aggactcggc ccgggtgctg gcacccggag ggcaagaccc cgggtctggg ccagccaccg
     3721 cggcgggcgg gctgagcggg ggtacacagg cgctgctgca gcctcagcaa ccgctgccac
     3781 cgccgcagcc gggggcggct gggggctccg ggcagccgag gaaatgttcg tcgcggcgga
     3841 acgcctgggg aaacctgtcc tacgcggacc tgatcacccg cgccatcgag agctccccgg
     3901 acaaacggct cactctgtcc cagatctacg agtggatggt gcgttgcgtg ccctacttca
     3961 aggataaggg cgacagcaac agctctgccg gctggaagaa ctccatccgg cacaacctgt
     4021 cactgcatag tcgattcatg cgggtccaga atgagggaac tggcaagagc tcttggtgga
     4081 tcatcaaccc tgatgggggg aagagcggaa aagccccccg gcggcgggct gtctccatgg
     4141 acaatagcaa caagtatacc aagagccgtg gccgcgcagc caagaagaag gcagccctgc
     4201 agacagcccc cgaatcagct gacgacagtc cctcccagct ctccaagtgg cctggcagcc
     4261 ccacgtcacg cagcagtgat gagctggatg cgtggacgga cttccgttca cgcaccaatt
     4321 ctaacgccag cacagtcagt ggccgcctgt cgcccatcat ggcaagcaca gagttggatg
     4381 aagtccagga cgatgatgcg cctctctcgc ccatgctcta cagcagctca gccagcctgt
     4441 caccttcagt aagcaagccg tgcacggtgg aactgccacg gctgactgat atggcaggca
     4501 ccatgaatct gaatgatggg ctgactgaaa acctcatgga cgacctgctg gataacatca
     4561 cgctcccgcc atcccagcca tcgcccactg ggggactcat gcagcggagc tctagcttcc
     4621 cgtataccac caagggctcg ggcctgggct ccccaaccag ctcctttaac agcacggtgt
     4681 tcggaccttc atctctgaac tccctacgcc agtctcccat gcagaccatc caagagaaca
     4741 agccagctac cttctcttcc atgtcacact atggtaacca gacactccag gacctgctca
     4801 cttcggactc acttagccac agcgatgtca tgatgacaca gtcggacccc ttgatgtctc
     4861 aggccagcac cgctgtgtct gcccagaatt cccgccggaa cgtgatgctt cgcaatgatc
     4921 cgatgatgtc ctttgctgcc cagcctaacc agggaagttt ggtcaatcag aacttgctcc
     4981 accaccagca ccaaacccag ggcgctcttg gtggcagccg tgccttgtcg aattctgtca
     5041 gcaacatggg cttgagtgag tccagcagcc ttgggtcagc caaacaccag cagcagtctc
     5101 ctgtcagcca gtctatgcaa accctctcgg actctctctc aggctcctcc ttgtactcaa
     5161 ctagtgcaaa cctgcccgtc atgggccatg agaagttccc cagcgacttg gacctggaca
     5221 tgttcaatgg gagcttggaa tgtgacatgg agtccattat ccgtagtgaa ctcatggatg
     5281 ctgatgggtt ggattttaac tttgattccc tcatctccac acagaatgtt gttggtttga
     5341 acgtggggaa cttcactggt gctaagcagg cctcatctca gagctgggtg ccaggctgat
     5401 ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     5461 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     5521 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     5581 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     5641 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     5701 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     5761 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     5821 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     5881 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg
     5941 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca
     6001 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg
     6061 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6241 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6301 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6361 ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga
     6421 gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6481 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6541 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6601 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg c
//
