LOCUS       sequence-174342-        6791 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 6791)
  AUTHORS   Kitamura YI, Kitamura T, Kruse JP, Raum JC, Stein R, Gu W, Accili D
  TITLE     FoxO1 protects against pancreatic beta cell failure through NeuroD 
            and MafA induction.
  JOURNAL   Cell Metab. 2005 Sep . 2(3):153-63.
  PUBMED    16154098
REFERENCE   2  (bases 1 to 6791)
  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 Metab.
            2005 Sep . 2(3):153-63."
COMMENT     Sequence Label: FLAG-Foxo1(pCMV5)
FEATURES             Location/Qualifiers
     source          1..6791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     141..158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        319..698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        699..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     853..873
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..960
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             988..1011
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3131..3753
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3330..3349)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3777..3797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3782..4111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3962..4097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4149..4168)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4150..4168)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4182..4198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4182..4198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4195..4217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4206..4222
                     /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(4230..4236,4237..4254,4255..4260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4275..4296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4413..4430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4584..5172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4664..4683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5343..6134,6135..6203))
                     /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     5966..5985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6204..6308)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6335..6763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6422..6441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6632..6653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga
      181 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
      241 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
      301 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      541 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
      601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
      661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      721 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
      841 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      901 ctcgtttagt gaaccgtcag aattgccatg gagcagaagc tgatcagcga ggaggacctg
      961 aacgaattca gatctggtac cgccatggac tacaaggacg acgatgacaa ggccgaggcg
     1021 ccccaggtgg tggagaccga cccggacttc gagccgctgc cccggcagcg ctcctgtacc
     1081 tggccgctgc ccaggccgga gtttaaccag tccaactcga ccacctccag tccggcgccg
     1141 tcgggcggcg cggccgccaa ccccgacgcc gcggcgagcc tggcctcggc gtccgctgtc
     1201 agcaccgact ttatgagcaa cctgagcctg ctggaggaga gtgaggactt cgcgcgggcg
     1261 ccaggctgcg tggccgtggc ggcggcggct gcggccagca ggggcctgtg cggggacttc
     1321 cagggccccg aggcgggctg cgtgcaccca gcgccgccac agcccccacc gaccgggccg
     1381 ctgtcgcagc ccccacccgt gcctccctcc gctgccgccg ccgcggggcc actcgcggga
     1441 cagccgcgca agaccagctc gtcgcgccgc aacgcgtggg gcaacctgtc gtacgccgac
     1501 ctcatcacca aggccatcga gagctcagcc gagaagaggc tcaccctgtc gcagatctac
     1561 gagtggatgg tgaagagcgt gccctacttc aaggataagg gcgacagcaa cagctcggcg
     1621 ggctggaaga attcaattcg ccacaatctg tcccttcaca gcaggtttat tcgagtgcag
     1681 aatgaaggaa ctggaaagag ttcttggtgg atgctcaatc cagagggagg caagagcgga
     1741 aaatcacccc ggagaagagc tgcgtccatg gacaacaaca gtaaatttgc taagagccga
     1801 gggcgggctg ctaagaaaaa agcatctctc cagtctgggc aagagggtcc tggagacagc
     1861 cctgggtctc agttttctaa gtggcctgcg agtcctgggt cccacagcaa cgatgacttt
     1921 gataactgga gtacatttcg tcctcgaacc agctcaaatg ctagtaccat cagtgggaga
     1981 ctttctccca tcatgacaga gcaggatgac ctgggagatg gggacgtgca ttccctggtg
     2041 tatccaccct ctgctgccaa gatggcgtct acgctgccca gtctgtctga aatcagcaat
     2101 ccagaaaaca tggagaacct tctggataat ctcaaccttc tctcgtcccc aacatcttta
     2161 actgtgtcca cccagtcctc gcctggcagc atgatgcagc agacaccatg ctattcgttt
     2221 gcaccgccaa acaccagtct aaattcaccc agtccaaact actcaaagta cacatacggc
     2281 caatccagca tgagcccttt gccccagatg cctatgcaga cacttcagga cagcaaatca
     2341 agttacggag gattgaacca gtataactgt gccccaggac tcttgaaaga gttgttgact
     2401 tctgactctc ctccccacaa tgacattatg tcaccggttg atcccggagt ggcccaaccc
     2461 aacagtcggg tcctgggcca aaatgtaatg atgggcccta attcggtcat gccagcgtat
     2521 ggcagccagg catctcataa caaaatgatg aaccccagct cccacaccca ccctggacat
     2581 gcacagcaaa cggcttcggt caacggccgt accctgcccc atgtggtgaa caccatgcct
     2641 cacacatctg ccatgaaccg cttgaccccc gtgaagacac ctttacaagt gcctctgtcc
     2701 caccccatgc agatgagtgc cctgggcagc tactcctcgg tgagcagctg caatggctat
     2761 ggtaggatgg gtgtcctcca ccaggagaag ctcccaagtg acttggatgg catgtttatt
     2821 gagcgcttgg actgtgacat ggagtccatc attcggaatg accccatgga tggagatacc
     2881 ttggatttta actttgataa tgtgttgccc aaccaaagct tcccacacag tgtcaagact
     2941 acaacacaca gctgggtgtc aggctaagag ttagtgagca ggctacattt aaaagtcctt
     3001 cagattgtct gacagcagga actgaggagc agtccaaaga tgcccttcac ccctccttat
     3061 agttttcaag attaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaagcggcc gctgaattct
     3121 agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     3181 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     3241 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     3301 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     3361 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     3421 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     3481 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3541 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3601 tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct
     3661 acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg
     3721 cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa
     3781 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3841 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3901 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3961 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4021 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4081 aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa
     4141 agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg
     4201 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4261 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4321 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4381 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4441 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4501 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4561 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4621 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4681 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4741 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4801 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4861 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4921 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4981 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5041 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5101 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5161 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5221 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5281 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5341 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5401 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5461 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5521 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5581 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5641 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5701 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5761 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5821 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5881 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5941 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6001 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6061 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6121 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6181 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6241 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6301 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     6361 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     6421 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6481 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6541 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6601 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6661 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     6721 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     6781 cttacaattt c
//
