LOCUS       sequence-174344-        6792 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 6792)
  AUTHORS   Nakae J, Kitamura T, Silver DL, Accili D
  TITLE     The forkhead transcription factor Foxo1 (Fkhr) confers insulin 
            sensitivity onto glucose-6-phosphatase expression.
  JOURNAL   J Clin Invest. 2001 Nov . 108(9):1359-67.
  PUBMED    11696581
REFERENCE   2  (bases 1 to 6792)
  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: "J Clin 
            Invest. 2001 Nov . 108(9):1359-67."
COMMENT     Sequence Label: HA-Foxo1 (pCMV5)
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..102
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     94..111
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     95..111
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        272..651
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        652..855
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     806..826
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             884..913
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             941..967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3087..3709
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3286..3305)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3733..3753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3738..4067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3918..4053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4105..4124)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4106..4124)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4138..4154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4138..4154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4151..4173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4162..4178
                     /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(4186..4192,4193..4210,4211..4216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4231..4252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4369..4386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4540..5128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4620..4639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5299..6090,6091..6159))
                     /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     5922..5941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6160..6264)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6291..6746
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6378..6397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6588..6609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
       61 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
      121 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
      181 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      241 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
      301 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      361 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      421 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      481 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      541 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      601 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      661 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      721 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      781 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      841 tatataagca gagctcgttt agtgaaccgt cagaattgcc atggagcaga agctgatcag
      901 cgaggaggac ctgaacgaat tcagatctgg taccgccatg tacccatacg atgttccgga
      961 ttacgctgcc gaggcgcccc aggtggtgga gaccgacccg gacttcgagc cgctgccccg
     1021 gcagcgctcc tgtacctggc cgctgcccag gccggagttt aaccagtcca actcgaccac
     1081 ctccagtccg gcgccgtcgg gcggcgcggc cgccaacccc gacgccgcgg cgagcctggc
     1141 ctcggcgtcc gctgtcagca ccgactttat gagcaacctg agcctgctgg aggagagtga
     1201 ggacttcgcg cgggcgccag gctgcgtggc cgtggcggcg gcggctgcgg ccagcagggg
     1261 cctgtgcggg gacttccagg gccccgaggc gggctgcgtg cacccagcgc cgccacagcc
     1321 cccaccgacc gggccgctgt cgcagccccc acccgtgcct ccctccgctg ccgccgccgc
     1381 ggggccactc gcgggacagc cgcgcaagac cagctcgtcg cgccgcaacg cgtggggcaa
     1441 cctgtcgtac gccgacctca tcaccaaggc catcgagagc tcagccgaga agaggctcac
     1501 cctgtcgcag atctacgagt ggatggtgaa gagcgtgccc tacttcaagg ataagggcga
     1561 cagcaacagc tcggcgggct ggaagaattc aattcgccac aatctgtccc ttcacagcag
     1621 gtttattcga gtgcagaatg aaggaactgg aaagagttct tggtggatgc tcaatccaga
     1681 gggaggcaag agcggaaaat caccccggag aagagctgcg tccatggaca acaacagtaa
     1741 atttgctaag agccgagggc gggctgctaa gaaaaaagca tctctccagt ctgggcaaga
     1801 gggtcctgga gacagccctg ggtctcagtt ttctaagtgg cctgcgagtc ctgggtccca
     1861 cagcaacgat gactttgata actggagtac atttcgtcct cgaaccagct caaatgctag
     1921 taccatcagt gggagacttt ctcccatcat gacagagcag gatgacctgg gagatgggga
     1981 cgtgcattcc ctggtgtatc caccctctgc tgccaagatg gcgtctacgc tgcccagtct
     2041 gtctgaaatc agcaatccag aaaacatgga gaaccttctg gataatctca accttctctc
     2101 gtccccaaca tctttaactg tgtccaccca gtcctcgcct ggcagcatga tgcagcagac
     2161 accatgctat tcgtttgcac cgccaaacac cagtctaaat tcacccagtc caaactactc
     2221 aaagtacaca tacggccaat ccagcatgag ccctttgccc cagatgccta tgcagacact
     2281 tcaggacagc aaatcaagtt acggaggatt gaaccagtat aactgtgccc caggactctt
     2341 gaaagagttg ttgacttctg actctcctcc ccacaatgac attatgtcac cggttgatcc
     2401 cggagtggcc caacccaaca gtcgggtcct gggccaaaat gtaatgatgg gccctaattc
     2461 ggtcatgcca gcgtatggca gccaggcatc tcataacaaa atgatgaacc ccagctccca
     2521 cacccaccct ggacatgcac agcaaacggc ttcggtcaac ggccgtaccc tgccccatgt
     2581 ggtgaacacc atgcctcaca catctgccat gaaccgcttg acccccgtga agacaccttt
     2641 acaagtgcct ctgtcccacc ccatgcagat gagtgccctg ggcagctact cctcggtgag
     2701 cagctgcaat ggctatggta ggatgggtgt cctccaccag gagaagctcc caagtgactt
     2761 ggatggcatg tttattgagc gcttggactg tgacatggag tccatcattc ggaatgaccc
     2821 catggatgga gataccttgg attttaactt tgataatgtg ttgcccaacc aaagcttccc
     2881 acacagtgtc aagactacaa cacacagctg ggtgtcaggc taagagttag tgagcaggct
     2941 acatttaaaa gtccttcaga ttgtctgaca gcaggaactg aggagcagtc caaagatgcc
     3001 cttcacccct ccttatagtt ttcaagatta aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3061 gcggccgctg aattctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc
     3121 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     3181 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     3241 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     3301 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     3361 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     3421 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     3481 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     3541 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc
     3601 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg
     3661 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc
     3721 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg
     4081 aactgaaaaa ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc
     4141 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4201 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4261 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4921 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4981 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5161 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     6301 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6361 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6421 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6481 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6541 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6601 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6661 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6721 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa
     6781 gggcgatcgg tg
//
