LOCUS       sequence-252937-        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-Foxo1ADA (pCMV5)
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..460
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        461..664
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     615..635
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             693..722
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             750..776
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2896..3518
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3095..3114)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3542..3562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3547..3876
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3727..3862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3914..3933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3915..3933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3947..3963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3947..3963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3960..3982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3971..3987
                     /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(3995..4001,4002..4019,4020..4025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4040..4061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4178..4195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4349..4937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4429..4448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5108..5899,5900..5968))
                     /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     5731..5750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5969..6073)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6100..6555
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6187..6206)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6397..6418
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6681..6703
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6695..6712
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6696..6712
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
       61 tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      121 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      181 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      241 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      301 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
      361 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
      421 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      481 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      541 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
      601 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      661 agctcgttta gtgaaccgtc agaattgcca tggagcagaa gctgatcagc gaggaggacc
      721 tgaacgaatt cagatctggt accgccatgt acccatacga tgttccggat tacgctgccg
      781 aggcgcccca ggtggtggag accgacccgg acttcgagcc gctgccccgg cagcgctcct
      841 gtgcctggcc gctgcccagg ccggagttta accagtccaa ctcgaccacc tccagtccgg
      901 cgccgtcggg cggcgcggcc gccaaccccg acgccgcggc gagcctggcc tcggcgtccg
      961 ctgtcagcac cgactttatg agcaacctga gcctgctgga ggagagtgag gacttcgcgc
     1021 gggcgccagg ctgcgtggcc gtggcggcgg cggctgcggc cagcaggggc ctgtgcgggg
     1081 acttccaggg ccccgaggcg ggctgcgtgc acccagcgcc gccacagccc ccaccgaccg
     1141 ggccgctgtc gcagccccca cccgtgcctc cctccgctgc cgccgccgcg gggccactcg
     1201 cgggacagcc gcgcaagacc agctcgtcgc gccgcaacgc gtggggcaac ctgtcgtacg
     1261 ccgacctcat caccaaggcc atcgagagct cagccgagaa gaggctcacc ctgtcgcaga
     1321 tctacgagtg gatggtgaag agcgtgccct acttcaagga taagggcgac agcaacagct
     1381 cggcgggctg gaagaattca attcgccaca atctgtccct tcacagcagg tttattcgag
     1441 tgcagaatga aggaactgga aagagttctt ggtggatgct caatccagag ggaggcaaga
     1501 gcggaaaatc accccggaga agagctgcgg acatggacaa caacagtaaa tttgctaaga
     1561 gccgagggcg ggctgctaag aaaaaagcat ctctccagtc tgggcaagag ggtcctagag
     1621 acagccctgg gtctcagttt tctaagtggc ctgcgagtcc tgggtcccac agcaacgatg
     1681 actttgataa ctggagtaca tttcgtcctc gaaccagcgc aaatgctagt accatcagtg
     1741 ggagactttc tcccatcatg acagagcagg atgacctggg agatggggac gtgcattccc
     1801 tggtgtatcc accctctgct gccaagatgg cgtctacgct gcccagtctg tctgaaatca
     1861 gcaatccaga aaacatggag aaccttctgg ataatctcaa ccttctctcg tccccaacat
     1921 ctttaactgt gtccacccag tcctcgcctg gcagcatgat gcagcagaca ccatgctatt
     1981 cgtttgcacc gccaaacacc agtctaaatt cacccagtcc aaactactca aagtacacat
     2041 acggccaatc cagcatgagc cctttgcccc agatgcctat gcagacactt caggacagca
     2101 aatcaagtta cggaggattg aaccagtata actgtgcccc aggactcttg aaagagttgt
     2161 tgacttctga ctctcctccc cacaatgaca ttatgtcacc ggttgatccc ggagtggccc
     2221 aacccaacag tcgggtcctg ggccaaaatg taatgatggg ccctaattcg gtcatgccag
     2281 cgtatggcag ccaggcatct cataacaaaa tgatgaaccc cagctcccac acccaccctg
     2341 gacatgcaca gcaaacggct tcggtcaacg gccgtaccct gccccatgtg gtgaacacca
     2401 tgcctcacac atctgccatg aaccgcttga cccccgtgaa gacaccttta caagtgcctc
     2461 tgtcccaccc catgcagatg agtgccctgg gcagctactc ctcggtgagc agctgcaatg
     2521 gctatggtag gatgggtgtc ctccaccagg agaagctccc aagtgacttg gatggcatgt
     2581 ttattgagcg cttggactgt gacatggagt ccatcattcg gaatgacccc atggatggag
     2641 ataccttgga ttttaacttt gataatgtgt tgcccaacca aagcttccca cacagtgtca
     2701 agactacaac acacagctgg gtgtcaggct aagagttagt gagcaggcta catttaaaag
     2761 tccttcagat tgtctgacag caggaactga ggagcagtcc aaagatgccc ttcacccctc
     2821 cttatagttt tcaagattaa aaaaaaaaaa aaaaaaaaaa aaaaaaaang cggccgctga
     2881 attctagagg atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     2941 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     3001 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     3061 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     3121 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     3181 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     3241 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     3301 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     3361 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     3421 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     3481 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg
     3541 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3601 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3661 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3721 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3781 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3841 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcctcgagga actgaaaaac
     3901 cagaaagtta attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc
     3961 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4021 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4081 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4141 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4201 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4261 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4321 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5041 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5101 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5161 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5221 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5281 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5341 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5401 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5461 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5521 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5581 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5641 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5701 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5761 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5821 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5881 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5941 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6001 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6061 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     6121 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6181 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6241 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6301 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6361 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6421 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6481 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6541 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6601 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6661 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     6721 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
     6781 aatcaatatt gg
//
