LOCUS       sequence-438440-        6808 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 6808)
  AUTHORS   Nakae J, Barr V, Accili D
  TITLE     Differential regulation of gene expression by insulin and IGF-1 
            receptors correlates with phosphorylation of a single amino acid 
            residue in the forkhead transcription factor FKHR.
  JOURNAL   EMBO J. 2000 Mar 1. 19(5):989-96.
  PUBMED    10698940
REFERENCE   2  (bases 1 to 6808)
  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: "EMBO J. 
            2000 Mar 1. 19(5):989-96."
COMMENT     Sequence Label: Myc-Foxo1 D256
FEATURES             Location/Qualifiers
     source          1..6808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             613..642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2832..3454
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3031..3050)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3478..3498)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3483..3812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3663..3798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3850..3869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3851..3869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3883..3899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3883..3899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3896..3918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3907..3923
                     /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(3931..3937,3938..3955,3956..3961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3976..3997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4114..4131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4285..4873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4365..4384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5044..5835,5836..5904))
                     /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     5667..5686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5905..6009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6036..6491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6123..6142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6333..6354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6617..6639
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6631..6648
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6632..6648
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgcca tggagcagaa gctgatcagc gaggaggacc tgaacgaatt cagatctggt
      661 accacgcgta tcgataagct tgcatgcctg caggtcgact ctagagtcac catggccgag
      721 gcgccccagg tggtggagac cgacccggac ttcgagccgc tgccccggca gcgctcctgt
      781 acctggccgc tgcccaggcc ggagtttaac cagtccaact cgaccacctc cagtccggcg
      841 ccgtcgggcg gcgcggccgc caaccccgac gccgcggcga gcctggcctc ggcgtccgct
      901 gtcagcaccg actttatgag caacctgagc ctgctggagg agagtgagga cttcgcgcgg
      961 gcgccaggct gcgtggccgt ggcggcggcg gctgcggcca gcaggggcct gtgcggggac
     1021 ttccagggcc ccgaggcggg ctgcgtgcac ccagcgccgc cacagccccc accgaccggg
     1081 ccgctgtcgc agcccccacc cgtgcctccc tccgctgccg ccgccgcggg gccactcgcg
     1141 ggacagccgc gcaagaccag ctcgtcgcgc cgcaacgcgt ggggcaacct gtcgtacgcc
     1201 gacctcatca ccaaggccat cgagagctca gccgagaaga ggctcaccct gtcgcagatc
     1261 tacgagtgga tggtgaagag cgtgccctac ttcaaggata agggcgacag caacagctcg
     1321 gcgggctgga agaattcaat tcgccacaat ctgtcccttc acagcaggtt tattcgagtg
     1381 cagaatgaag gaactagaaa gagttcttgg tggatgctca atccagaggg aggcaagagc
     1441 ggaaaatcac cccggagaag agctgcgtcc atggactaga acagtaaatt tgctaagagc
     1501 cgagggcggg ctgctaagaa aaaagcatct ctccagtctg ggcaagaggg tcctggagac
     1561 agccctgggt ctcagttttc taagtggcct gcgagtcctg ggtcccacag caacgatgac
     1621 tttgataact ggagtacatt tcgtcctcga accagctcaa atgctagtac catcagtggg
     1681 agactttctc ccatcatgac agagcaggat gacctgggag atggggacgt gcattccctg
     1741 gtgtatccac cctctgctgc caagatggcg tctacgctgc ccagtctgtc tgaaatcagc
     1801 aatccagaaa acatggagaa ccttctggat aatctcaacc ttctctcgtc cccaacatct
     1861 ttaactgtgt ccacccagtc ctcgcctggc agcatgatgc agcagacacc atgctattcg
     1921 tttgcaccgc caaacaccag tctaaattca cccagtccaa actactcaaa gtacacatac
     1981 ggccaatcca gcatgagccc tttgccccag atgcctatgc agacacttca ggacagcaaa
     2041 tcaagttacg gaggattgaa ccagtataac tgtgccccag gactcttgaa agagttgttg
     2101 acttctgact ctcctcccca caatgacatt atgtcaccgg ttgatcccgg agtggcccaa
     2161 cccaacagtc gggtcctggg ccaaaatgta atgatgggcc ctaattcggt catgccagcg
     2221 tatggcagcc aggcatctca taacaaaatg atgaacccca gctcccacac ccaccctgga
     2281 catgcacagc aaacggcttc ggtcaacggc cgtaccctgc cccatgtggt gaacaccatg
     2341 cctcacacat ctgccatgaa ccgcttgacc cccgtgaaga cacctttaca agtgcctctg
     2401 tcccacccca tgcagatgag tgccctgggc agctactcct cggtgagcag ctgcaatggc
     2461 tatggtagga tgggtgtcct ccaccaggag aagctcccaa gtgacttgga tggcatgttt
     2521 attgagcgct tggactgtga catggagtcc atcattcgga atgaccccat ggatggagat
     2581 accttggatt ttaactttga taatgtgttg cccaaccaaa gcttcccaca cagtgtcaag
     2641 actacaacac acagctgggt gtcaggctaa gagttagtga gcaggctaca tttaaaagtc
     2701 cttcagattg tctgacagca ggaactgagg agcagtccaa agatgccctt cacccctcct
     2761 tatagttttc aagattaaaa aaaaaaaaaa aaaaaaaaaa aaaaagcggc cgctgaattc
     2821 tagaggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     2881 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2941 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3001 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3061 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3121 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3181 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3241 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3301 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     3361 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     3421 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga
     3481 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3661 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3721 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3781 gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga
     3841 aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt
     3901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4021 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4081 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4201 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4261 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4321 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4381 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4441 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4501 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4621 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4681 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4741 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4801 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     6061 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6121 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6181 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6241 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6301 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6361 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6421 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6481 gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6541 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     6601 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc
     6661 tatacattga atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc
     6721 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     6781 ggctcatgtc caatatgacc gccatgtt
//
