LOCUS       sequence-134975-        6480 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 6480)
  AUTHORS   Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, 
            Arden KC, Blenis J, Greenberg ME
  TITLE     Akt promotes cell survival by phosphorylating and inhibiting a 
            Forkhead transcription factor.
  JOURNAL   Cell 1999 Mar 19;96(6):857-68.
  PUBMED    10102273
REFERENCE   2  (bases 1 to 6480)
  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"; 
            date: "1999-03-19"; volume: "96(6)"; pages: "857-68"
COMMENT     Sequence Label: GST-FOXO3a TM 1-506aa
FEATURES             Location/Qualifiers
     source          1..6480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..140
                     /gene="lacI"
                     /label=lacI promoter
     CDS             141..1223
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(160..179)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1272..1277,1278..1295,1296..1302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1310..1326
                     /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)."
     primer_bind     1334..1350
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1334..1350
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1346..1873
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1366..1383)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1366..1382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1375..1397)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1975..1980,1981..1996,1997..2003)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2011..2027
                     /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)."
     CDS             2050..2703
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2661..2683
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2710..2727
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4322..4344)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4489..4507)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4575..4679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4680..4748,4749..5540)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4898..4917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5711..6299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6200..6219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6453..6470
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
       61 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
      121 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      181 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      241 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      301 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      361 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      421 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      481 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      541 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      601 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      661 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      721 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      781 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      841 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      901 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      961 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     1021 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1081 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1141 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1201 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1261 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     1321 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     1381 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1441 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     1501 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     1561 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     1621 gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc
     1681 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     1741 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc
     1801 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
     1861 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     1921 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
     1981 attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     2041 acagtattca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
     2101 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
     2161 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
     2221 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
     2281 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
     2341 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
     2401 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
     2461 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
     2521 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
     2581 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
     2641 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
     2701 aaatcggatc tggttccgcg tggatccccg aattcggcag aggcaccggc ttccccggcc
     2761 ccgctctctc cgctcgaagt ggagctggac ccggagttcg agccccagag ccgtccgcga
     2821 tcctgtgcgt ggcccctgca aaggccggag ctccaagcga gccctgccaa gccctcgggg
     2881 gagacggccg ctgactccat gatccccgag gaggaggacg atgaagacga cgaggacggc
     2941 gggggacggg ccggctcggc catggcgatc ggcggcggcg gcgggagcgg cacgctgggc
     3001 tccgggctgc tccttgagga ctcggcccgg gtgctggcac ccggagggca agaccccggg
     3061 tctgggccag ccaccgcggc gggcgggctg agcgggggta cacaggcgct gctgcagcct
     3121 cagcaaccgc tgccaccgcc gcagccgggg gcggctgggg gctccgggca gccgaggaaa
     3181 tgttcgtcgc ggcggaacgc ctggggaaac ctgtcctacg cggacctgat cacccgcgcc
     3241 atcgagagct ccccggacaa acggctcact ctgtcccaga tctacgagtg gatggtgcgt
     3301 tgcgtgccct acttcaagga taagggcgac agcaacagct ctgccggctg gaagaactcc
     3361 atccggcaca acctgtcact gcatagtcga ttcatgcggg tccagaatga gggaactggc
     3421 aagagctctt ggtggatcat caaccctgat ggggggaaga gcggaaaagc cccccggcgg
     3481 cgggctgtcg caatggacaa tagcaacaag tataccaaga gccgtggccg cgcagccaag
     3541 aagaaggcag ccctgcagac agcccccgaa tcagctgacg acagtccctc ccagctctcc
     3601 aagtggcctg gcagccccac gtcacgcagc agtgatgagc tggatgcgtg gacggacttc
     3661 cgttcacgca ccaatgctaa cgccagcaca gtcagtggcc gcctgtcgcc catcatggca
     3721 agcacagagt tggatgaagt ccaggacgat gatgcgcctc tctcgcccat gctctacagc
     3781 agctcagcca gcctgtcacc ttcagtaagc aagccgtgca cggtggaact gccacggctg
     3841 actgatatgg caggcaccat gaatctgaat gatgggctga ctgaaaacct catggacgac
     3901 ctgctggata acatcacgct cccgccatcc cagccatcgc ccactggggg actcatgcag
     3961 cggagctcta gcttcccgta taccaccaag ggctcgggcc tgggctcccc aaccagctcc
     4021 tttaacagca cggtgttcgg accttcatct ctgaactccc tacgccagtc tcccatgcag
     4081 accatccaag agaacaagcc agctaccttc tcttccatgt cacactatgg taaccagaca
     4141 ctccaggacc tgctcacttc ggactcactt agccacagcg atgtcatgat gacacagtcg
     4201 gaccccttga tgtctcaggc cagcaccgct gtgtctgccc agaattcccg ggtcgactcg
     4261 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
     4321 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4381 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     4441 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
     4501 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4921 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5221 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5821 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6361 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6421 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
