LOCUS       sequence-337145-        9078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase Reporter for Mouse PTEN Enhancer PE - Reverse 
            Orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9078)
  AUTHORS   Tottone L, Lancho O, Loh JW, Singh A, Kimura S, Roels J, Kuchmiy A, 
            Strubbe S, Lawlor MA, da Silva-Diz V, Luo S, Gachet S, Garcia-Prieto
            CA, Hagelaar R, Esteller M, Meijerink JPP, Soulier J, Taghon T, Van 
            Vlierberghe P, Mullighan CG, Khiabanian H, Rocha PP, Herranz D
  TITLE     A Tumor Suppressor Enhancer of PTEN in T-cell development and 
            leukemia.
  JOURNAL   Blood Cancer Discov. 2021 Jan;2(1):92-109. doi: 
            10.1158/2643-3230.BCD-20-0201. Epub 2020 Nov 24.
  PUBMED    33458694
REFERENCE   2  (bases 1 to 9078)
  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"; doi: 
            "10.1158/2643-3230.BCD-20-0201"; journalName: "Blood Cancer Discov";
            date: "2021-01"; volume: "2"; issue: "1"; pages: "92-109"
COMMENT     Sequence Label: pGL3-Promoter-mPE(-)-Luc
FEATURES             Location/Qualifiers
     source          1..9078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      274..283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             280..1932
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(346..364)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1973..2094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1984..2003)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2038..2057
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2342..2359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2513..3101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2593..2612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3272..4063,4064..4132))
                     /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     3895..3914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4133..4237)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4264..4719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4351..4370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4561..4582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4850..4898
                     /note="synthetic polyadenylation signal"
     misc_feature    4912..5003
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4952..4971
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             complement(6085..6096)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
      301 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
      361 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
      421 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
      481 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
      541 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
      601 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
      661 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
      721 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
      781 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
      841 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
      901 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
      961 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     1021 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     1081 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     1141 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     1201 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     1261 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     1321 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     1381 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     1441 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     1501 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     1561 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     1621 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     1681 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     1741 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     1801 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     1861 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     1921 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     1981 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2041 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2101 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2161 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     2221 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2281 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     2341 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2401 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2461 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2521 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2581 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2641 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2701 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2761 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2821 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2881 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2941 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3001 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3061 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3361 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3421 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3541 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4201 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4261 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4681 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     4741 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     4801 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     4861 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     4921 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     4981 tgcaggtgcc agaacatttc tctatcgata ggtaccgtga tgttgacact tcctgatcca
     5041 atcactcctc aaggcaatac tatatggaga ctaagccttc atcagaaagg catttggaac
     5101 tattttatat ccaaactata atacccagta gaagatcatt tccaaaacca ctatctcagt
     5161 ctttgttgag tgctcaccat gtggccagct cttcctctat atgagagaag ctttcatata
     5221 taattgccca tttataatta tctagtagag acatggaagt tcagagtctt tcccaaagtt
     5281 gtacactgat aagtcaatca acttccactt aaatatggac ttcttagctg ccaagtataa
     5341 gatgtgtggc tttcaataaa tggctttaca cctaggagcc gtggcttttt aacctgtcag
     5401 ttggaaattt ccaatcagtt acagatgttc ctacagcctc agtaccactt aagagaccac
     5461 atggacacat cgtatacatc acaacacata gcatgcagag agcacttaga gctactctag
     5521 ggaagagcaa tcagaagtac gttggaggtg cacttacaca caaaacaaac atagtagttt
     5581 agtccaagct gagggaaaac agtcccaatc cagctggcaa gttgtgtcgt tcacttatga
     5641 gccacgagcc atttgttata aaagagaacc taacatgaat taaccactaa agccacattt
     5701 aaacaggact gatgctgtgg actaggtaac tgagtcaaca gcttagcatc attctccctt
     5761 ggctttggta cttgctaact gtatgacctt ggtcgagtca tgtactcttt ctgagcccag
     5821 caaccttatc cataacatgt gggctctaag tagcatatca cacagaatgt tacgatgatg
     5881 aaaaagacca ctcagcctaa gctgatgcac acctttaatc ccagtacttg ggaggcagag
     5941 gcaaacacat ctctgtgagt tcgaggtcaa gctgacaagt tccaggacag ccaaggttac
     6001 acagagacga tacccctcca aaaagaccat tcattgggca ggagagacgg ctcagtaggt
     6061 aaggtcaatg gctgtttcag cggacctacc ttctattccc aggcctgacc agacctctta
     6121 caaccaccca actcctgttc cagggggtct gatgccctcc tcgggccccg gcaggcacca
     6181 gacacataca tgctgcacag gcacacatgc aggcaagtca tacacataaa atgaaatatt
     6241 aaaaaaatta catgcattaa aagtccataa cacaaaggtg agacggagct cagctgctac
     6301 ttcatctcct cacctctggc tctgtataat gagcggaggt ttaagaggca gggtgaaaca
     6361 agaccaaaca aagccagaac aagccaaagc taaatacctg aagcaaaaag tacagttgag
     6421 catgttgttt gggcactcga aggtgagaag ttaaggacga aggatggaag gatttgacta
     6481 gagctatcag agagaacaca caacacccag atctgaattt cagatgaaca atggatttct
     6541 tcagtatccc aaatattgca ccagacattc ttttactaaa taaattattc attgtttatc
     6601 tgaaattcta gtataaatgg gtgtcagtgt tttcatttac caaaccatgc aacctcgaca
     6661 gcctgatttt tgtaagaaat ctcttaaccc tgtaaggaca ctcggagttc agaagagcct
     6721 gacgtgggca actgcagagc catcctggga aaggtcgcac cctctgccac cctttgtttt
     6781 gtgaagcaag ggcctcatgg ggaaaacctc aagtgagcaa ttttcccttc cctgcaccct
     6841 ccacatgagc ctgctgcaga gaatgctggg gagcctgctg cagcctcctt acccgcttgg
     6901 gtctctgtga cccaggtttt ccttgggtga ggccctgtgg tcacgctata gtgactgttt
     6961 gaacccccac cacaccctgc ttggttgcca gtgggtgggg gcagaagagg tagaaacagg
     7021 gaaagaatgc ttagccgttt tgatcagctg ggttgtgctt tgtcagatgg tctcacaaaa
     7081 ctttaattcc tgctttgatg cctagtggcc gcacagtgtc ctcgccaggc ttgtcttaat
     7141 aggctgcttg catgaagtca gacggatcgc ctttgttgca ctttcgtaca cctcatgaca
     7201 gcggctcttt tacgttaagg aggaaactct gtggttaaac tgtagcctag gtatttttag
     7261 agccgttacc atgttaacca aatttcatac atttcacctg caggatatgc agtaacacca
     7321 aggccttccc taaaagctaa ccaaaaaaaa aaaatgtatt taagcaatta atgcaattaa
     7381 tactgtcttt aggactaaga ccatgttatc tgagaacaga gtcgctggtg ctgacacctt
     7441 ttctcttcta ttttttttct ttttgaatgg atggatggta atgtctttaa gggagtacct
     7501 ttttctttga tacaagtgac ttgaagggcc agaaccaaaa gcttaaagct ggttagttct
     7561 aaaggtcaga gcctgaaggc agcgactcct ttgtgaatca ttaagctttc ctaagctgct
     7621 tctgacagac ttctggcttt gcgccacctg tcttgggtca ttttacactg cgaatcctta
     7681 tgctgttcca gtttttaagt cagtagcaaa ggttatgaga aagacagaac ttgtgttgct
     7741 gttgttttcc ttctcaatcc ggcatcgccc ctaagctttc agagcacaga gccagtttgt
     7801 gtttagggca tcttcacacg ccctactaga gagcatttaa agaagaccca tgggtccaca
     7861 gatgagccag atgtgatgga agggataccc attagcaata aagtgagagt ccccatcact
     7921 actgaccagt gattttcacc actaatccca gcactgaaag ttcaagatag aactaatatc
     7981 tgaggtgcta tcagatatta tcttccatca tcagttaaac tggatgagtt ttgacatact
     8041 tagaattcgc tgtgttagcc caatgaggtt caagtgacag gacacgtcac tgccaagttc
     8101 tatgccagtt tatcttctag tcatctgaat aaaagtcact ctgcaccaaa aaaattctct
     8161 taaacacaac gaaattctgt tttatggagc caagggacat taagctgcta aaacacatgc
     8221 tttcaaaatc gaatgtggaa tagctgtcct gtttctcaag ggaaaaaata cacagttaaa
     8281 ccttgcttat tcataataac ccaaacttcc ggaaggctta agcatttgga attgctccaa
     8341 acccaaaaga ggggaggagg gaaagggaag ggtttggaga gcaaggggat gggggggggg
     8401 gggggatgga gggagagaag gacagaatga accagcagca agcaagccag tttttagaag
     8461 tagacctgaa accctagcaa cactgaaaga agacctggag cttgcatctg attggcttac
     8521 cagccagccg gaatgctgag ctctgattgg ctagtgatga tgtctccaaa taatcagcct
     8581 tttgcaggga agcaaattta tctggcagaa aggacaggag gtagaagaag caggcaagaa
     8641 gcaaagggaa gcgaacaggg caaaatgaat gacaggttat ttgtaacaaa ggtggcagct
     8701 gagacgacaa tgagaacaag aatcactatt tatgtagctg cttacagttt ttacaaacta
     8761 ggctccctga aattactgac actgtcttca gaaggacctt caaggattgc agaagtgaaa
     8821 tgacattccc caggttacac agcgagtaag aaaaaagctg aggtcaccgg tctctaatcc
     8881 agggctgttg acaccacaga gtctctggag agaataattt ggagttatcc tgaggaattg
     8941 agctaatcct ctgatcacag aaaagaccac cagcagccac atcagctaca gtttagtggg
     9001 agatgcagaa agggcgtagg gctcactgcc ctttgctaca cagaggttga aatcatcgct
     9061 caagaaatat ctccatgc
//
