LOCUS       sequence-337146-        9367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase Reporter for Human PTEN Enhancer PE - Forward 
            Orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9367)
  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 9367)
  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-hPE(+)-Luc
FEATURES             Location/Qualifiers
     source          1..9367
                     /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"
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 ggtaccctcc tctataaaat gtggatttta
     5041 agtaggctat tatagaggat gatgtgagga ttttaaaaga ccatgcatca aaagtaccta
     5101 agatgaagcc tgatttaaga atgagctcag caaatgctag ctcatttcct tggattagcc
     5161 tttctacact gacccaaggc ttgcaaggca aagcaaaact aaataaaagc aaaatgaagt
     5221 taaaaagccc aaacaaataa aaaccaattt gttaattggg tgttggtgga taaaaagact
     5281 ctgaaagtac atccagtcaa atttgaattt caggtaaact atgaaattat ttgtagtata
     5341 agtatataag tatgtagtgt aagtatccca agcattgcat gggatatact tacactcact
     5401 aaattcttca ctgtttattc aaaattccaa tttaactgga tgtcaggtat ttctatttat
     5461 taaacccggc agcctagaca tggtagaatt tagtaagccc agacatgaca gactcctgaa
     5521 tagccaaggc acagaattca aaagagtcaa atgtgtttcc aacttgctaa ctgcaaaagc
     5581 atcaaggaaa aggttgaccc caccctctct gtctctgcca agcaaagagc ccagatggga
     5641 aaccccaggt gagtgattta gcctcctgtg catactctat gggatcctgg tgcagaggtt
     5701 gctgggaaaa ccgtggcagc ctgccctagg ttttccttcg ggtggggccc tgtggttatg
     5761 gtgtggtgac tctttgtcac ggtgcccctg ccacgccctg cttagtccac ctggtggttg
     5821 ggggtagggg agaagagaag ttacaggcag aaaaaaagga cttatcaatg taacatattt
     5881 cttaggtggt ttgtgcattg ctgtttgggc ttgcaaaact ttaattatta ctttgatgca
     5941 gagtggctgc agattgtcac tgccaggcct gatttaatag cctgcttgca caaagtccga
     6001 cagagattgc ttttgttgca ctttctaaca gctcgtgatg gtggcttttt tacgttaagg
     6061 aggaaactct gtggttaaac tgtagcctag gtatttttag agccgttacc atgttaaccg
     6121 aaatttcgta catttcacct gcaggatatg cagtcgtgct tagggccttc cttaaactct
     6181 aacaaaaaat ataatttaac agttgttaat actgtgctta ggaccaactc catgttatca
     6241 gaaaagctac tgaccaaata ctggactgct aagaatcctt tttttggatg ggtggtactg
     6301 tgccattaga gagtattttt ttttccttga tataaaacga gttgaaggaa cataaccaaa
     6361 aggctaaaat tggttagttc taaaggtcag agcctgaagg cagtgaccca tttgttaatc
     6421 attaagcttt tctaagccgt ctctgacaga attctggctt tctgccactt ctctggtcat
     6481 tttaaactgc aaattcttgt actgtttcag tttttaagta ggtttcaaag gttatgagaa
     6541 aggcataact tgttagctat ttttccttat caaacgtctt tagggttttg gaaggcagag
     6601 ctgttctgtg ttaagagaat cctcacagca tacccagcca tacagcattt aaagatggac
     6661 tcttgggtcc tctgaaaagg gtggatttct agctatcatt cccatgttac ttagtttcca
     6721 tagataagca ggatgtgagg gctgaggaca cccacttagc cattaggtat tagttcccaa
     6781 atcactgcta tctcagggta gttgtctcca ataatcccag cactgaaagt tcaaaaaaca
     6841 ggagcaaaat aaccacaatg tatttatcat cttctgtcat cagttataaa gtggaagtac
     6901 tttaaaatat tttgaatgca tttttgtaaa gcccagtgag gttccagtgc tatgacgttt
     6961 cattgccaga aaactcaatg ccagtttatg ctctagtcca tctgaataaa aagtgatctg
     7021 tacacccaaa acacataaac agaactaaat tttatgtagc caagggacat taagctgcta
     7081 aaacatttgc ttcaatattg aacatggaat agctgtcctc tatttctcaa gggaaaaaat
     7141 atacagtcaa accttgctta tccataaccc aaacttccgg aagccttaag tatttggaac
     7201 tgctccaaac ccagaagagg ggaggagtga gaggaataga gtgggggtgg gagaagtggg
     7261 gagcagagag agacagagaa tgaactagcg gaaagcaagc aagtcagctt ttagaagtag
     7321 acctgaagcc ctagcaacac tgaaaagaga cctggagctt acatctgatt ggctttccag
     7381 ccagaaggaa tagagagctc tgattggctg gtgatgatgt ctccaaataa ccagcctttt
     7441 tcaggaaagc aaatttatgt ggcagaaagg acaagagaca gaagaagcac gcaagaagaa
     7501 aagcgaggag aacagagcaa aaaaaaaaaa aaaaaaaaag gaagattatt tgttacagtg
     7561 gtggagactg agacaaaaat gagaataaca agcctattta cgtaactgct tacagtttac
     7621 aaagtgcttt tacatgtaat cattgatatt gtctttacaa agacctgcag ggctcccaga
     7681 agttaaatgg catgccccag gtcacccagc tagtaagaaa aatagtgcat gtcttctggc
     7741 taaaaatcta acacttttga tatggagcag gtgttgtgga gataatgatt tagagttgtc
     7801 ctgtaggaca gagggaagca aatgctctga cagcagaaaa gaaaatcaga aggcacgtca
     7861 gcctgtagtt tagcgggcaa tgcagacagg agtgaagggc tccctgttct tcacgaccac
     7921 acaccttgaa aacatagata taggaaacat ctctgtgggt atttggaagg ggagatggtt
     7981 gtggtacaaa tgaaacctat gtctagaggg ctcagagggc agtcctgaat atatttccct
     8041 ggggtgtatc acagtcgcct ttcatttaac acgtagaagt tggataccta attattcctt
     8101 ccatctatcc ttgaaagctg ttccttcctc aggcttagtc gacaacataa ccattcaccc
     8161 agttgctcaa gctaagaacc tgtaagtcat ctttgtctta attttctttc catgtttaat
     8221 ttatcagcaa gtcctgtaga ttctacttct aaaacacacc ttgaataagg ctactctgtt
     8281 catctccata atcaccattg ttttctcctt ctgagccact gcaataactt gctatttggt
     8341 ttcaatactt tctggcaccc tgtcttcccc agctcttgcc tccttccctt gaaatccatt
     8401 ctctggacta tgccatcctg cttaaaacat gcctgagaat aattcaaaat ccttcccagg
     8461 atctggcccc agtttaattt gcctcactct gcctaggact cactgttcta gctatggagg
     8521 cctccttttt gttatcagaa cacagcacag tctttccagc tgaagagact taaatatttt
     8581 gctggtccct gtctccatgt tatcacatgg ctagctcttt ctcctatctc atgctatcta
     8641 cacagacagt cctactctca caagctgtgt gagagttccc ttcattctca tagaaccttt
     8701 ttattaactt tctgtgacct ctcaaactga aattacttat gtttttattt gattccttgt
     8761 ttatcatcgt ctttttatat gacttccccc tactagtaat caaactccat taagacaagg
     8821 atcaggtcaa ttgccctcat cactgtatcc ccagagctta caacgtggta cacaggagat
     8881 actcaataac tatgtgtcga atgaataagt gaataaatat cagggaatta acatacccta
     8941 catgttacat agtaccatgc atcagtggag ccatggatat gagacactgt gtttcctgtt
     9001 tgtaacctgt tatttgttaa ttcctgtcag agttgagagc tctgatagta gctgccatca
     9061 tctggggagt tctatggaat cttgccacca tgagttctgg aaaagggaat ctgtttgtgt
     9121 tgggacttga gggatggtat aaattaactc ccaaggttta ttagtagtaa acaattaaaa
     9181 tccaccctga aattggagct ttactaaaga tggctatttt gtagcatgta ggaaattacg
     9241 atattggttt acctcctagg attcaaaaag cattcattct cttttggagc cagaaaagga
     9301 gaggtatgtc tcctaacaga actgtgctgg gttttttttt gttattgttg ttttcttttt
     9361 ttgacag
//
