LOCUS       sequence-196953-        7078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses hPTEN fused to the N-terminal part of Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7078)
  AUTHORS   Marciniak E, Leboucher A, Caron E, Ahmed T, Tailleux A, Dumont J, 
            Issad T, Gerhardt E, Pagesy P, Vileno M, Bournonville C, Hamdane M, 
            Bantubungi K, Lancel S, Demeyer D, Eddarkaoui S, Vallez E, Vieau D, 
            Humez S, Faivre E, Grenier-Boley B, Outeiro TF, Staels B, Amouyel P,
            Balschun D, Buee L, Blum D
  TITLE     Tau deletion promotes brain insulin resistance.
  JOURNAL   J Exp Med. 2017 Aug 7;214(8):2257-2269. doi: 10.1084/jem.20161731. 
            Epub 2017 Jun 26.
  PUBMED    28652303
REFERENCE   2  (bases 1 to 7078)
  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: 
            "10.1084/jem.20161731"; journalName: "J Exp Med"; date: "2017-08-7- 
            7"; volume: "214"; issue: "8"; pages: "2257-2269"
COMMENT     Sequence Label: VN-PTEN
FEATURES             Location/Qualifiers
     source          1..7078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        231..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        611..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     859..878
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        859..877
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(900..902,903..905,906..1364)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(945..966)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1206..1225)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2665..2682)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2671..2895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2941..3369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3028..3047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3238..3259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3378..3398)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3383..3712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3563..3698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3625..3644
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3779..4573
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3833..3852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4443..4462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4747..4868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4784..4803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4838..4857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4917..4933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4917..4933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4930..4952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4941..4957
                     /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(4965..4971,4972..4989,4990..4995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5010..5031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5148..5165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5319..5907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5399..5418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6078..6869,6870..6938))
                     /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     6701..6720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6939..7043)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
       61 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
      121 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      181 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
      841 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg gctagcacca
      901 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      961 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1021 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1081 tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     1141 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1201 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1261 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1321 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag caaatcgatg
     1381 gtggcggtgg ctctggaggt ggtgggtccc ttaagatgac agccatcatc aaagagatcg
     1441 ttagcagaaa caaaaggaga tatcaagagg atggattcga cttagacttg acctatattt
     1501 atccaaacat tattgctatg ggatttcctg cagaaagact tgaaggcgta tacaggaaca
     1561 atattgatga tgtagtaagg tttttggatt caaagcataa aaaccattac aagatataca
     1621 atctttgtgc tgaaagacat tatgacaccg ccaaatttaa ttgcagagtt gcacaatatc
     1681 cttttgaaga ccataaccca ccacagctag aacttatcaa acccttttgt gaagatcttg
     1741 accaatggct aagtgaagat gacaaccatg ttgcagcaat tcactgtaaa gctggaaagg
     1801 gacgaactgg tgtaatgata tgtgcatatt tattacatcg gggcaaattt ttaaaggcac
     1861 aagaggccct agatttctat ggggaagtaa ggaccagaga caaaaaggga gtaactattc
     1921 ccagtcagag gcgctatgtg tattattata gctacctgtt aaagaatcat ctggattata
     1981 gaccagtggc actgttgttt cacaagatga tgtttgaaac tattccaatg ttcagtggcg
     2041 gaacttgcaa tcctcagttt gtggtctgcc agctaaaggt gaagatatat tcctccaatt
     2101 caggacccac acgacgggaa gacaagttca tgtactttga gttccctcag ccgttacctg
     2161 tgtgtggtga tatcaaagta gagttcttcc acaaacagaa caagatgcta aaaaaggaca
     2221 aaatgtttca cttttgggta aatacattct tcataccagg accagaggaa acctcagaaa
     2281 aagtagaaaa tggaagtcta tgtgatcaag aaatcgatag catttgcagt atagagcgtg
     2341 cagataatga caaggaatat ctagtactta ctttaacaaa aaatgatctt gacaaagcaa
     2401 ataaagacaa agccaaccga tacttttctc caaattttaa ggtgaagctg tacttcacaa
     2461 aaacagtaga ggagccgtca aatccagagg ctagcagttc aacttctgta acaccagatg
     2521 ttagtgacaa tgaacctgat cattatagat attctgacac cactgactct gatccagaga
     2581 atgaaccttt tgatgaagat cagcatacac aaattacaaa agtctgactc gagtctagag
     2641 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2701 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2761 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2821 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2881 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3001 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3061 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3121 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3241 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3301 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3361 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3721 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3781 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3841 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3901 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3961 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4021 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4081 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4141 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4201 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4261 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4321 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4381 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4441 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4501 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4561 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4621 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4681 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4741 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4861 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4981 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5401 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacg
//
