LOCUS       sequence-66693-e        8597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses siRNA-resistant, FKBP-tagged AP-2 alpha subunit and 
            G418 resistance gene in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8597)
  AUTHORS   Robinson MS, Sahlender DA, Foster SD
  TITLE     Rapid inactivation of proteins by rapamycin-induced rerouting to 
            mitochondria.
  JOURNAL   Dev Cell. 2010 Feb 16;18(2):324-31. doi: 
            10.1016/j.devcel.2009.12.015.
  PUBMED    20159602
REFERENCE   2  (bases 1 to 8597)
  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: 
            "10.1016/j.devcel.2009.12"; journalName: "Dev Cell"; date: 
            "2010-02-16- 16"; volume: "18"; issue: "2"; pages: "324-31"
COMMENT     Sequence Label: pIRES-Ap2a2-FKBP
FEATURES             Location/Qualifiers
     source          1..8597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2929..2931,2932..3252)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             complement(3913..3924)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     intron          4353..4582
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4565..4584
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4640..5214,5215..5217,5218..5226)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4807..4824)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5034..5053
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5249..6052
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5312..5331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5922..5941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6084..6101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6090..6314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6361..6380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6440..6456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6440..6456)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6453..6475)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6464..6480
                     /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(6488..6494,6495..6512,6513..6518))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6533..6554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6671..6688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6842..7430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6922..6941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7601..8392,8393..8461))
                     /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     8224..8243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8462..8566)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgcgatatct gcggcctagc tagcgcttaa ggcctgttaa ccggtcgtac
      961 gtctccggat tcgaattcat gccggccgta tccaaagggg acgggatgcg aggcctagcg
     1021 gtcttcatct ccgacatccg aaactgtaaa agtaaagaag ctgaaataaa gagaataaac
     1081 aaagaactgg caaatattag atccaaattt aaaggtgaca aggctcttga tggctacagt
     1141 aaaaagaagt atgtctgcaa attgctcttc atctttctcc ttggtcatga cattgacttt
     1201 ggacacatgg aagctgtgaa tcttctgagc tcaaacagat acacggaaaa gcagattggc
     1261 taccttttca tctctgtatt ggtgaactcg aatagtgaac tgatccggtt gattaacaat
     1321 gccatcaaga atgacctggc cagccgcaat cctacgttta tgggtctcgc cctgcactgc
     1381 attgccaatg tgggtagccg tgagatggcg gaggccttcg ccggagagat ccccaagatc
     1441 ctcgtggcag gagataccat ggatagtgtg aagcagagcg cagccctgtg cttgttacgc
     1501 ctgtacagga catcacctga cctagttccc atgggcgact ggacatctcg tgtggtacac
     1561 ctcctcaatg accagcactt gggggtagtg actgcagcga ctagcctcat caccaccctg
     1621 gcccagaaga accctgagga gttcaagacc tctgtctcgc tagctgtctc gcgactgagc
     1681 aggattgtga cgtctgcatc aacagatctt caggattata cttactactt tgtcccggct
     1741 ccttggctgt cagtcaagct tctgagactg ctgcagtgct acccaccccc agaccctgcg
     1801 gtgcgtggcc gcctgactga gtgcttggag accatcctga acaaggccca ggagccaccc
     1861 aagtccaaga aggtccagca ttcaaatgcc aagaacgctg tgctgtttga ggccatcagc
     1921 ctgatcatcc accatgacag tgagccaaac ttgctcgtcc gcgcctgcaa ccagctgggc
     1981 cagttcctgc agcaccggga gacgaaccta cgctacctgg ctctggagag tatgtgtacc
     2041 ctcgccagct ctgagttctc ccacgaggcc gtcaagaccc acattgagac ggtcatcaat
     2101 gccttgaaga cggaaagaga cgtgagtgtg aggcagcggg cagtggacct gctctacgcc
     2161 atgtgtgacc ggagcaacgc ccagcagatt gtggctgaga tgctgagcta cctagaaacg
     2221 gctgactact ccatccgaga ggaaattgtg ctgaaggtgg ccatcctggc tgagaagtat
     2281 gcagtggact acacttggta tgtggacacc atcctcaacc tcatccgcat cgcgggcgac
     2341 tatgtgagcg aggaggtgtg gtaccgcgtc atccagattg tcatcaatcg cgatgatgtg
     2401 cagggctatg ctgccaagac ggtatttgag gcattgcagg cgccagcatg tcatgagaac
     2461 ttggtcaaag tgggcggcta catcctgggg gagtttggaa acttgatagc tggggaccca
     2521 cgatccagcc ctctgatcca gttcaacctg ctccactcca agttccactt gtgcagcgtc
     2581 cccactcggg cactcctgct gtccacctac atcaagttcg tgaacctctt tccagaggta
     2641 aaagctacca ttcaggacgt gctgcgcagt gacagccagc tgaagaatgc cgacgtggag
     2701 ctgcagcagc gggctgtgga gtacctgcga ctgagcactg tcgccagcac tgacatcctg
     2761 gcaactgtcc tggaggaaat gccacccttc cctgagcgtg agtcctccat cttggccaag
     2821 ctgaagaaga agaagggccc aagcacagtg actgacctgg aggagaccaa gcgggagcga
     2881 agcatcgatg tgaatggggg ccctgaaccc gccaagaacc tgtacaccat gggagtgcag
     2941 gtggaaacca tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg
     3001 gtgcactaca ccgggatgct tgaagatgga aagaaatttg attcctcccg ggacagaaac
     3061 aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt
     3121 gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt
     3181 gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt
     3241 ctaaaactgg aaggatccat cgatgacatc aatgggggtg tggagcccac ccccagcact
     3301 gtgtcgaccc cctcaccctc cgcggacctc ttagggctgc gggcagcccc tccccctgct
     3361 gcacccccgg ctcccgtagg cgggaacctc ctggtggatg tcttctctga cggccccact
     3421 gcacagccca gcctggggcc cactcctgag gaggccttcc tcagcgagct ggagccccct
     3481 gcccctgaga gccccatggc tttgttggct gacccagctc cagctgctga cccaggtcct
     3541 gaggacatag gccctcccat tccagaagca gatgaactgt tgaataagtt cgtgtgtaag
     3601 aatagtgggg tcttgtttga gaaccagctg ctgcagattg gagtcaagtc tgagttccgg
     3661 cagaacctgg gccgcatgta tctcttctat ggcaacaaga cttctgtgca gttccagaac
     3721 ttcttgccca ccgtggtcca tcctggggac ctccagactc agctggcggt gcagaccaag
     3781 cgtgtggcgg cacaagtgga cggtggcgca caggtgcagc aagtactcaa cattgagtgt
     3841 ctgcgagact tcctgacgcc gccactgttg tcggtgcgct tccggtacgg tggcaccgcc
     3901 cagtccctca ctctgaagct cccagtgacc atcaacaaat tcttccagcc cacagagatg
     3961 gcggcccaag actttttcca gcgctggaag cagctgagcc tccccctgca ggaggcacag
     4021 aaaatcttca aagccaacca cccgatggat gctgaagtta ctaaggccaa gcttctgggg
     4081 tttggctctg ctcttctgga caatgtggat cccaaccctg agaactttgt gggtgctgga
     4141 atcatccaga cgaaggccct gcaggtgggg tgtctgcttc ggctggagcc caatgcccag
     4201 gcccaaatgt accgtctaac cctgcgcacc agcaaagagc ctgtctcccg tcacttgtgt
     4261 gagctgctgg cccagcagtt ctgagcggcc gcatagataa ctgatccagt gtgctggaat
     4321 taattcgctg tctgcgaggg ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     4381 gacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     4441 cgcggtgatg cctttgaggg tggccgcgtc catctggtca gaaaagacaa tctttttgtt
     4501 gtcaagcttg aggtgtggca ggcttgagat ctggccatac acttgagtga caatgacatc
     4561 cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcaggt cgagcatgca
     4621 tctagggcgg ccaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     4681 gcttggaata aggccggtgt gcgtttgtct atatgtgatt ttccaccata ttgccgtctt
     4741 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     4801 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     4861 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     4921 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     4981 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     5041 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     5101 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     5161 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc
     5221 acaacccggg ataattcctg cagccaatat gggatcggcc attgaacaag atggattgca
     5281 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5341 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5401 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5461 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5521 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5581 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5641 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5701 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5761 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca
     5821 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5881 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5941 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6001 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatca
     6061 attctctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     6121 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6181 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6241 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6301 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgagtg cattctagtt
     6361 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6421 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6481 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6541 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7021 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7501 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8581 aagtgccacc tgacgtc
//
