LOCUS       sequence-66694-e        8572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses siRNA-resistant, FKBP-tagged AP-1 gamma subunit and 
            G418 resistance gene in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8572)
  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 8572)
  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-Ap1g1-FKBP
FEATURES             Location/Qualifiers
     source          1..8572
                     /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(3321..3323,3324..3644)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     intron          4328..4557
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4540..4559
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4615..5189,5190..5192,5193..5201)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4782..4799)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5009..5028
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5224..6027
                     /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(5287..5306)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5897..5916
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6059..6076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6065..6289
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6336..6355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6415..6431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6415..6431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6428..6450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6439..6455
                     /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(6463..6469,6470..6487,6488..6493))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6508..6529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6646..6663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6817..7405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6897..6916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7576..8367,8368..8436))
                     /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     8199..8218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8437..8541)
                     /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 tcgaattccg cagtcgcctc ctcctcggca aatagtcctc ggcgggggcc
     1021 gagtcctcgg gtcatccggg gtgggagacg gcacgggctg cagtcggagg gacgcggggc
     1081 ggccggcgag agggagccgg ggagcggcgt ctcccgggga agcgcgaggc gtgaggtaga
     1141 ggcgccctcc tcgcccggga tccctccgcc tgcagtcgcg gtgctgaggc cgagggagcc
     1201 gccattttgg atgttcggtt cctgctgcca ctgccgccgc cgcccccgaa gctgctggtt
     1261 tccttcgagg tttcgggccg aggatgccag cccccatcag attgcgggag ctgatccgga
     1321 ccatccggac agcccgaacc caagctgagg aacgagaaat gatccagaaa gaatgtgctg
     1381 caatccggtc ctcatttaga gaagaagaca atacataccg gtgtcggaat gtggcaaagt
     1441 tactatatat gcacatgctg ggctaccctg ctcactttgg acagttggag tgcctcaagc
     1501 taatcgcctc acaaaaattc acagacaaac gcattggcta tttaggagca atgctattat
     1561 tagatgaaag acaagatgtg catcttctca tgaccaactg tatcaagaat gatcttaatc
     1621 atagcacgca gtttgtgcag gggctagcac tttgtaccct cggctgcatg ggctcctcgg
     1681 agatgtgcag agatcttgcg ggagaggtag aaaagcttct gaaaacctcc aactcttact
     1741 taagaaaaaa ggcagcactg tgtgctgttc acgtcatcag gaaggttcct gaacttatgg
     1801 agatgttttt accagcaaca aaaaatttat tgaatgagaa gaaccatggt gtccttcaca
     1861 cgtctgtagt cctcctcaca gagatgtgtg aacggagccc agacatgctc gcacatttca
     1921 gaaagcttgt gccccaatta gttcgtattc taaagaacct catcatgtcc ggatattccc
     1981 cagaacatga tgtctctggt atcagtgacc cctttttgca ggtacgaatt ttgcggttat
     2041 taagaatttt aggacggaat gacgatgatt caagtgaagc tatgaatgat atattagcac
     2101 aggttgccac taatacagag actagtaaaa atgtaggaaa tgctattctg tatgagacgg
     2161 ttttgactat catggatatt aaatcagaga gtggattgcg agttctagcc ataaatattc
     2221 ttggtcgatt cttattgaac aatgacaaaa atattagata tgttgctctg acatctctgt
     2281 tgaagactgt gcagacagat cataacgctg tacagaggca tagaagcaca attgtggact
     2341 gtctgaaaga tctggatgtc tccataaaaa gacgtgcaat ggaactgagt tttgccctgg
     2401 tgaatgggaa taatatccga ggcatgatga aagaattact ttattttctg gattcctgtg
     2461 agccagaatt taaagctgat tgtgcatctg gaatcttcct tgctgcagaa aagtacgcac
     2521 cttccaaacg gtggcatata gatacaatta tgcgtgtctt gacaacggca ggaagttatg
     2581 ttcgcgatga tgcagttcct aacttgattc agttaataac caacagtgtg gagatgcacg
     2641 cttacaccgt ccagcgcctg tacaaggcga ttctcggtga ctattctcaa caacccttgg
     2701 tacaagtggc tgcgtggtgt ataggtgaat atggcgatct tcttgtgtct ggccagtgtg
     2761 aagaggaaga gcctattcag gtgacagaag atgaagtgct ggatatttta gaaagtgtcc
     2821 tgatctctaa tatgtccacc tctgtgacaa gaggctatgc tctcactgcc attatgaagc
     2881 tgtctacccg attcacctgt actgtaaacc gaattaagaa agtggtttcc atctatggga
     2941 gcagcatcga cgtagagctc cagcagaggg cagtagagta caacgcactt tttaagaagt
     3001 atgaccacat gaggtctgcc ctacttgaaa gaatgcctgt catggaaaaa gtgaccacaa
     3061 atggcccttc ggagatcgtg cagacaaatg gagagacaga accagcgccc ctagagacta
     3121 aaccaccacc ctcagggcca caacccacaa gccaggccaa tgatctattg gatttgttgg
     3181 gaggaaatga cataacacct gttatcccaa ctgcacctac aagcaaacca gcatcagctg
     3241 gtggagaact tctcgacttg ctaggagaca tcaccatcga tgtgaatggg ggccctgaac
     3301 ccgccaagaa cctgtacacc atgggagtgc aggtggaaac catctcccca ggagacgggc
     3361 gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg
     3421 gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     3481 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     3541 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     3601 cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaggatcc atcgatctga
     3661 cagagctgga gccccctgcc cctgagagcc ccatggcttt gttggctgac ccagctccag
     3721 ctgctgaccc cggggctcca gcggccgctc ctacccctgc ctcagtgcca cagatatccc
     3781 agcccccctt cttgttggat gggctttcct cacagcctct cttcaatgac atcgctccag
     3841 gcatcccctc catcacagcg tacagtaaga acggcttgaa aatcgagttt acatttgaac
     3901 ggtcaaacac caaccccagc gtaacagtga taacgataca ggcctctaac agcacagagc
     3961 tagacatgac ggactttgtt ttccaggctg cagtaccaaa gacattccag ctgcagcttc
     4021 tgtctcctag cagcagcgtt gtcccagcct ttaatacagg gaccatcaca caagtcatta
     4081 aagttctgaa tccacagaag caacagctgc gaatgcggat caagctcaca tataatcaca
     4141 agggctcggc aatgcaagat ctagcagaag tgaacaattt cccccctcag tcctggcaat
     4201 gaggatgcag caccattctc attcttatcc cactcaatca aaggaattcg gatccgcggc
     4261 cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc gagggccagc
     4321 tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag attgtcagtt
     4381 tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt gagggtggcc
     4441 gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg tggcaggctt
     4501 gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc tccacaggtg
     4561 tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat tccgcccctc
     4621 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     4681 tgtctatatg tgattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4741 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4801 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4861 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4921 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4981 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     5041 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     5101 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     5161 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggataat tcctgcagcc
     5221 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5281 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5341 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5401 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5461 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5521 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5581 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5641 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5701 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5761 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5821 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5881 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5941 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6001 tatcgccttc ttgacgagtt cttctgaggg gatcaattct ctagagctcg ctgatcagcc
     6061 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     6121 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     6181 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     6241 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     6301 gaaagaacca gctggggctc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6361 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6421 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6481 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6541 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8461 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8521 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
