LOCUS       sequence-180925-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the isoform IV of Promyelocytic Leukemia Protein (PML) in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  AUTHORS   Guo L, Giasson BI, Glavis-Bloom A, Brewer MD, Shorter J, Gitler AD, 
            Yang X
  TITLE     A Cellular System that Degrades Misfolded Proteins and Protects 
            against Neurodegeneration.
  JOURNAL   Mol Cell. 2014 Jul 3;55(1):15-30. doi: 10.1016/j.molcel.2014.04.030.
            Epub 2014 May 29.
  PUBMED    24882209
REFERENCE   2  (bases 1 to 6633)
  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.molcel.2014.04.030"; journalName: "Mol Cell"; date: 
            "2014-07-3- 3"; volume: "55"; issue: "1"; pages: "15-30"
COMMENT     Sequence Label: Flag-PML IV/pRK5
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..18
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2..18
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        47..426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        427..630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     581..601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     627..651
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        861..879
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     861..878
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     892..911
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      958..967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             967..990
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2913..2936
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2966..2975
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2979..3113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3016..3035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3070..3089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3182..3539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3182..3373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3205..3225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3390..3525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3559..3576)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3559..3575)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3568..3590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3788..4243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3875..3894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4085..4106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4260..4279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4379..4401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4439..4457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4525..4629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4630..4698,4699..5490)
                     /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     complement(4848..4867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5661..6249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6150..6169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6403..6420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6537..6558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6573..6578,6579..6596,6597..6603)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6611..6627
                     /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)."
     primer_bind     join(6616..6633,1..5)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 acaggaaaca gctatgacat gattacgaat taattcgagc tcgcccgaca ttgattattg
       61 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      661 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccgcggc
      721 cgggaacggt gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat
      781 agagtctata ggcccacccc cttggcttcg ttagaacgcg gctacaatta atacataacc
      841 ttatgtatca tacacatacg atttaggtga cactatagaa taacatccac tttgcctttc
      901 tctccacagg tgtccactcc caggtccaac tgcacctcgg ttctatcgat tgaattcgcc
      961 gccatggact acaaggacga cgatgacaag ggatccacca tggagcctgc acccgcccga
     1021 tctccgaggc cccagcagga ccccgcccgg ccccaggagc ccaccatgcc tccccccgag
     1081 accccctctg aaggccgcca gcccagcccc agccccagcc ctacagagcg agcccccgct
     1141 tcggaggagg agttccagtt tctgcgctgc cagcaatgcc aggcggaagc caagtgcccg
     1201 aagctgctgc cttgtctgca cacgctgtgc tcaggatgcc tggaggcgtc gggcatgcag
     1261 tgccccatct gccaggcgcc ctggccccta ggtgcagaca cacccgccct ggataacgtc
     1321 tttttcgaga gtctgcagcg gcgcctgtcg gtgtaccggc agattgtgga tgcgcaggct
     1381 gtgtgcaccc gctgcaaaga gtcggccgac ttctggtgct ttgagtgcga gcagctcctc
     1441 tgcgccaagt gcttcgaggc acaccagtgg ttcctcaagc acgaggcccg gcccctagca
     1501 gagctgcgca accagtcggt gcgtgagttc ctggacggca cccgcaagac caacaacatc
     1561 ttctgctcca accccaacca ccgcacccct acgctgacca gcatctactg ccgaggatgt
     1621 tccaagccgc tgtgctgctc gtgcgcgctc cttgacagca gccacagtga gctcaagtgc
     1681 gacatcagcg cagagatcca gcagcgacag gaggagctgg acgccatgac gcaggcgctg
     1741 caggagcagg atagtgcctt tggcgcggtt cacgcgcaga tgcacgcggc cgtcggccag
     1801 ctgggccgcg cgcgtgccga gaccgaggag ctgatccgcg agcgcgtgcg ccaggtggta
     1861 gctcacgtgc gggctcagga gcgcgagctg ctggaggctg tggacgcgcg gtaccagcgc
     1921 gactacgagg agatggccag tcggctgggc cgcctggatg ctgtgctgca gcgcatccgc
     1981 acgggcagcg cgctggtgca gaggatgaag tgctacgcct cggaccagga ggtgctggac
     2041 atgcacggtt tcctgcgcca ggcgctctgc cgcctgcgcc aggaggagcc ccagagcctg
     2101 caagctgccg tgcgcaccga tggcttcgac gagttcaagg tgcgcctgca ggacctcagc
     2161 tcttgcatca cccaggggaa agatgcagct gtatccaaga aagccagccc agaggctgcc
     2221 agcactccca gggaccctat tgacgttgac ctgcccgagg aggcagagag agtgaaggcc
     2281 caggttcagg ccctggggct ggctgaagcc cagcctatgg ctgtggtaca gtcagtgccc
     2341 ggggcacacc ccgtgccagt gtacgccttc tccatcaaag gcccttccta tggagaggat
     2401 gtctccaata caacgacagc ccagaagagg aagtgcagcc agacccagtg ccccaggaag
     2461 gtcatcaaga tggagtctga ggaggggaag gaggcaaggt tggctcggag ctccccggag
     2521 cagcccaggc ccagcacctc caaggcagtc tcaccacccc acctggatgg accgcctagc
     2581 cccaggagcc ccgtcatagg aagtgaggtc ttcctgccca acagcaacca cgtggccagt
     2641 ggcgccgggg aggcagagga acgcgttgtg gtgatcagca gctcggaaga ctcagatgcc
     2701 gaaaactcgt cctcccgaga gctggatgac agcagcagtg agtccagtga cctccagctg
     2761 gaaggcccca gcaccctcag ggtcctggac gagaaccttg ctgaccccca agcagaagac
     2821 agacctctgg ttttctttga cctcaagatt gacaatgaaa gtgggttctc ctggggctac
     2881 ccccacccct ttctaattta ggtcgacgac tcgactacaa ggacgacgat gacaagtagg
     2941 cggccgctcg acctgcagaa gcttggccgc catggcccaa cttgtttatt gcagcttata
     3001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3061 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcgggaatt
     3121 aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
     3181 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3541 agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3601 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3661 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     3721 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3781 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3841 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3901 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3961 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4021 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4081 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     4141 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4201 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4261 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4321 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4381 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4441 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4501 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4621 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4681 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4741 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4801 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4861 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4921 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4981 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5041 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5101 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5161 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5221 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5281 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5341 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5401 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5461 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6361 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6421 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6481 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6541 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6601 ttgtgtggaa ttgtgagcgg ataacaattt cac
//
