LOCUS       sequence-429286-        7246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TurboID-13x-Cyclin D1 in mammalian cells from a retroviral
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7246)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 7246)
  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"; journalName: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: TurboID-13x-Cyclin D1
FEATURES             Location/Qualifiers
     source          1..7246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1408..1434
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1438..1464
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1468..1494
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1501..2457
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     regulatory      3278..3287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3576..3596)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3581..3910
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3761..3896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3920..4519
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3920..3939)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4416..4436
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4740..5209
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5339..5356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5510..6098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5590..5609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6258..7049,7050..7118))
                     /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"
     promoter        complement(7119..7223)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
     1381 acgtaggaat tcgcggccgc caccatgtac ccgtatgatg ttccggatta cgctggctat
     1441 ccctacgacg tgcccgacta tgccgggtac ccctatgacg tcccagacta cgcagctagc
     1501 aaagacaata ctgtgcctct gaagctgatc gctctcctgg ctaatggcga gttccatagt
     1561 ggcgaacagc tgggagaaac cctgggcatg tccagggccg ctatcaacaa gcacattcag
     1621 actctgcgcg actggggcgt ggacgtgttc accgtgcccg gaaagggcta ctctctgccc
     1681 gagcctatcc cgctgctgaa cgctaaacag attctgggac agctggacgg cgggagcgtg
     1741 gcagtcctgc ctgtggtcga ctccaccaat cagtacctgc tggatcgaat cggcgagctg
     1801 aagagtgggg atgcttgcat tgcagaatat cagcaggcag ggagaggaag cagagggagg
     1861 aaatggttct ctccttttgg agctaacctg tacctgagta tgttttggcg cctgaagcgg
     1921 ggaccagcag caatcggcct gggcccggtc atcggaattg tcatggcaga agcgctgcga
     1981 aagctgggag cagacaaggt gcgagtcaaa tggcccaatg acctgtatct gcaggataga
     2041 aagctggcag gcatcctggt ggagctggcc ggaataacag gcgatgctgc acagatcgtc
     2101 attggcgccg ggattaacgt ggctatgagg cgcgtggagg aaagcgtggt caatcagggc
     2161 tggatcacac tgcaggaagc agggattaac ctggacagga atactctggc cgctacgctg
     2221 atccgagagc tgcgggcagc cctggaactg ttcgagcagg aaggcctggc tccatatctg
     2281 ccacggtggg agaagctgga taacttcatc aatagacccg tgaagctgat cattggggac
     2341 aaagagattt tcgggattag ccgggggatt gataaacagg gagccctgct gctggaacag
     2401 gacggagtta tcaaaccctg gatgggcgga gaaatcagtc tgcggtctgc cgaaaagctc
     2461 gagggatccg gtggaggcgg gtctggaggc gggggtagtg gcgggggtgg aagcgggggt
     2521 ggaggcgggt cgggtggcgg aggtagcgga ggcggtggaa gtggaggcgg tggaagtggt
     2581 ggaggaggtt ctggtggcgg aggtagcggt ggcggaggta gcggaggcgg tggaagtggt
     2641 ggcggaggta gcggaggcgg tggatcggcg ggtttaaacg aacaccagct cctgtgctgc
     2701 gaagtggaaa ccatccgccg cgcgtacccc gatgccaacc tcctcaacga ccgggtgctg
     2761 cgggccatgc tgaaggcgga ggagacctgc gcgccctcgg tgtcctactt caaatgtgtg
     2821 cagaaggagg tcctgccgtc catgcggaag atcgtcgcca cctggatgct ggaggtctgc
     2881 gaggaacaga agtgcgagga ggaggtcttc ccgctggcca tgaactacct ggaccgcttc
     2941 ctgtcgctgg agcccgtgaa aaagagccgc ctgcagctgc tgggggccac ttgcatgttc
     3001 gtggcctcta agatgaagga gaccatcccc ctgacggccg agaagctgtg catctacacc
     3061 gacaactcca tccggcccga ggagctgctg caaatggagc tgctcctggt gaacaagctc
     3121 aagtggaacc tggccgcaat gaccccgcac gatttcattg aacacttcct ctccaaaatg
     3181 ccagaggcgg aggagaacaa acagatcatc cgcaaacacg cgcagacctt cgttgccctc
     3241 tgtgccacag atgtgaagtt catttccaat ccgccctcca tggtggcagc ggggagcgtg
     3301 gtggccgcag tgcaaggcct gaacctgagg agccccaaca acttcctgtc ctactaccgc
     3361 ctcacacgct tcctctccag agtgatcaag tgtgacccag actgcctccg ggcctgccag
     3421 gagcagatcg aagccctgct ggagtcaagc ctgcgccagg cccagcagaa catggacccc
     3481 aaggccgccg aggaggagga agaggaggag gaggaggtgg acctggcttg cacacccacc
     3541 gacgtgcggg acgtggacat ctgagtcgac cctgtggaat gtgtgtcagt tagggtgtgg
     3601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3661 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3721 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3781 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3841 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3901 cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     3961 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4021 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4081 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4141 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4201 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     4261 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4321 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4381 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4441 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4501 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     4561 acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     4621 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4681 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     4741 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4801 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4861 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4921 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4981 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     5041 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     5101 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     5161 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     5221 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     5281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6181 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
     6241 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6301 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6361 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6421 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6481 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6541 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6601 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6661 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6721 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6781 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6841 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6901 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6961 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7021 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7081 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7141 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7201 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
