LOCUS       sequence-429284-        6167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TurboID in mammalian cells from a retroviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6167)
  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 6167)
  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-EV
FEATURES             Location/Qualifiers
     source          1..6167
                     /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"
     primer_bind     complement(2497..2517)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2502..2831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2682..2817
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2841..3440
                     /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(2841..2860)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3337..3357
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3661..4130
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4260..4277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4431..5019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4511..4530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5179..5970,5971..6039))
                     /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(6040..6144)
                     /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 gagtagctta agtttaaacc gctgagtcga ccctgtggaa tgtgtgtcag ttagggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2641 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2701 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2761 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2821 gcttttgcaa aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     2881 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     2941 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     3001 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     3061 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     3121 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     3181 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     3241 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     3301 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     3361 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     3421 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     3481 cacgacccca tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     3541 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     3601 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     3661 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     3721 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     3781 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     3841 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     3901 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     3961 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     4021 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     4081 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     4141 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     4201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4321 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4381 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4441 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4501 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4561 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4621 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4681 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4741 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4801 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4861 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4921 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4981 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5041 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5101 tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa gtatatatga
     5161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
