LOCUS       sequence-428750-        7092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TurboID-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 7092)
  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 7092)
  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-Cyclin D1
FEATURES             Location/Qualifiers
     source          1..7092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(243..260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(414..1002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(494..513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1162..1953,1954..2022))
                     /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(2023..2127)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             2158..2627
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2692..2891
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3092..3508
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3438..3460
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3476..3492
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3558..3584
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3588..3614
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3618..3644
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3651..4607
                     /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      5260..5269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5572..5592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5577..5906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5757..5892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5916..6515
                     /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(5916..5935)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6412..6432
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             join(6736..7092,1..113)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
       61 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag
      121 catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg
      181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1081 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata
     1141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg
     2161 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2221 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2281 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     2341 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     2401 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     2461 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     2521 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     2581 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     2641 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg caagctggcc
     2701 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     2761 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     2821 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
     2881 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     2941 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     3001 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     3061 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     3121 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     3181 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     3241 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     3301 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     3361 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     3421 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     3481 tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt acgtaggaat
     3541 tcgcggccgc caccatgtac ccgtatgatg ttccggatta cgctggctat ccctacgacg
     3601 tgcccgacta tgccgggtac ccctatgacg tcccagacta cgcagctagc aaagacaata
     3661 ctgtgcctct gaagctgatc gctctcctgg ctaatggcga gttccatagt ggcgaacagc
     3721 tgggagaaac cctgggcatg tccagggccg ctatcaacaa gcacattcag actctgcgcg
     3781 actggggcgt ggacgtgttc accgtgcccg gaaagggcta ctctctgccc gagcctatcc
     3841 cgctgctgaa cgctaaacag attctgggac agctggacgg cgggagcgtg gcagtcctgc
     3901 ctgtggtcga ctccaccaat cagtacctgc tggatcgaat cggcgagctg aagagtgggg
     3961 atgcttgcat tgcagaatat cagcaggcag ggagaggaag cagagggagg aaatggttct
     4021 ctccttttgg agctaacctg tacctgagta tgttttggcg cctgaagcgg ggaccagcag
     4081 caatcggcct gggcccggtc atcggaattg tcatggcaga agcgctgcga aagctgggag
     4141 cagacaaggt gcgagtcaaa tggcccaatg acctgtatct gcaggataga aagctggcag
     4201 gcatcctggt ggagctggcc ggaataacag gcgatgctgc acagatcgtc attggcgccg
     4261 ggattaacgt ggctatgagg cgcgtggagg aaagcgtggt caatcagggc tggatcacac
     4321 tgcaggaagc agggattaac ctggacagga atactctggc cgctacgctg atccgagagc
     4381 tgcgggcagc cctggaactg ttcgagcagg aaggcctggc tccatatctg ccacggtggg
     4441 agaagctgga taacttcatc aatagacccg tgaagctgat cattggggac aaagagattt
     4501 tcgggattag ccgggggatt gataaacagg gagccctgct gctggaacag gacggagtta
     4561 tcaaaccctg gatgggcgga gaaatcagtc tgcggtctgc cgaaaagctc gagggaggag
     4621 gaggaagcgg cggaggcgga agcggaggag gaggaagccc agaacaccag ctcctgtgct
     4681 gcgaagtgga aaccatccgc cgcgcgtacc ccgatgccaa cctcctcaac gaccgggtgc
     4741 tgcgggccat gctgaaggcg gaggagacct gcgcgccctc ggtgtcctac ttcaaatgtg
     4801 tgcagaagga ggtcctgccg tccatgcgga agatcgtcgc cacctggatg ctggaggtct
     4861 gcgaggaaca gaagtgcgag gaggaggtct tcccgctggc catgaactac ctggaccgct
     4921 tcctgtcgct ggagcccgtg aaaaagagcc gcctgcagct gctgggggcc acttgcatgt
     4981 tcgtggcctc taagatgaag gagaccatcc ccctgacggc cgagaagctg tgcatctaca
     5041 ccgacaactc catccggccc gaggagctgc tgcaaatgga gctgctcctg gtgaacaagc
     5101 tcaagtggaa cctggccgca atgaccccgc acgatttcat tgaacacttc ctctccaaaa
     5161 tgccagaggc ggaggagaac aaacagatca tccgcaaaca cgcgcagacc ttcgttgccc
     5221 tctgtgccac agatgtgaag ttcatttcca atccgccctc catggtggca gcggggagcg
     5281 tggtggccgc agtgcaaggc ctgaacctga ggagccccaa caacttcctg tcctactacc
     5341 gcctcacacg cttcctctcc agagtgatca agtgtgaccc agactgcctc cgggcctgcc
     5401 aggagcagat cgaagccctg ctggagtcaa gcctgcgcca ggcccagcag aacatggacc
     5461 ccaaggccgc cgaggaggag gaagaggagg aggaggaggt ggacctggct tgcacaccca
     5521 ccgacgtgcg ggacgtggac atctgagttt aaaccgctga gtcgaccctg tggaatgtgt
     5581 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5641 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5701 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5761 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5821 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5881 tttttggagg cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc
     5941 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     6001 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     6061 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     6121 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     6181 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     6241 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     6301 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     6361 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     6421 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     6481 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     6541 cgaccgaaag gagcgcacga ccccatgcat cgataaaata aaagatttta tttagtctcc
     6601 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     6661 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     6721 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     6781 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     6841 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     6901 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     6961 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     7021 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
     7081 actgagtcgc cc
//
