LOCUS       sequence-429305-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NLS-HA-TurboID-PCNA in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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: pBABE-NLS-HA-TurboID-PCNA
FEATURES             Location/Qualifiers
     source          1..7163
                     /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"
     regulatory      1370..1379
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1388..1408
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1421..1447
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1451..1477
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1481..1507
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1514..2470
                     /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(3493..3513)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3498..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3678..3813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3837..4436
                     /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(3837..3856)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4333..4353
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4657..5126
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5256..5273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5427..6015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5507..5526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6175..6966,6967..7035))
                     /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(7036..7140)
                     /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 ccggatcccg ccaccatggg
     1381 agcatcacca aaaaagaaga gaaaggtacc ggtggctacc tacccgtatg atgttccgga
     1441 ttacgctggc tatccctacg acgtgcccga ctatgccggg tacccctatg acgtcccaga
     1501 ctacgcagct agcaaagaca atactgtgcc tctgaagctg atcgctctcc tggctaatgg
     1561 cgagttccat agtggcgaac agctgggaga aaccctgggc atgtccaggg ccgctatcaa
     1621 caagcacatt cagactctgc gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg
     1681 ctactctctg cccgagccta tcccgctgct gaacgctaaa cagattctgg gacagctgga
     1741 cggcgggagc gtggcagtcc tgcctgtggt cgactccacc aatcagtacc tgctggatcg
     1801 aatcggcgag ctgaagagtg gggatgcttg cattgcagaa tatcagcagg cagggagagg
     1861 aagcagaggg aggaaatggt tctctccttt tggagctaac ctgtacctga gtatgttttg
     1921 gcgcctgaag cggggaccag cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc
     1981 agaagcgctg cgaaagctgg gagcagacaa ggtgcgagtc aaatggccca atgacctgta
     2041 tctgcaggat agaaagctgg caggcatcct ggtggagctg gccggaataa caggcgatgc
     2101 tgcacagatc gtcattggcg ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt
     2161 ggtcaatcag ggctggatca cactgcagga agcagggatt aacctggaca ggaatactct
     2221 ggccgctacg ctgatccgag agctgcgggc agccctggaa ctgttcgagc aggaaggcct
     2281 ggctccatat ctgccacggt gggagaagct ggataacttc atcaatagac ccgtgaagct
     2341 gatcattggg gacaaagaga ttttcgggat tagccggggg attgataaac agggagccct
     2401 gctgctggaa caggacggag ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc
     2461 tgccgaaaag ctcgagggat ccggtggagg cgggtctgga ggcgggggta gtggcggggg
     2521 tggaagcggg ggtggaggcg ggtcgggtgg cggaggtagc ggaggcggtg gaagtggagg
     2581 cggtggaagt ggtggaggag gttctggtgg cggaggtagc ggtggcggag gtagcggagg
     2641 cggtggaagt ggtggcggag gtagcggagg cggtggatcg aacgcgtatc gatccatgtt
     2701 cgaggcgcgc ctggtccagg gctccatcct caagaaggtg ttggaggcac tcaaggacct
     2761 catcaacgag gcctgctggg atattagctc cagcggtgta aacctgcaga gcatggactc
     2821 gtcccacgtc tctttggtgc agctcaccct gcggtctgag ggcttcgaca cctaccgctg
     2881 cgaccgcaac ctggccatgg gcgtgaacct caccagtatg tccaaaatac taaaatgcgc
     2941 cggcaatgaa gatatcatta cactaagggc cgaagataac gcggatacct tggcgctagt
     3001 atttgaagca ccaaaccagg agaaagtttc agactatgaa atgaagttga tggatttaga
     3061 tgttgaacaa cttggaattc cagaacagga gtacagctgt gtagtaaaga tgccttctgg
     3121 tgaatttgca cgtatatgcc gagatctcag ccatattgga gatgctgttg taatttcctg
     3181 tgcaaaagac ggagtgaaat tttctgcaag tggagaactt ggaaatggaa acattaaatt
     3241 gtcacagaca agtaatgtcg ataaagagga ggaagctgtt accatagaga tgaatgaacc
     3301 agttcaacta acttttgcac tgaggtacct gaacttcttt acaaaagcca ctccactctc
     3361 ttcaacggtg acactcagta tgtctgcaga tgtacccctt gttgtagagt ataaaattgc
     3421 ggatatggga cacttaaaat actacttggc tcccaagatc gaggatgaag aaggatctta
     3481 ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaccatga
     3841 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     3901 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     3961 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4021 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4081 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4141 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4201 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4261 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     4321 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4381 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc
     4441 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat
     4501 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     4561 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     4621 agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
     4681 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     4741 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4801 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     4861 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     4921 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     4981 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     5041 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     5101 gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt
     5161 ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg
     5221 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5281 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5341 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6121 cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7141 cacatttccc cgaaaagtgc cac
//
