LOCUS       sequence-340215-        6136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for the constitutive, near-physiological 
            expression of 2xFLAG-2xSTREP-tagged cyclin D3 and a puromycin 
            resistance marker in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6136)
  AUTHORS   Simoneschi D, Rona G, Zhou N, Jeong YT, Jiang S, Milletti G, Arbini 
            AA, O'Sullivan A, Wang AA, Nithikasem S, Keegan S, Siu Y, 
            Cianfanelli V, Maiani E, Nazio F, Cecconi F, Boccalatte F, Fenyo D, 
            Jones DR, Busino L, Pagano M
  TITLE     CRL4(AMBRA1) is a master regulator of D-type cyclins.
  JOURNAL   Nature. 2021 Apr;592(7856):789-793. doi: 10.1038/s41586-021-03445-y.
            Epub 2021 Apr 14.
  PUBMED    33854235
REFERENCE   2  (bases 1 to 6136)
  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: 
            "10.1038/s41586-021-03445-y"; journalName: "Nature"; date: 
            "2021-04"; volume: "592"; issue: "7856"; pages: "789-793"
COMMENT     Sequence Label: pBABE-2xFLAG-2xSTREP-CCND3-Puro
FEATURES             Location/Qualifiers
     source          1..6136
                     /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             1399..1422
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1438..1461
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1477..1500
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1537..1560
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2466..2486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2471..2800
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2651..2786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2810..3409
                     /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(2810..2829)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3306..3326
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3630..4099
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4229..4246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4400..4988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4480..4499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5148..5939,5940..6008))
                     /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     5771..5790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6009..6113)
                     /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 tcagcatgga ctacaaggac gacgatgaca aggtaccagg atctggggat
     1441 tataaagatg atgatgataa agggtcggcc gccagctgga gccaccctca gttcgagaag
     1501 ggaggaggaa gcggcggagg cagcggagga ggaagctgga gccacccgca gttcgagaaa
     1561 ggagctggat ccagcagcga gctgctgtgt tgcgaaggca cccggcacgc gccccgggcc
     1621 gggccggacc cgcggctgct gggggaccag cgtgtcctgc agagcctgct ccgcctggag
     1681 gagcgctacg taccccgcgc ctcctacttc cagtgcgtgc agcgggagat caagccgcac
     1741 atgcggaaga tgctggctta ctggatgctg gaggtatgtg aggagcagcg ctgtgaggag
     1801 gaagtcttcc ccctggccat gaactacctg gatcgctacc tgtcttgcgt ccccacccga
     1861 aaggcgcagt tgcagctcct gggtgcggtc tgcatgctgc tggcctccaa gctgcgcgag
     1921 accacgcccc tgaccatcga aaaactgtgc atctacaccg accacgctgt ctctccccgc
     1981 cagttgcggg actgggaggt gctggtccta gggaagctca agtgggacct ggctgctgtg
     2041 attgcacatg atttcctggc cttcattctg caccggctct ctctgccccg tgaccgacag
     2101 gccttggtca aaaagcatgc ccagaccttt ttggccctct gtgctacaga ttataccttt
     2161 gccatgtacc cgccatccat gatcgccacg ggcagcattg gggctgcagt gcaaggcctg
     2221 ggtgcctgct ccatgtccgg ggatgagctc acagagctgc tggcagggat cactggcact
     2281 gaagtggact gcctgcgggc ctgtcaggag cagatcgaag ctgcactcag ggagagcctc
     2341 agggaagccg ctcagaccag ctccagccca gcgcccaaag ccccccgggg ctccagcagc
     2401 caagggccca gccagaccag cactcctaca gatgtcacag ccatacacct gtagctcgac
     2461 cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2521 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2581 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2641 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2701 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2761 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttacca tgaccgagta
     2821 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     2881 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     2941 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     3001 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     3061 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     3121 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     3181 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     3241 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     3301 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     3361 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     3421 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa aataaaagat
     3481 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     3541 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
     3601 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     3661 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
     3721 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     3781 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     3841 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     3901 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
     3961 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     4021 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     4081 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa
     4141 gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4201 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4261 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4321 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4381 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4441 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4501 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4561 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4621 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4681 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4741 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4801 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4861 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4921 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc
     5101 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6121 ccccgaaaag tgccac
//
