LOCUS       sequence-429258-        6707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NLS-mAzGreen-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 6707)
  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 6707)
  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: NLS-mAzGreen-Cyclin D1
FEATURES             Location/Qualifiers
     source          1..6707
                     /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             1427..2101
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QAK"
     CDS             1427..2101
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QAK"
     regulatory      2739..2748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3037..3057)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3042..3371
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3222..3357
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3381..3980
                     /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(3381..3400)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3877..3897
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4201..4670
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4800..4817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4971..5559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5051..5070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5719..6510,6511..6579))
                     /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     6342..6361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6580..6684)
                     /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 gaaaggtagg accggctaag tcaccagtga gcgtgatcaa
     1441 gcccgagatg aagatcaagc tgtgcatgag gggcaccgtg aacggccaca acttcgtgat
     1501 cgagggcgag ggcaagggca acccctacga gggcacccag atcctggacc tgaacgtgac
     1561 cgagggcgcc cccctgccct tcgcctacga catcctgacc accgtgttcc agtacggcaa
     1621 cagggccttc accaagtacc ccgccgacat ccaggactac ttcaagcaga ccttccccga
     1681 gggctaccac tgggagagga gcatgaccta cgaggaccag ggcatctgca ccgccaccag
     1741 caacatcagc atgaggggcg actgcttctt ctacgacatc aggttcgacg gcaccaactt
     1801 cccccccaac ggccccgtga tgcagaagaa gaccctgaag tgggagccca gcaccgagaa
     1861 gatgtacgtg gaggacggcg tgctgaaggg cgacgtgaac atgaggctgc tgctggaggg
     1921 cggcggccac tacaggtgcg acttcaagac cacctacaag gccaagaagg aggtgaggct
     1981 gcccgacgcc cacaagatcg accacaggat cgagatcctg aagcacgaca aggactacaa
     2041 caaggtgaag ctgtacgaga acgccgtggc caggtactcc atgctgccca gccaggccaa
     2101 gtccggactc agatctcgag ctcaagcttc gaattcacca gaacaccagc tcctgtgctg
     2161 cgaagtggaa accatccgcc gcgcgtaccc cgatgccaac ctcctcaacg accgggtgct
     2221 gcgggccatg ctgaaggcgg aggagacctg cgcgccctcg gtgtcctact tcaaatgtgt
     2281 gcagaaggag gtcctgccgt ccatgcggaa gatcgtcgcc acctggatgc tggaggtctg
     2341 cgaggaacag aagtgcgagg aggaggtctt cccgctggcc atgaactacc tggaccgctt
     2401 cctgtcgctg gagcccgtga aaaagagccg cctgcagctg ctgggggcca cttgcatgtt
     2461 cgtggcctct aagatgaagg agaccatccc cctgacggcc gagaagctgt gcatctacac
     2521 cgacaactcc atccggcccg aggagctgct gcaaatggag ctgctcctgg tgaacaagct
     2581 caagtggaac ctggccgcaa tgaccccgca cgatttcatt gaacacttcc tctccaaaat
     2641 gccagaggcg gaggagaaca aacagatcat ccgcaaacac gcgcagacct tcgttgccct
     2701 ctgtgccaca gatgtgaagt tcatttccaa tccgccctcc atggtggcag cggggagcgt
     2761 ggtggccgca gtgcaaggcc tgaacctgag gagccccaac aacttcctgt cctactaccg
     2821 cctcacacgc ttcctctcca gagtgatcaa gtgtgaccca gactgcctcc gggcctgcca
     2881 ggagcagatc gaagccctgc tggagtcaag cctgcgccag gcccagcaga acatggaccc
     2941 caaggccgcc gaggaggagg aagaggagga ggaggaggtg gacctggctt gcacacccac
     3001 cgacgtgcgg gacgtggaca tctgagtcga ccctgtggaa tgtgtgtcag ttagggtgtg
     3061 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3121 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3181 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3241 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3301 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3361 gcttttgcaa aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     3421 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     3481 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     3541 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     3601 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     3661 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     3721 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     3781 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     3841 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     3901 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     3961 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     4021 cacgacccca tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4081 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     4141 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     4201 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4261 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4321 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     4381 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     4441 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     4501 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     4561 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     4621 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     4681 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     4741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5401 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5641 tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa gtatatatga
     5701 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5761 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5821 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5881 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5941 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6001 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6061 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6121 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6181 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6241 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6301 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6361 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6421 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6481 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6541 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6601 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6661 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
