LOCUS       sequence-429283-        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 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 6659)
  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 6659)
  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: mAzGreen-Cyclin D1-PuroR
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(320..1111,1112..1180))
                     /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     943..962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1181..1285)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1316..1785
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1850..2049
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2250..2666
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2596..2618
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2634..2650
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2678..2687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2684..3361
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             2684..3361
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     regulatory      3999..4008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4297..4317)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4302..4631
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4482..4617
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4641..5240
                     /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(4641..4660)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5137..5157
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5461..5930
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6060..6077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6231..6659,1..160))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6311..6330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
       61 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      241 atcaaaaagg atcttcacct agatcctttt gcggccgcaa atcaatctaa agtatatatg
      301 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      361 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      421 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      481 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
      541 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
      601 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
      661 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      721 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
      781 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
      841 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
      901 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
      961 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1021 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1081 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1141 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1201 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1261 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa
     1321 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     1381 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     1441 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     1501 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     1561 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     1621 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     1681 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     1741 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
     1801 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca agctggccag
     1861 caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc
     1921 ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     1981 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     2041 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     2101 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     2161 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     2221 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     2281 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     2341 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     2401 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     2461 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     2521 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     2581 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     2641 cagccctcac tccttctcta ggcgccggcc ggatcccgcc accatggtga gcgtgatcaa
     2701 gcccgagatg aagatcaagc tgtgcatgag gggcaccgtg aacggccaca acttcgtgat
     2761 cgagggcgag ggcaagggca acccctacga gggcacccag atcctggacc tgaacgtgac
     2821 cgagggcgcc cccctgccct tcgcctacga catcctgacc accgtgttcc agtacggcaa
     2881 cagggccttc accaagtacc ccgccgacat ccaggactac ttcaagcaga ccttccccga
     2941 gggctaccac tgggagagga gcatgaccta cgaggaccag ggcatctgca ccgccaccag
     3001 caacatcagc atgaggggcg actgcttctt ctacgacatc aggttcgacg gcaccaactt
     3061 cccccccaac ggccccgtga tgcagaagaa gaccctgaag tgggagccca gcaccgagaa
     3121 gatgtacgtg gaggacggcg tgctgaaggg cgacgtgaac atgaggctgc tgctggaggg
     3181 cggcggccac tacaggtgcg acttcaagac cacctacaag gccaagaagg aggtgaggct
     3241 gcccgacgcc cacaagatcg accacaggat cgagatcctg aagcacgaca aggactacaa
     3301 caaggtgaag ctgtacgaga acgccgtggc caggtactcc atgctgccca gccaggccaa
     3361 gtccggactc agatctcgag ctcaagcttc gaattcacca gaacaccagc tcctgtgctg
     3421 cgaagtggaa accatccgcc gcgcgtaccc cgatgccaac ctcctcaacg accgggtgct
     3481 gcgggccatg ctgaaggcgg aggagacctg cgcgccctcg gtgtcctact tcaaatgtgt
     3541 gcagaaggag gtcctgccgt ccatgcggaa gatcgtcgcc acctggatgc tggaggtctg
     3601 cgaggaacag aagtgcgagg aggaggtctt cccgctggcc atgaactacc tggaccgctt
     3661 cctgtcgctg gagcccgtga aaaagagccg cctgcagctg ctgggggcca cttgcatgtt
     3721 cgtggcctct aagatgaagg agaccatccc cctgacggcc gagaagctgt gcatctacac
     3781 cgacaactcc atccggcccg aggagctgct gcaaatggag ctgctcctgg tgaacaagct
     3841 caagtggaac ctggccgcaa tgaccccgca cgatttcatt gaacacttcc tctccaaaat
     3901 gccagaggcg gaggagaaca aacagatcat ccgcaaacac gcgcagacct tcgttgccct
     3961 ctgtgccaca gatgtgaagt tcatttccaa tccgccctcc atggtggcag cggggagcgt
     4021 ggtggccgca gtgcaaggcc tgaacctgag gagccccaac aacttcctgt cctactaccg
     4081 cctcacacgc ttcctctcca gagtgatcaa gtgtgaccca gactgcctcc gggcctgcca
     4141 ggagcagatc gaagccctgc tggagtcaag cctgcgccag gcccagcaga acatggaccc
     4201 caaggccgcc gaggaggagg aagaggagga ggaggaggtg gacctggctt gcacacccac
     4261 cgacgtgcgg gacgtggaca tctgagtcga ccctgtggaa tgtgtgtcag ttagggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4501 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4561 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4621 gcttttgcaa aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4681 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4741 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4801 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4861 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4921 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     4981 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5041 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5101 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5161 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5221 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     5281 cacgacccca tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     5341 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     5401 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     5461 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     5521 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5581 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     5641 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     5701 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     5761 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     5821 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     5881 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     5941 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     6001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6061 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacgg
//
