LOCUS       sequence-429220-        5538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mAzGreen-Cyclin D2 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5538)
  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 5538)
  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 D2-NeoR
FEATURES             Location/Qualifiers
     source          1..5538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1290
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             613..1290
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     polyA_signal    2326..2447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2363..2382)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2417..2436
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2454..2909)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2591..2612)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2803..2822
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2936..3040
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3042..3399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3042..3233
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3065..3085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3250..3385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3434..4228
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3434..4228
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3488..3507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4098..4117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4416..4435)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4460..4507
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4836..5424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5325..5344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag cgtgatcaag cccgagatga agatcaagct gtgcatgagg
      661 ggcaccgtga acggccacaa cttcgtgatc gagggcgagg gcaagggcaa cccctacgag
      721 ggcacccaga tcctggacct gaacgtgacc gagggcgccc ccctgccctt cgcctacgac
      781 atcctgacca ccgtgttcca gtacggcaac agggccttca ccaagtaccc cgccgacatc
      841 caggactact tcaagcagac cttccccgag ggctaccact gggagaggag catgacctac
      901 gaggaccagg gcatctgcac cgccaccagc aacatcagca tgaggggcga ctgcttcttc
      961 tacgacatca ggttcgacgg caccaacttc ccccccaacg gccccgtgat gcagaagaag
     1021 accctgaagt gggagcccag caccgagaag atgtacgtgg aggacggcgt gctgaagggc
     1081 gacgtgaaca tgaggctgct gctggagggc ggcggccact acaggtgcga cttcaagacc
     1141 acctacaagg ccaagaagga ggtgaggctg cccgacgccc acaagatcga ccacaggatc
     1201 gagatcctga agcacgacaa ggactacaac aaggtgaagc tgtacgagaa cgccgtggcc
     1261 aggtactcca tgctgcccag ccaggccaag tccggactca gatctcgagc tcaagcttcg
     1321 aattcaccag agctgctgtg ccacgaggtg gacccggtcc gcagggccgt gcgggaccgc
     1381 aacctgctcc gagacgaccg cgtcctgcag aacctgctca ccatcgagga gcgctacctt
     1441 ccgcagtgct cctacttcaa gtgcgtgcag aaggacatcc aaccctacat gcgcagaatg
     1501 gtggccacct ggatgctgga ggtctgtgag gaacagaagt gcgaagaaga ggtcttccct
     1561 ctggccatga attacctgga ccgtttcttg gctggggtcc cgactccgaa gtcccatctg
     1621 caactcctgg gtgctgtctg catgttcctg gcctccaaac tcaaagagac cagcccgctg
     1681 accgcggaga agctgtgcat ttacaccgac aactccatca agcctcagga gctgctggag
     1741 tgggaactgg tggtgctggg gaagttgaag tggaacctgg cagctgtcac tcctcatgac
     1801 ttcattgagc acatcttgcg caagctgccc cagcagcggg agaagctgtc tctgatccgc
     1861 aagcatgctc agaccttcat tgctctgtgt gccacggact ttaagtttgc catgtaccca
     1921 ccgtcgatga tcgcaactgg aagtgtggga gcagccatct gtgggctcca gcaggatgag
     1981 gaagtgagct cgctcacttg tgatgccctg actgagctgc tggctaagat caccaacaca
     2041 gacgtggatt gtctcaaagc ttgccaggag cagattgagg cggtgctcct caatagcctg
     2101 cagcagtacc gtcaggacca acgtgacgga tccaagtcgg aggatgaact ggaccaagcc
     2161 agcaccccta cagacgtgcg ggatatcgac ctgtgaagat ccaccggatc tagataactg
     2221 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2281 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2341 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2401 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     2461 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2521 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2581 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2641 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2701 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2761 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2821 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2881 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2941 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3001 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3301 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3421 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3481 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3541 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3601 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3661 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3721 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3781 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3841 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3901 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3961 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4021 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4081 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4141 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4201 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4261 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4321 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4381 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4441 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4501 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4561 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4621 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4981 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5521 cgtattaccg ccatgcat
//
