LOCUS       sequence-429313-        5547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mAzGreen-Cyclin D3 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5547)
  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 5547)
  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 D3-NeoR
FEATURES             Location/Qualifiers
     source          1..5547
                     /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    2335..2456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2372..2391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2426..2445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2463..2918)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2600..2621)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2812..2831
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2945..3049
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3051..3408
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3051..3242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3074..3094)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3259..3394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3443..4237
                     /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             3443..4237
                     /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(3497..3516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4107..4126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4425..4444)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4469..4516
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4845..5433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5334..5353
                     /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 ttgcgaaggc acccggcacg cgccccgggc cgggccggac
     1381 ccgcggctgc tgggggacca gcgtgtcctg cagagcctgc tccgcctgga ggagcgctac
     1441 gtaccccgcg cctcctactt ccagtgcgtg cagcgggaga tcaagccgca catgcggaag
     1501 atgctggctt actggatgct ggaggtatgt gaggagcagc gctgtgagga ggaagtcttc
     1561 cccctggcca tgaactacct ggatcgctac ctgtcttgcg tccccacccg aaaggcgcag
     1621 ttgcagctcc tgggtgcggt ctgcatgctg ctggcctcca agctgcgcga gaccacgccc
     1681 ctgaccatcg aaaaactgtg catctacacc gaccacgctg tctctccccg ccagttgcgg
     1741 gactgggagg tgctggtcct agggaagctc aagtgggacc tggctgctgt gattgcacat
     1801 gatttcctgg ccttcattct gcaccggctc tctctgcccc gtgaccgaca ggccttggtc
     1861 aaaaagcatg cccagacctt tttggccctc tgtgctacag attatacctt tgccatgtac
     1921 ccgccatcca tgatcgccac gggcagcatt ggggctgcag tgcaaggcct gggtgcctgc
     1981 tccatgtccg gggatgagct cacagagctg ctggcaggga tcactggcac tgaagtggac
     2041 tgcctgcggg cctgtcagga gcagatcgaa gctgcactca gggagagcct cagggaagcc
     2101 gctcagacca gctccagccc agcgcccaaa gccccccggg gctccagcag ccaagggccc
     2161 agccagacca gcactcctac agatgtcaca gccatacacc tgtagggatc caccggatct
     2221 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2281 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2341 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2401 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     2461 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2521 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2581 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2641 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2701 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2761 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2821 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2881 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2941 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3001 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3061 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3241 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3301 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3361 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3421 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3481 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3541 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3601 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3661 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3721 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3781 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3841 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3901 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3961 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4021 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4081 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4141 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4201 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4261 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4321 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4381 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4441 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4501 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4561 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4621 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4681 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4741 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4801 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4861 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4921 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4981 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5041 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5101 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5161 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5221 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5281 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5341 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5401 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5461 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5521 gtggataacc gtattaccgc catgcat
//
