LOCUS       sequence-429309-        5556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mAzGreen-Cyclin D1 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5556)
  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 5556)
  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-NeoR
FEATURES             Location/Qualifiers
     source          1..5556
                     /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"
     regulatory      1928..1937
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2344..2465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2381..2400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2435..2454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2472..2927)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2609..2630)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2821..2840
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2954..3058
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3060..3417
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3060..3251
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3083..3103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3268..3403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3452..4246
                     /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             3452..4246
                     /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(3506..3525)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4116..4135
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4434..4453)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4478..4525
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4854..5442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5343..5362
                     /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 aacaccagct cctgtgctgc gaagtggaaa ccatccgccg cgcgtacccc
     1381 gatgccaacc tcctcaacga ccgggtgctg cgggccatgc tgaaggcgga ggagacctgc
     1441 gcgccctcgg tgtcctactt caaatgtgtg cagaaggagg tcctgccgtc catgcggaag
     1501 atcgtcgcca cctggatgct ggaggtctgc gaggaacaga agtgcgagga ggaggtcttc
     1561 ccgctggcca tgaactacct ggaccgcttc ctgtcgctgg agcccgtgaa aaagagccgc
     1621 ctgcagctgc tgggggccac ttgcatgttc gtggcctcta agatgaagga gaccatcccc
     1681 ctgacggccg agaagctgtg catctacacc gacaactcca tccggcccga ggagctgctg
     1741 caaatggagc tgctcctggt gaacaagctc aagtggaacc tggccgcaat gaccccgcac
     1801 gatttcattg aacacttcct ctccaaaatg ccagaggcgg aggagaacaa acagatcatc
     1861 cgcaaacacg cgcagacctt cgttgccctc tgtgccacag atgtgaagtt catttccaat
     1921 ccgccctcca tggtggcagc ggggagcgtg gtggccgcag tgcaaggcct gaacctgagg
     1981 agccccaaca acttcctgtc ctactaccgc ctcacacgct tcctctccag agtgatcaag
     2041 tgtgacccag actgcctccg ggcctgccag gagcagatcg aagccctgct ggagtcaagc
     2101 ctgcgccagg cccagcagaa catggacccc aaggccgccg aggaggagga agaggaggag
     2161 gaggaggtgg acctggcttg cacacccacc gacgtgcggg acgtggacat ctgaggatcc
     2221 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2281 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2341 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2401 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2461 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2521 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2581 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2641 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2701 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2761 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2821 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2881 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2941 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3001 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3061 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3421 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3481 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3541 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3601 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3661 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3721 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3781 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3841 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3901 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3961 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4021 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4081 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4141 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4201 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4261 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4321 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4381 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4441 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4501 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4561 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4621 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4681 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4741 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4801 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4861 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4921 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4981 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5041 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5101 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5161 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5221 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5281 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5341 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5401 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5461 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5521 cctgattctg tggataaccg tattaccgcc atgcat
//
