LOCUS       sequence-260917-        6036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6036)
  AUTHORS   Di Fiore B, Pines J
  TITLE     Emi1 is needed to couple DNA replication with mitosis but does not 
            regulate activation of the mitotic APC/C.
  JOURNAL   J Cell Biol. 2007 May 7;177(3):425-37.
  PUBMED    17485488
REFERENCE   2  (bases 1 to 6036)
  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: "J Cell 
            Biol."; date: "2007-05-7"; volume: "177(3)"; pages: "425-37"
COMMENT     Sequence Label: pT7-Venus-Emi1 (914)
FEATURES             Location/Qualifiers
     source          1..6036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(607..609,610..612,613..1323)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(652..673)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(913..932)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1260..1281
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2836..2957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2873..2892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2927..2946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2964..3419)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3101..3122)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3313..3332
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3446..3550
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3575..3595)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3619..3829
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3641..3837
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3688..3823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3872..4666
                     /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             3872..4666
                     /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(3926..3945)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4536..4555
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4854..4873)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4898..4945
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5274..5862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5763..5782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgctagc gctaccggtc
      601 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      661 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      721 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
      781 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
      841 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      901 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      961 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1021 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     1081 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1141 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1201 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1261 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1321 aagtccggac tcagatctcg agctcaagct tcgaatttcg agatgagccg gcgcccctgc
     1381 agctgcgccc tacggccacc ccgctgctcc tgcagcgcca gccccagcgc agtgacagcc
     1441 gccgggcgcc ctcgaccctc ggatagttgt aaagaagaaa gttctaccct ttctgtcaaa
     1501 atgaagtgtg attttaattg taaccatgtt cattccggac ttaaactggt aaaacctgat
     1561 gacattggaa gactagtttc ctacacccct gcatatttgg aaggttcctg taaagactgc
     1621 attaaagact atgaaaggct gtcatgtatt gggtcaccga ttgtgagccc taggattgta
     1681 gaacttgaaa ctgaaagcaa gcgcttgcat aacaaggaaa atcaacatgt gcaacagaca
     1741 cttaatagta caaatgaaat agaagcacta gagaccagta gactttatga agacagtggc
     1801 tattcctcat tttctctaca aagtggcctc agtgaacatg aagaaggtag cctcctggag
     1861 gagaatttcg gtgacagtct acaatcctgc ctgctacaaa tacaaagccc agaccaatat
     1921 cccaacaaaa acttgctgcc agttcttcat tttgaaaaag tggtttgttc aacattaaaa
     1981 aagaatgcaa aacgaaatcc taaagtagat cgggagatgc tgaaggaaat tatagccaga
     2041 ggaaatttta gactgcagaa tataattggc agaaaaatgg gcctagaatg tgtagatatt
     2101 ctcagcgaac tctttcgaag gggactcaga catgtcttag caactatttt agcacaactc
     2161 agtgacatgg acttaatcaa tgtgtctaaa gtgagcacaa cttggaagaa gatcctagaa
     2221 gatgataagg gggcattcca gttgtacagt aaagcaatac aaagagttac cgaaaacaac
     2281 aataaatttt cacctcatgc ttcaaccaga gaatatgtta tgttcagaac cccactggct
     2341 tctgttcaga aatcagcagc ccagacttct ctcaaaaaag atgctcaaac caagttatcc
     2401 aatcaaggtg atcagaaagg ttctacttat agtcgacaca atgaattctc tgaggttgcc
     2461 aagacattga aaaagaacga aagcctcaaa gcctgtattc gctgtaattc acctgcaaaa
     2521 tatgattgct atttacaacg ggcaacctgc aaacgagaag gctgtggatt tgattattgt
     2581 acgaagtgtc tctgtaatta tcatactact aaagactgtt cagatggcaa gctcctcaaa
     2641 gccagttgta aaataggtcc cctgcctggt acaaagaaaa gcaaaaagaa tttacgaaga
     2701 ttgtgaggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     2761 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2821 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2881 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2941 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3001 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3061 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3121 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3181 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3241 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3301 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3361 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3421 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3481 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3541 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3601 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3661 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3721 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3781 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3841 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3901 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3961 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4021 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4081 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4141 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4201 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4261 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4321 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4381 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4441 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4501 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4561 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4621 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4681 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4741 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4801 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4861 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4921 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4981 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5041 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5101 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5161 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5221 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5401 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5581 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5641 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5821 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5881 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5941 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     6001 ggggtcatta gttcatagcc catatatgga gttccg
//
