LOCUS       sequence-242395-        5706 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 5706)
  AUTHORS   Trinkle-Mulcahy L, Sleeman JE, Lamond AI
  TITLE     Dynamic targeting of protein phosphatase 1 within the nuclei of 
            living mammalian cells.
  JOURNAL   J Cell Sci. 2001 Dec;114(Pt 23):4219-28.
  PUBMED    11739654
REFERENCE   2  (bases 1 to 5706)
  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 
            Sci."; date: "2001-12"; volume: "114(Pt 23)"; pages: "4219-28"
COMMENT     Sequence Label: pEGFP(N3)-PP1beta
FEATURES             Location/Qualifiers
     source          1..5706
                     /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"
     regulatory      561..570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1587..1596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1593..1595,1596..1598,1599..2312)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1593..1595,1596..1598,1599..2309)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1638..1659)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1899..1918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2246..2267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2435..2556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2472..2491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2526..2545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2563..3018)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2700..2721)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2912..2931
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3045..3149
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3151..3508
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3151..3342
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3174..3194)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3359..3494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3543..4337
                     /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             3543..4337
                     /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(3597..3616)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4207..4226
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4525..4544)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4569..4616
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4945..5532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5433..5452
                     /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 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagcc gccaccatgg cggacgggga gctgaacgtg gacagcctca
      601 tcacccggct gctggaggta cgaggatgtc gtccaggaaa gattgtgcag atgactgaag
      661 cagaagttcg aggcttatgt atcaagtctc gggagatctt tctcagccag cctattcttt
      721 tggaattgga agcaccgctg aaaatttgtg gagatattca tggacaatat acagatttac
      781 tgagattatt tgaatatgga ggtttcccac cagaagccaa ctatcttttc ttaggagatt
      841 atgtggacag aggaaagcag tctttggaaa ccatttgttt gctattggct tataaaatca
      901 aatatccaga gaacttcttt ctcttaagag gaaaccatga gtgtgctagc atcaatcgca
      961 tttatggatt ctatgatgaa tgcaaacgaa gatttaatat taaattgtgg aagaccttca
     1021 ctgattgttt taactgtctg cccatagcag ccattgtgga tgagaagatc ttctgttgtc
     1081 atggaggatt gtcaccagac ctgcaatcta tggagcagat tcggagaatt atgagaccta
     1141 ctgatgtccc tgatacaggt ttgctctgtg atttgctatg gtctgatcca gataaggatg
     1201 tgcaaggctg gggagaaaat gatcgtggtg tttcctttac ttttggagct gatgtagtca
     1261 gtaaatttct gaatcgtcat gatttagatt tgatttgtcg agctcatcag gtggtggaag
     1321 atggatatga attttttgct aaacgacagt tggtaacctt attttcagcc ccaaattact
     1381 gtggcgagtt tgataatgct ggtggaatga tgagtgtgga tgaaactttg atgtgttcat
     1441 ttcagatatt gaaaccatct gaaaagaaag ctaaatacca gtatggtgga ctgaattctg
     1501 gacgtcctgt cactccacct cgaacagcta atccgccgaa gaaaaggctg cagtcgacgg
     1561 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1621 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1681 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1741 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1801 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1861 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1921 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1981 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2041 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2101 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2161 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2221 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2281 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     2341 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2401 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2521 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2581 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2641 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2701 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2761 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2821 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2881 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2941 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3001 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3121 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3181 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3241 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3301 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3361 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3421 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3481 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3541 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3601 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3661 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3721 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3781 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3841 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3901 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3961 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4021 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4081 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4141 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4201 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4261 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4321 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4381 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4441 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4501 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4561 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4621 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4681 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4741 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4801 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4861 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5101 gtagcctagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5161 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5221 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5281 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5341 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5401 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5461 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5521 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5581 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5641 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5701 gttccg
//
