LOCUS       sequence-181928-        5484 bp    DNA     circular SYN 15-APR-2025
DEFINITION  for low level expression of mEos2-PCNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5484)
  AUTHORS   Zessin PJ, Sporbert A, Heilemann M
  TITLE     PCNA appears in two populations of slow and fast diffusion with a 
            constant ratio throughout S-phase in replicating mammalian cells.
  JOURNAL   Sci Rep. 2016 Jan 13;6:18779. doi: 10.1038/srep18779.
  PUBMED    26758689
REFERENCE   2  (bases 1 to 5484)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-01-13"; volume: "6:18779. doi"; pages: " 
            10.1038/srep18779"
COMMENT     Sequence Label: pmEos2-Linker_PCNA-MutC
FEATURES             Location/Qualifiers
     source          1..5484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        770..1073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1074..1277
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1228..1248
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1316..1993
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    2981..3102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3018..3037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3072..3091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3109..3564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3246..3267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3458..3477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3591..3695
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3697..4054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3697..3888
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3720..3740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3905..4040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4089..4883
                     /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             4089..4883
                     /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(4143..4162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4753..4772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5071..5090)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5115..5162
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
      721 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      781 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      841 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      901 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      961 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1021 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1081 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1141 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1201 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     1261 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcggtcg ccaccatgag
     1321 tgcgattaag ccagacatga agatcaaact ccgtatggaa ggcaacgtaa acgggcacca
     1381 ctttgtgatc gacggagatg gtacaggcaa gccttttgag ggaaaacaga gtatggatct
     1441 tgaagtcaaa gagggcggac ctctgccttt tgcctttgat atcctgacca ctgcattcca
     1501 ttacggcaac agggtattcg ccaaatatcc agacaacata caagactatt ttaagcagtc
     1561 gtttcctaag gggtattcgt gggaacgaag cttgactttc gaagacgggg gcatttgcat
     1621 tgccagaaac gacataacaa tggaagggga cactttctat aataaagttc gattttatgg
     1681 taccaacttt cccgccaatg gtccagttat gcagaagaag acgctgaaat gggagccctc
     1741 cactgagaaa atgtatgtgc gtgatggagt gctgacgggt gatattcata tggctttgtt
     1801 gcttgaagga aatgcccatt accgatgtga cttcagaact acttacaaag ctaaggagaa
     1861 gggtgtcaag ttaccaggct accactttgt ggaccactgc attgagattt taagccatga
     1921 caaagattac aacaaggtta agctgtatga gcatgctgtt gctcattctg gattgcctga
     1981 caatgccaga cgaggccgcg gcggcgaggg ccagggccag ggccagggcc ccggcagggg
     2041 ctacgcctac aggagctctc gagctatgtt cgaggcgcgc ctggtccagg gctccatcct
     2101 caagaaggtg ttggaggcac tcaaggacct catcaacgag gcctgctggg atattagctc
     2161 cagcggtgta aacctgcaga gcatggactc gtcccacgtc tctttggtgc agctcaccct
     2221 gcggtctgag ggcttcgaca cctaccgctg cgaccgcaac ctggccatgg gcgtgaacct
     2281 caccagtatg tccaaaatac taaaatgcgc cggcaatgaa gatatcatta cactaagggc
     2341 cgaagataac gcggatacct tggcgctagt atttgaagca ccaaaccagg agaaagtttc
     2401 agactatgaa atgaagttga tggatttaga tgttgaacaa cttggaattc cagaacagga
     2461 gtacagctgt gtagtaaaga tgccttctgg tgaatttgca cgtatatgcc gagatctcag
     2521 ccatattgga gacgccgtag ttatatcttg cgcaaaagac ggagtgaaat tttctgcaag
     2581 tggagaactt ggaaatggaa acattaaatt gtcacagaca agtaatgtcg ataaagagga
     2641 ggaagctgtt accatagaga tgaatgaacc agttcaacta acttttgcac tgaggtacct
     2701 gaacttcttt acaaaagcca ctccactctc ttcaacggtg acactcagta tgtctgcaga
     2761 tgtacccctt gttgtagagt ataaaattgc ggatatggga cacttaaaat actacttggc
     2821 tcccaagatc gaggatgaag aaggatctta gggatccacc ggatctagat aactgatcat
     2881 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2941 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3061 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3121 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3181 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3241 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3301 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3361 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3421 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3481 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3541 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3601 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3661 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3961 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4021 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4081 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4141 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4201 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4261 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4321 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4381 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4441 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4501 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4561 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4621 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4681 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4741 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4801 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4861 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4921 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4981 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5041 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5101 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5161 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5221 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5281 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5341 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5401 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5461 agaccccgta gaaaagatca aagg
//
