LOCUS       sequence-47555-e        6028 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 6028)
  AUTHORS   Jones SA, Shim SH, He J, Zhuang X
  TITLE     Fast, three-dimensional super-resolution imaging of live cells.
  JOURNAL   Nat Methods. 2011 Jun;8(6):499-508. Epub 2011 May 8.
  PUBMED    21552254
REFERENCE   2  (bases 1 to 6028)
  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: "Nat 
            Methods."; date: "2011-06"; volume: "8(6)"; pages: "499-508. Epub 
            2011 May 8"
COMMENT     Sequence Label: ptdEos-CLC
FEATURES             Location/Qualifiers
     source          1..6028
                     /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      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             600..1277
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MVAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             1323..1997
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     polyA_signal    2816..2937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2853..2872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2907..2926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2944..3399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3081..3102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3293..3312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3426..3530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3532..3889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3532..3723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3555..3575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3740..3875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3924..4718
                     /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             3924..4718
                     /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(3978..3997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4588..4607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4906..4925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4950..4997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5326..5914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5815..5834
                     /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 gctagcacca
      601 tggtggcgat taagccagac atgaagatca acctccgtat ggaaggcaac gtaaacgggc
      661 accactttgt gatcgacgga gatggtacag gcaagccttt tgagggaaaa cagagtatgg
      721 atcttgaagt caaagagggc ggacctctgc cttttgcctt tgatatcctg accactgcat
      781 tccattacgg caacagggta ttcgccgaat atccagacca catacaagac tattttaagc
      841 agtcgtttcc taaggggtat tcgtgggaac gaagcttgac tttcgaagac gggggcattt
      901 gcattgccag aaacgacata acaatggaag gggacacttt ctataataaa gttcgatttc
      961 acggtgtaaa ctttcccgcc aatggtccag ttatgcagaa gaagacgctg aaatgggagc
     1021 cctccactga gaaaatgtat gtgcgtgatg gagtgctgac cggtgatatt cgcatggctt
     1081 tgttgcttga aggaaatgcc cattaccgat gtgacttcag aactacttac aaagctaagg
     1141 agaagggtgt caagttacca ggctaccact ttgtggacca ctgcattgag attttaagcc
     1201 atgacaaaga ttacaacaag gttaagctgt atgagcatgc tgttgctcat tctggattgc
     1261 ctgacaatgc cagacgatct cgaggtcacg gtactggttc tactggttct ggttcttctg
     1321 agagtgcgat taagccagac atgaagatca acctccgtat ggaaggcaac gtaaacgggc
     1381 accactttgt gatcgacgga gatggtacag gcaagccttt tgagggaaaa cagagtatgg
     1441 atcttgaagt caaagagggc ggacctctgc cttttgcctt tgatatcctg accactgcat
     1501 tccattacgg caacagggta ttcgccgaat atccagacca catacaagac tattttaagc
     1561 agtcgtttcc taaggggtat tcgtgggaac gaagcttgac tttcgaagac gggggcattt
     1621 gcattgccag aaacgacata acaatggaag gggacacttt ctataataaa gttcgatttc
     1681 acggtgtaaa ctttcccgcc aatggtccag ttatgcagaa gaagacgctg aaatgggagc
     1741 cctccactga gaaaatgtat gtgcgtgatg gagtgctgac cggtgatatt cgcatggctt
     1801 tgttgcttga aggaaatgcc cattaccgat gtgacttcag aactacttac aaagctaagg
     1861 agaagggtgt caagttacca ggctaccact ttgtggacca ctgcattgag attttaagcc
     1921 atgacaaaga ttacaacaag gttaagctgt atgagcatgc tgttgctcat tctggattgc
     1981 ctgacaatgc cagacgagag aattctgcag tcgacggtac catggccgag ttggatccat
     2041 tcggcgctcc cgccggtgcg cccggtggcc ccgctctagg aaacggggtg gcgggcgccg
     2101 gcgaagaaga cccggccgcg gccttcttgg cgcagcaaga gagtgagatt gcgggcatcg
     2161 agaacgatga ggccttcgcc atcctggacg gcggcgcccc cgggccgcag ccacacggcg
     2221 agccgccggg aggtccggat gctgttgacg gagtgatgaa tggtgaatac taccaggaga
     2281 gcaatggtcc aacagacagt tacgcagcca tttcagaagt ggatcgattg caatcagagc
     2341 ctgaaagtat ccgtaagtgg agagaagagc agacagagcg cctggaagcc ctcgatgcca
     2401 attctcggaa gcaagaagcg gagtggaaag aaaaggcgat aaaggagctg gaagagtggt
     2461 atgcgaggca ggacgagcag ctacagaaga caaaggcaaa caacagggca gcagaagaag
     2521 cctttgtaaa tgacattgac gagtcatccc caggcactga gtgggagcgg gtggcccgcc
     2581 tgtgtgactt taaccccaag tccagcaaac aggccaaaga tgtctctcgc atgcgctcag
     2641 tcctcatctc cctgaagcag gcccccctgg tgcactgacg ggcccgggat ccaccggatc
     2701 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2761 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2821 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2881 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     2941 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3001 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3061 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3121 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3181 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3241 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3301 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3361 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3421 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3481 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3541 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3721 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3781 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3841 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3901 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3961 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4021 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4081 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4141 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4201 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4261 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4321 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4381 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4441 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4501 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4561 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4621 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4681 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4741 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4801 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4861 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4921 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4981 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5041 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5101 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5161 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5221 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5281 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5341 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5401 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5461 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5521 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5581 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5641 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5701 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5761 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5821 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5881 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5941 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6001 tgtggataac cgtattaccg ccatgcat
//
