LOCUS       sequence-257380-        5313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Clathrin Vesicles, Excitation: 507 / 572, Emission: 
            516 / 580.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  AUTHORS   Huang F, Hartwich TM, Rivera-Molina FE, Lin Y, Duim WC, Long JJ, 
            Uchil PD, Myers JR, Baird MA, Mothes W, Davidson MW, Toomre D, 
            Bewersdorf J
  TITLE     Video-rate nanoscopy using sCMOS camera-specific single-molecule 
            localization algorithms.
  JOURNAL   Nat Methods. 2013 Jul;10(7):653-8. doi: 10.1038/nmeth.2488. Epub 
            2013 May 26.
  PUBMED    23708387
REFERENCE   2  (bases 1 to 5313)
  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"; doi: 
            "10.1038/nmeth.2488"; journalName: "Nat Methods"; date: "2013-07"; 
            volume: "10"; issue: "7"; pages: "653-8"
COMMENT     Sequence Label: mEos3.2-Clathrin-15
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..601
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     552..572
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             646..1323
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    2134..2255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2171..2190)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2225..2244
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2262..2717)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2399..2420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2611..2630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2744..2848
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2850..3207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2850..3041
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2873..2893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3058..3193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3242..4036
                     /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             3242..4036
                     /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(3296..3315)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3906..3925
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4224..4243)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4268..4315
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4644..5232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5133..5152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      601 tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatgag tgcgattaag
      661 ccagacatga agatcaaact ccgtatggaa ggcaacgtaa acgggcacca ctttgtgatc
      721 gacggagatg gtacaggcaa gccttttgag ggaaaacaga gtatggatct tgaagtcaaa
      781 gagggcggac ctctgccttt tgcctttgat atcctgacca ctgcattcca ttacggcaac
      841 agggtattcg ccaaatatcc agacaacata caagactatt ttaagcagtc gtttcctaag
      901 gggtattcgt gggaacgaag cttgactttc gaagacgggg gcatttgcaa cgccagaaac
      961 gacataacaa tggaagggga cactttctat aataaagttc gattttatgg taccaacttt
     1021 cccgccaatg gtccagttat gcagaagaag acgctgaaat gggagccctc cactgagaaa
     1081 atgtatgtgc gtgatggagt gctgacgggt gatattgaga tggctttgtt gcttgaagga
     1141 aatgcccatt accgatgtga cttcagaact acttacaaag ctaaggagaa gggtgtcaag
     1201 ttaccaggcg cccactttgt ggaccactgc attgagattt taagccatga caaagattac
     1261 aacaaggtta agctgtatga gcatgctgtt gctcattctg gattgcctga caatgccaga
     1321 cgatccggac tcagatctcg agctcaagct tcgaattctg cagtcgacat ggctgatgac
     1381 tttggcttct tctcgtcgtc ggagagtggt gccccggagg cggcggagga ggacccggcg
     1441 gccgccttcc tggcccagca ggagagcgag attgcaggca tagagaacga cgagggcttc
     1501 ggggcacctg ccggcagcca tgcggccccc gcacagccgg gccccacgag tggggctggt
     1561 tctgaggaca tggggaccac agtcaatgga gatgtgtttc aggaggccaa cggtcctgct
     1621 gatggctacg cagccattgc ccaggctgac aggctgaccc aggagcctga gagcatccgc
     1681 aagtggcgag aggagcagag gaaacggctg caagagctgg atgctgcatc taaggtcacg
     1741 gaacaggaat ggcgggagaa ggccaagaag gacctggagg agtggaacca gcgccagagt
     1801 gaacaagtag agaagaacaa gatcaacaac cgggcatccg aggaggcttt cgtgaaggaa
     1861 tccaaggagg agaccccagg cacagagtgg gagaaggtgg cccagctatg tgacttcaac
     1921 cccaagagca gcaagcagtg caaagatgtg tcccgcctgc gctcggtgct catgtccctg
     1981 aagcagacgc cactgtcccg ctagggatcc accggatcta gataactgat cataatcagc
     2041 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2101 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2161 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2221 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     2281 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2341 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2401 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2461 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2521 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2581 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2641 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2701 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2761 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2821 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2881 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2941 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3001 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3061 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3121 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3181 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3241 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3301 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3361 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3421 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3481 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3541 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3601 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3661 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3721 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3781 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3841 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3901 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3961 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4021 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4081 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4141 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4201 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4261 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4321 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4381 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4441 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5281 tttgctcaca tgttctttcc tgcgttatcc cct
//
