LOCUS       sequence-113682-        6163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  APEX2 fused to the C-terminus human vimentin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6163)
  AUTHORS   Lam SS, Martell JD, Kamer KJ, Deerinck TJ, Ellisman MH, Mootha VK, 
            Ting AY
  TITLE     Directed evolution of APEX2 for electron microscopy and proximity 
            labeling.
  JOURNAL   Nat Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179.
  PUBMED    25419960
REFERENCE   2  (bases 1 to 6163)
  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. 2014 Nov 24. doi: 10.1038/nmeth.3179."
COMMENT     Sequence Label: Vimentin - APEX2 in pECFP
FEATURES             Location/Qualifiers
     source          1..6163
                     /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"
     CDS             2803..2826
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2951..3072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2988..3007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3042..3061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3079..3534)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3216..3237)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3428..3447
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3561..3665
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3667..4024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3667..3858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3690..3710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3875..4010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4059..4853
                     /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             4059..4853
                     /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(4113..4132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4723..4742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5041..5060)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5085..5132
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5461..6049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5950..5969
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcccatgt ccaccaggtc cgtgtcctcg
      661 tcctcctacc gcaggatgtt cggcggcccg ggcaccgcga gccggccgag ctccagccgg
      721 agctacgtga ctacgtccac ccgcacctac agcctgggca gcgcgctgcg ccccagcacc
      781 agccgcagcc tctacgcctc gtccccgggc ggcgtgtatg ccacgcgctc ctctgccgtg
      841 cgcctgcgga gcagcgtgcc cggggtgcgg ctcctgcagg actcggtgga cttctcgctg
      901 gccgacgcca tcaacaccga gttcaagaac acccgcacca acgagaaggt ggagctgcag
      961 gagctgaatg accgcttcgc caactacatc gacaaggtgc gcttcctgga gcagcagaat
     1021 aagatcctgc tggccgagct cgagcagctc aagggccaag gcaagtcgcg cctgggggac
     1081 ctctacgagg aggagatgcg ggagctgcgc cggcaggtgg accagctaac caacgacaaa
     1141 gcccgcgtcg aggtggagcg cgacaacctg gccgaggaca tcatgcgcct ccgggagaaa
     1201 ttgcaggagg agatgcttca gagagaggaa gccgaaaaca ccctgcaatc tttcagacag
     1261 gatgttgaca atgcgtctct ggcacgtctt gaccttgaac gcaaagtgga atctttgcaa
     1321 gaagagattg cctttttgaa gaaactccac gaagaggaaa tccaggagct gcaggctcag
     1381 attcaggaac agcatgtcca aatcgatgtg gatgtttcca agcctgacct cacggctgcc
     1441 ctgcgtgacg tacgtcagca atatgaaagt gtggctgcca agaacctgca ggaggcagaa
     1501 gaatggtaca aatccaagtt tgctgacctc tctgaggctg ccaaccggaa caatgacgcc
     1561 ctgcgccagg caaagcagga gtccactgag taccggagac aggtgcagtc cctcacctgt
     1621 gaagtggatg cccttaaagg aaccaatgag tccctggaac gccagatgcg tgaaatggaa
     1681 gagaactttg ccgttgaagc tgctaactac caagacacta ttggccgcct gcaggatgag
     1741 attcagaata tgaaggagga aatggctcgt caccttcgtg aataccaaga cctgctcaat
     1801 gttaagatgg cccttgacat tgagattgcc acctacagga agctgctgga aggcgaggag
     1861 agcaggattt ctctgcctct tccaaacttt tcctccctga acctgaggga aactaatctg
     1921 gattcactcc ctctggttga tacccactca aaaaggacac ttctgattaa gacggttgaa
     1981 actagagatg gacaggttat caacgaaact tctcagcatc acgatgacct tgaaggggat
     2041 ccaccggtcg ccaccgggaa atcataccca acagtgtccg cagactacca ggatgccgtg
     2101 gagaaagcca agaagaagct gagagggttt atcgcagaaa agaggtgcgc acctctgatg
     2161 ctgagactgg ctttccacag cgcaggcacc tttgacaagg gaaccaaaac aggcggaccc
     2221 tttggaacaa tcaagcaccc tgctgaactg gcacatagtg ccaacaatgg gctggacatc
     2281 gccgtgcggc tgctggaacc tctgaaagca gagttcccaa ttctgtccta cgccgatttt
     2341 tatcagctgg caggagtggt cgctgtggag gtcactgggg gccccaaggt gcctttccac
     2401 ccaggacggg aggacaaacc agaaccacct ccagaggggc gcctgccaga tccgacaaag
     2461 ggctccgacc atctgcgaga tgtgtttggg aaagctatgg gcctgactga ccaggatatc
     2521 gtcgcactgt ctggagggca caccattggc gccgctcata aggaaaggtc aggcttcgag
     2581 ggaccctgga caagcaaccc cctgattttc gacaattctt actttactga actgctgagt
     2641 ggagagaagg aagggctgct gcagctgccc agcgacaaag ccctgctgtc cgatcccgtg
     2701 ttcagacctc tggtcgataa gtatgcagcc gacgaggatg cttttttcgc agattacgca
     2761 gaagcacatc agaagctgtc agaactggga tttgccgacg ccgattataa ggacgatgat
     2821 gacaaataag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2881 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2941 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3001 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3061 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3121 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3181 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3241 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3301 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3361 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3421 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3481 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3541 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3601 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3661 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3781 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3841 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3901 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3961 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4021 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4081 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4141 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4201 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4261 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4321 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4381 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4441 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4501 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4561 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4621 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4681 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4741 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4801 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4861 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4921 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4981 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5041 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5101 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5161 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5221 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5281 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5341 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5401 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5461 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5521 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5581 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5641 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5701 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5761 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5821 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5881 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5941 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6001 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6061 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6121 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
