LOCUS       sequence-230313-        5907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  APEX2 fused to the N-terminus of human beta actin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5907)
  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 5907)
  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: APEX2 - Actin in pEGFP
FEATURES             Location/Qualifiers
     source          1..5907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             674..697
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2753..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2844..2863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2881..3336)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3018..3039)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3230..3249
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3363..3467
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3469..3826
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3469..3660
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3492..3512)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3677..3812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3861..4655
                     /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             3861..4655
                     /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(3915..3934)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4525..4544
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4843..4862)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4887..4934
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5263..5851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5752..5771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggactaca aggatgacga cgataagggg aaatcatacc caacagtgtc
      721 cgcagactac caggatgccg tggagaaagc caagaagaag ctgagagggt ttatcgcaga
      781 aaagaggtgc gcacctctga tgctgagact ggctttccac agcgcaggca cctttgacaa
      841 gggaaccaaa acaggcggac cctttggaac aatcaagcac cctgctgaac tggcacatag
      901 tgccaacaat gggctggaca tcgccgtgcg gctgctggaa cctctgaaag cagagttccc
      961 aattctgtcc tacgccgatt tttatcagct ggcaggagtg gtcgctgtgg aggtcactgg
     1021 gggccccaag gtgcctttcc acccaggacg ggaggacaaa ccagaaccac ctccagaggg
     1081 gcgcctgcca gatccgacaa agggctccga ccatctgcga gatgtgtttg ggaaagctat
     1141 gggcctgact gaccaggata tcgtcgcact gtctggaggg cacaccattg gcgccgctca
     1201 taaggaaagg tcaggcttcg agggaccctg gacaagcaac cccctgattt tcgacaattc
     1261 ttactttact gaactgctga gtggagagaa ggaagggctg ctgcagctgc ccagcgacaa
     1321 agccctgctg tccgatcccg tgttcagacc tctggtcgat aagtatgcag ccgacgagga
     1381 tgcttttttc gcagattacg cagaagcaca tcagaagctg tcagaactgg gatttgccga
     1441 cgccaagggc tcgggctcga cctcgggctc gggctccgga ctcagatctc gagctatgga
     1501 tgatgatatc gccgcgctcg tcgtcgacaa cggctccggc atgtgcaagg ccggcttcgc
     1561 gggcgacgat gccccccggg ccgtcttccc ctccatcgtg gggcgcccca ggcaccaggg
     1621 cgtgatggtg ggcatgggtc agaaggattc ctatgtgggc gacgaggccc agagcaagag
     1681 aggcatcctc accctgaagt accccatcga gcacggcatc gtcaccaact gggacgacat
     1741 ggagaaaatc tggcaccaca ccttctacaa tgagctgcgt gtggctcccg aggagcaccc
     1801 cgtgctgctg accgaggccc ccctgaaccc caaggccaac cgcgagaaga tgacccagat
     1861 catgtttgag accttcaaca ccccagccat gtacgttgct atccaggctg tgctatccct
     1921 gtacgcctct ggccgtacca ctggcatcgt gatggactcc ggtgacgggg tcacccacac
     1981 tgtgcccatc tacgaggggt atgccctccc ccatgccatc ctgcgtctgg acctggctgg
     2041 ccgggacctg actgactacc tcatgaagat cctcaccgag cgcggctaca gcttcaccac
     2101 cacggccgag cgggaaatcg tgcgtgacat taaggagaag ctgtgctacg tcgccctgga
     2161 cttcgagcaa gagatggcca cggctgcttc cagctcctcc ctggagaaga gctacgagct
     2221 gcctgacggc caggtcatca ccattggcaa tgagcggttc cgctgccctg aggcactctt
     2281 ccagccttcc ttcctgggca tggagtcctg tggcatccac gaaactacct tcaactccat
     2341 catgaagtgt gacgtggaca tccgcaaaga cctgtacgcc aacacagtgc tgtctggcgg
     2401 caccaccatg taccctggca ttgccgacag gatgcagaag gagatcactg ccctggcacc
     2461 cagcacaatg aagatcaaga tcattgctcc tcctgagcgc aagtactccg tgtggatcgg
     2521 cggctccatc ctggcctcgc tgtccacctt ccagcagatg tggatcagca agcaggagta
     2581 tgacgagtcc ggcccctcca tcgtccaccg caaatgcttc tagggatcca ccggatctag
     2641 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2701 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2761 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2821 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2881 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2941 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3001 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3061 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3121 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3181 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3241 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3301 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3481 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3781 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3841 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3901 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3961 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4021 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4081 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4141 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4201 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4261 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4321 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4381 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4441 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4501 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4561 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4621 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4681 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4741 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4801 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4861 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4921 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4981 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5041 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5101 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5161 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5881 cctggccttt tgctggcctt ttgctca
//
