LOCUS       sequence-244388-        6132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  APEX2 fused to the N-terminus of alpha tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6132)
  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 6132)
  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 - Tubulin in pEGFP
FEATURES             Location/Qualifiers
     source          1..6132
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2920..3041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2957..2976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3011..3030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3048..3503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3397..3416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3530..3634
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3636..3993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3636..3827
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3659..3679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3844..3979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4028..4822
                     /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             4028..4822
                     /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(4082..4101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4692..4711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5010..5029)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5054..5101
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5430..6018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5919..5938
                     /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 ccggtcgcca ccatggacta caaggatgac gacgataagg ggaaatcata cccaacagtg
      661 tccgcagact accaggatgc cgtggagaaa gccaagaaga agctgagagg gtttatcgca
      721 gaaaagaggt gcgcacctct gatgctgaga ctggctttcc acagcgcagg cacctttgac
      781 aagggaacca aaacaggcgg accctttgga acaatcaagc accctgctga actggcacat
      841 agtgccaaca atgggctgga catcgccgtg cggctgctgg aacctctgaa agcagagttc
      901 ccaattctgt cctacgccga tttttatcag ctggcaggag tggtcgctgt ggaggtcact
      961 gggggcccca aggtgccttt ccacccagga cgggaggaca aaccagaacc acctccagag
     1021 gggcgcctgc cagatccgac aaagggctcc gaccatctgc gagatgtgtt tgggaaagct
     1081 atgggcctga ctgaccagga tatcgtcgca ctgtctggag ggcacaccat tggcgccgct
     1141 cataaggaaa ggtcaggctt cgagggaccc tggacaagca accccctgat tttcgacaat
     1201 tcttacttta ctgaactgct gagtggagag aaggaagggc tgctgcagct gcccagcgac
     1261 aaagccctgc tgtccgatcc cgtgttcaga cctctggtcg ataagtatgc agccgacgag
     1321 gatgcttttt tcgcagatta cgcagaagca catcagaagc tgtcagaact gggatttgcc
     1381 gacgccaagg gctcgggctc gacctcgggc tcgggctccg gactcagatc tcgagtgcgt
     1441 gagtgcatct ccatccacgt tggccaggct ggtgtccaga ttggcaatgc ctgctgggag
     1501 ctctactgcc tggaacacgg catccagccc gatggccaga tgccaagtga caagaccatt
     1561 gggggaggag atgactcctt caacaccttc ttcagtgaga cgggcgctgg caagcacgtg
     1621 ccccgggctg tgtttgtaga cttggaaccc acagtcattg atgaagttcg cactggcacc
     1681 taccgccagc tcttccaccc tgagcagctc atcacaggca aggaagatgc tgccaataac
     1741 tatgcccgag ggcactacac cattggcaag gagatcattg accttgtgtt ggaccgaatt
     1801 cgcaagctgg ctgaccagtg caccggtctt cagggcttct tggttttcca cagctttggt
     1861 gggggaactg gttctgggtt cacctccctg ctcatggaac gtctctcagt tgattatggc
     1921 aagaagtcca agctggagtt ctccatttac ccagcacccc aggtttccac agctgtagtt
     1981 gagccctaca actccatcct caccacccac accaccctgg agcactctga ttgtgccttc
     2041 atggtagaca atgaggccat ctatgacatc tgtcgtagaa acctcgatat cgagcgccca
     2101 acctacacta accttaaccg ccttattagc cagattgtgt cctccatcac tgcttccctg
     2161 agatttgatg gagccctgaa tgttgacctg acagaattcc agaccaacct ggtgccctac
     2221 ccccgcatcc acttccctct ggccacatat gcccctgtca tctctgctga gaaagcctac
     2281 catgaacagc tttctgtagc agagatcacc aatgcttgct ttgagccagc caaccagatg
     2341 gtgaaatgtg accctcgcca tggtaaatac atggcttgct gcctgttgta ccgtggtgac
     2401 gtggttccca aagatgtcaa tgctgccatt gccaccatca aaaccaagcg cagcatccag
     2461 tttgtggatt ggtgccccac tggcttcaag gttggcatca actaccagcc tcccactgtg
     2521 gtgcctggtg gagacctggc caaggtacag agagctgtgt gcatgctgag caacaccaca
     2581 gccattgctg aggcctgggc tcgcctggac cacaagtttg acctgatgta tgccaagcgt
     2641 gcctttgttc actggtacgt gggtgagggg atggaggaag gcgagttttc agaggcccgt
     2701 gaagatatgg ctgcccttga gaaggattat gaggaggttg gtgtggattc tgttgaagga
     2761 gagggtgagg aagaaggaga ggaatactaa ggatccaccg gatctagata actgatcata
     2821 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2881 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2941 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3001 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3061 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3121 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3181 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3241 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3301 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3361 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3421 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3481 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3601 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3721 agcatgcatc tcaattagtc agcaaccagg kgyggaaagt ccccaggctc cccagcaggc
     3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3841 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3901 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3961 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4021 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4201 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4501 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4681 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4801 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4861 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4921 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4981 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5041 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5101 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5161 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5221 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5281 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6121 accgccatgc at
//
