LOCUS       sequence-400018-        7826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gamma-retroviral vector that expresses mNeonGreen fused to APEX3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7826)
  AUTHORS   Becker JT, Auerbach AA, Harris RS
  TITLE     APEX3 - an optimized tool for rapid and unbiased proximity labeling.
  JOURNAL   J Mol Biol. 2023 May 12:168145. doi: 10.1016/j.jmb.2023.168145.
  PUBMED    37182813
REFERENCE   2  (bases 1 to 7826)
  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: "J Mol Biol.
            2023 May 12:168145. doi: 10.1016/j.jmb.2023.168145."
COMMENT     Sequence Label: mNeonGreen-APEX3
FEATURES             Location/Qualifiers
     source          1..7826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    165..506
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             573..989
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     919..941
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     957..973
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1004..1013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1010..1717
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     CDS             1772..2518
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKASELGFADA"
     CDS             2534..2575
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2771..2788)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2998..3017
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             3805..4319
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4481..4497
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4505..4511,4512..4529,4530..4535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4550..4571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4688..4705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4859..5447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4939..4958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5618..6409,6410..6478))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6241..6260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6479..6583)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6651..6669
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6707..6729)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6829..6848
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7042..7064
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7192..7211
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7411..7826,1..101)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
       61 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
      121 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
      181 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
      241 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
      301 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
      361 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
      421 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
      481 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
      541 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
      601 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
      661 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
      721 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
      781 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
      841 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
      901 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
      961 atccagccct cactccttct ctaggcgcct gtatgaattc gccgccacca tggtgagcaa
     1021 gggcgaggag gataacatgg cctctctccc agcgacacat gagttacaca tctttggctc
     1081 catcaacggt gtggactttg acatggtggg tcagggcacc ggcaatccca atgatggtta
     1141 tgaggagtta aacctgaagt ccaccaaggg tgacctccag ttctccccct ggattctggt
     1201 ccctcatatc gggtatggct tccatcagta cctgccctac cctgacggga tgtcgccttt
     1261 ccaggccgcg atggtagatg gctccggata ccaagtccat cgcacaatgc agtttgaaga
     1321 tggtgcctcc cttactgtaa actaccgcta cacctacgag ggaagccaca tcaaaggaga
     1381 ggcccaggtg aaggggactg gtttccctgc tgacggtcct gtgatgacca actcgctgac
     1441 cgctgcggac tggtgcaggt cgaagaagac ttaccccaac gacaaaacca tcatcagtac
     1501 ctttaagtgg agttacacca ctggaaatgg caagcgctac cggagcactg cgcggaccac
     1561 ctacaccttt gccaagccaa tggcggctaa ctatctgaag aaccagccga tgtacgtgtt
     1621 ccgtaagacg gagctgaagc actccaagac cgagctgaac ttcaaggagt ggcaaaaggc
     1681 ctttaccgat gtgatgggca tggacgagct gtacaagggt ggcggtgggt caggtggcgg
     1741 cggaacgcgt accggtggag gcgggggttc tggaaagtct tacccaactg tgagtgctga
     1801 ttaccaggac gccgttgaga aggcgaagaa gaagctcaga ggcttcatcg ctgagaagag
     1861 atgcgctcct ctaatgctcc gtttggcatt ccactctgct ggaacctttg acaagggcac
     1921 gaagactggt ggacccttcg gaaccatcaa gcaccctgcc gaactggctc acagcgctaa
     1981 caacggtctt gacatcgctg ttaggctttt ggagccactc aaggcggagt tccctatttt
     2041 gagctacgcc gatttctacc agttggctgg cgttgttgcc gttgaggtca cgggtggacc
     2101 taaggttcca ttccaccctg gaagagagga caagcctgag ccaccaccag agggtcgctt
     2161 gcctgatccc actaagggtt ctgaccattt gagagatgtg tttggcaaag ctatggggct
     2221 tactgaccaa gatattgttg ctctatctgg gggtcacact attggagctg cacacaagga
     2281 gcgttctgga tttgagggtc cctggacctc taatcctctt attttcgaca actcatactt
     2341 cacggagttg ttgagtggtg agaaggaagg tctccttcag ctaccttctg acaaggctct
     2401 tttgtctgac cctgtattcc gccctctcgt tgataaatat gcagcggacg aagatgcctt
     2461 ctttgctgat tacgctgagg ctcaccaaaa ggcttccgag cttgggtttg ctgatgccgg
     2521 agggggtggc tctggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgtaact
     2581 cgagttaatg ttaactaact gactagcgcc cctctccctc cccctaacgt tactggccga
     2641 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     2701 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     2761 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     2821 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2881 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     2941 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     3001 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     3061 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     3121 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atggccaagc
     3181 cactgtccca agaagagtca actctgatcg agagggccac tgcaaccatt aatagcattc
     3241 ccatctctga agactatagc gtagctagtg ccgcactcag ctctgatgga cgcatattca
     3301 ccggcgttaa tgtctaccac ttcaccggcg gaccctgcgc cgaactggtc gtgctgggga
     3361 ccgcagccgc cgcggctgcc gggaatttga cgtgcattgt tgcaataggc aacgagaata
     3421 ggggcatcct gtcaccttgc ggccggtgtc ggcaagtgct gctggacctg caccccggca
     3481 tcaaggccat agtcaaggat agtgatggcc agccgaccgc cgttgggatt cgagaacttc
     3541 tgccttctgg gtacgtctgg gaaggctagg ctaagctcgc tcggctgggt cgcgggaggg
     3601 ccggtcgcag tgattcaggc ccttctggat tgtgttggtc cccagggcac gattgtcatg
     3661 cccacgcact cgggtgatct gactgatccc gcagattgga gatcgccgcc cgtgcctgcc
     3721 gattgggtgc agatccatgt cgacctgcag ccaagcttat cgataaaata aaagatttta
     3781 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3841 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     3901 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     3961 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     4021 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4081 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4141 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     4201 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4261 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     4321 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     4381 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     4441 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     4501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6601 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6661 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6721 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6781 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6841 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6901 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     6961 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     7021 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     7081 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     7141 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     7201 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     7261 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     7321 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     7381 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     7441 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     7501 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     7561 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     7621 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     7681 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     7741 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     7801 actgcgtcgc ccgggtaccc gtattc
//
