LOCUS       sequence-139746-        6957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-dL5(E52D)-mCer3 fusion protein in peroxisomes of 
            mammalian cells with a C-terminal PTS (SKL). (MBIC5, dL5**, FAP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6957)
  AUTHORS   Telmer CA, Verma R, Teng H, Andreko S, Law L, Bruchez MP
  TITLE     Rapid, specific, no-wash, far-red fluorogen activation in 
            subcellular compartments by targeted fluorogen activating proteins.
  JOURNAL   ACS Chem Biol. 2015 May 15;10(5):1239-46. doi: 10.1021/cb500957k. 
            Epub 2015 Feb 16.
  PUBMED    25650487
REFERENCE   2  (bases 1 to 6957)
  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: "10.1021/cb500957k";
            journalName: "ACS Chem Biol"; date: "2015-05-15- 15"; volume: "10"; 
            issue: "5"; pages: "1239-46"
COMMENT     Sequence Label: pcDNA3.1-KozATG-myc-dL5-2XG4S-mCer3-SKL
FEATURES             Location/Qualifiers
     source          1..6957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      942..951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             951..980
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             996..1715
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     CDS             join(1773..1775,1776..1778,1779..2489)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1818..1839)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2079..2098)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2426..2447
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2551..2568)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2557..2781
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2827..3255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2914..2933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3124..3145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3264..3284)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3269..3598
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3449..3584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3511..3530
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3665..4459
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3719..3738)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4329..4348
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4633..4754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4670..4689)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4724..4743
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4803..4819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4803..4819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4816..4838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4827..4843
                     /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(4851..4857,4858..4875,4876..4881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4896..4917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5034..5051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5205..5790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5285..5304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5961..6752,6753..6821))
                     /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     6584..6603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6822..6926)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
      901 tagcgtttaa acttaagctt gccaccggat cccttgaatt cgccaccatg gagcagaagc
      961 tgatcagcga ggaggacctg ggcctttctg gcctgcaggc cgtcgttacc caagaaccta
     1021 gtgttaccgt tagcccaggt ggtactgtta tacttacttg tggaagtggt acgggtgccg
     1081 tcacatctgg tcattatgca aattggtttc aacaaaaacc aggacaagct ccaagagctt
     1141 tgatttttga tactgataag aagtattctt ggaccccagg tagattttct ggatctttgc
     1201 tgggagcaaa ggcagctttg acaatatcag atgctcagcc tgaggacgaa gccgagtatt
     1261 actgttctct tagcgacgtg gatggctact tgtttggcgg tggaacacaa ctgacggttc
     1321 tgtccggtgg tggcggctct ggtggcggtg gcagcggcgg tggtggttcc ggaggcggcg
     1381 gttctcaggc tgtggtgact caggagccgt cagtgactgt gtccccagga gggacagtca
     1441 ttctcacttg tggctccggc actggagctg tcaccagtgg tcattatgcc aactggttcc
     1501 agcagaagcc tggccaagcc cccagggcac ttatatttga caccgacaag aagtattcct
     1561 ggacccctgg ccgattctca ggctccctcc ttggggccaa ggctgccctg accatctcgg
     1621 atgcgcagcc tgaagatgag gctgagtatt actgttcgct ctccgacgtt gacggttatc
     1681 tgttcggagg aggcacccag ctgaccgtcc tctccgcggc cggtctggcc catatgggag
     1741 gcggtgggtc tggtggcggt ggcagcaccg gtatggtgag caagggcgag gagctgttca
     1801 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1861 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1921 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc tggggcgtgc
     1981 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2041 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2101 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2161 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacgcc atccacggca
     2221 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc ggcctcaact
     2281 gcaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2341 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     2401 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2461 tcactctcgg catggacgag ctgtacaaga gcaagctgta actcgaggca gctgtcgact
     2521 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2581 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2641 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2701 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2761 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2821 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2881 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2941 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3001 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3061 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3121 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3181 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3241 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3361 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3421 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3481 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3541 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3601 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3661 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3721 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3781 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3841 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3901 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3961 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4021 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4081 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4141 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4201 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4261 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4321 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4381 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4441 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4501 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4561 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4621 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4681 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4741 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4801 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4861 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4921 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4981 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5041 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5101 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5161 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5221 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5281 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5341 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5401 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5461 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5521 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5581 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5641 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5701 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     5761 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6061 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6841 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6901 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
