LOCUS       sequence-322850-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Detect the long Peroxisomes-mitochondria contact .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  AUTHORS   Vallese F, Catoni C, Cieri D, Barazzuol L, Ramirez O, Calore V, 
            Bonora M, Giamogante F, Pinton P, Brini M, Cali T
  TITLE     An expanded palette of improved SPLICS reporters detects multiple 
            organelle contacts in vitro and in vivo.
  JOURNAL   Nat Commun. 2020 Nov 27;11(1):6069. doi: 10.1038/s41467-020-19892-6.
  PUBMED    33247103
REFERENCE   2  (bases 1 to 6746)
  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 
            Commun."; date: "2020-11-27"; volume: "11(1):6069. doi"; pages: " 
            10.1038/s41467-020-19892-6"
COMMENT     Sequence Label: SPLICS Po-Mt Long P2A
FEATURES             Location/Qualifiers
     source          1..6746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..76)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        248..627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     782..802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     876..895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        876..894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             933..980
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1497..1553
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2350..2367)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2356..2580
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2626..3054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2713..2732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2923..2944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3063..3083)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3068..3397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3248..3383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3310..3329
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3464..4258
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPAICPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3518..3537)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4128..4147
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4432..4553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4469..4488)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4523..4542
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4602..4618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4602..4618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4615..4637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4626..4642
                     /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(4650..4656,4657..4674,4675..4680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4695..4716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4833..4850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5004..5592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5084..5103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5763..6554,6555..6623))
                     /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     6386..6405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6624..6728)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
       61 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
      121 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      181 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      241 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      301 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      361 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      421 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      481 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      541 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      601 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      661 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      721 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      781 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      841 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      901 caagctggct agcgtttaaa cttaagctta tgcgggacca catggtgctg cacgagtacg
      961 tgaacgccgc cggcatcacc ggaggcgatg gaggctccgg aggaggctct aagctgaaca
     1021 attctggcgc ccctcacagg gagaagaggg gaggagagac cgacgagttc tccaatgtgc
     1081 ggagaggccg gggccacaga atgcagcacc tgtctgaggg cacaaaggga aggcaagtgg
     1141 gctctggagg cgacggagag agatggggca gcgatagggg ctctcgcggc agcctgaacg
     1201 agcagatcgc cctggtgctg atgaggctgc aggaggatat gcagaatgtg ctgcagcgcc
     1261 tgcagaagct ggagaccctg acagccctgc aggccaagag ctccaccagc acactgcaga
     1321 ccgcaccaca gcctacaagc cagaggccat cctggtggcc ctttgagatg tcccctggcg
     1381 tgctgacatt cgccatcatc tggcccttca tcgcccagtg gctggtgtat ctgtactatc
     1441 agaggcgccg gagaaagctg aacggatccc tgaaggagct ggcatgcggc agcggagcaa
     1501 ccaacttctc cctgctgaag caggcaggcg acgtggagga gaatcccggc cctcggtctg
     1561 aattcatggt gggccgcaat tccgcaatcg cagcaggagt gtgcggcgcc ctgttcatcg
     1621 gctactgtat ctattttgac cggaagcgga gaagcgatcc aaattccaag ggcgaggagc
     1681 tgttcacagg cgtggtgccc atcctggtgg agctggacgg cgatgtgaac ggccacaagt
     1741 tttctgtgcg gggagaggga gagggcgacg caaccatcgg caagctgaca ctgaagttca
     1801 tctgcaccac aggcaagctg cccgtgcctt ggccaaccct ggtgaccaca ctgacatacg
     1861 gcgtgcagtg ttttagcagg tatcctgacc acatgaagcg ccacgatttc tttaagtccg
     1921 ccatgccaga gggctacgtg caggagcgga ccatctcttt caaggacgat ggcaagtata
     1981 agaccagagc cgtggtgaag tttgagggcg acacactggt gaacaggatc gagctgaagg
     2041 gcaccgactt caaggaggat ggcaatatcc tgggccacaa gctggagtac aactttaata
     2101 gccacaacgt gtatatcaca gccgataagc agaagaacgg catcaaggcc aatttcaccg
     2161 tgagacacaa tgtggaggac ggctccgtgc agctggccga tcactaccag cagaacacac
     2221 caatcggcga cggccctgtg ctgctgcctg ataatcacta tctgtctacc cagacagtgc
     2281 tgagcaagga ccccaacgag aagggcacct gactcgagtc tagagggccc gtttaaaccc
     2341 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2401 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2461 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2521 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2581 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     2641 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2701 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2761 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2821 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2881 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2941 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3001 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3061 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3121 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3241 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3301 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3361 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3421 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3481 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3541 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3601 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     3661 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3721 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3781 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3841 tgatccggct atctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3901 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3961 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4021 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4081 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4141 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4201 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4261 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4321 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4381 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4441 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4501 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4561 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4621 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4681 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4741 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6601 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6661 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6721 gttccgcgca catttccccg aaaagt
//
