LOCUS       sequence-322839-        6893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Detect the long Peroxisomes-Endoplasmic reticulum contact .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6893)
  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 6893)
  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-ER Long P2A
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             921..968
                     /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             1485..1541
                     /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(2485..2502)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2491..2715
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2761..3189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2848..2867)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3058..3079
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3198..3218)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3203..3532
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3383..3518
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3445..3464
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3599..4393
                     /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(3653..3672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4263..4282
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4567..4688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4604..4623)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4658..4677
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4737..4753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4737..4753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4750..4772)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4761..4777
                     /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(4785..4791,4792..4809,4810..4815))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4830..4851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4968..4985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5139..5727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5219..5238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5898..6689,6690..6758))
                     /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     6521..6540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6759..6863)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact taagcttatg cgggaccaca tggtgctgca cgagtacgtg aacgccgccg
      961 gcatcaccgg aggcgatgga ggctccggag gaggctctaa gctgaacaat tctggcgccc
     1021 ctcacaggga gaagagggga ggagagaccg acgagttctc caatgtgcgg agaggccggg
     1081 gccacagaat gcagcacctg tctgagggca caaagggaag gcaagtgggc tctggaggcg
     1141 acggagagag atggggcagc gataggggct ctcgcggcag cctgaacgag cagatcgccc
     1201 tggtgctgat gaggctgcag gaggatatgc agaatgtgct gcagcgcctg cagaagctgg
     1261 agaccctgac agccctgcag gccaagagct ccaccagcac actgcagacc gcaccacagc
     1321 ctacaagcca gaggccatcc tggtggccct ttgagatgtc ccctggcgtg ctgacattcg
     1381 ccatcatctg gcccttcatc gcccagtggc tggtgtatct gtactatcag aggcgccgga
     1441 gaaagctgaa cggatccctg aaggagctgg catgcggcag cggagcaacc aacttctccc
     1501 tgctgaagca ggcaggcgac gtggaggaga atcccggccc tcggtctgaa ttcatgagca
     1561 agggagagga gctgttcacc ggagtggtgc ctatcctggt ggagctggac ggcgatgtga
     1621 atggccacaa gttttccgtg cgcggagagg gagagggcga cgcaacaatc ggcaagctga
     1681 ccctgaagtt catctgcacc acaggcaagc tgcccgtgcc ttggccaacc ctggtgacca
     1741 cactgggcta cggcctgcag tgttttgccc ggtatccaga ccacatgaag agacacgatt
     1801 tctttaagtc cgccatgccc gagggctacg tgcaggagag gacaatctct ttcaaggacg
     1861 atggcaagta taagacacgc gccgtggtga agtttgaggg cgacaccctg gtgaaccgga
     1921 tcgagctgaa gggcacagac ttcaaggagg atggcaatat cctgggccac aagctggagt
     1981 acaactttaa ttcccacaac gtgtatatca ccgccgataa gcagaagaac ggcatcaagg
     2041 ccaatttcac agtgcggcac aatgtggagg acggctctgt gcagctggcc gatcactacc
     2101 agcagaacac ccctatcggc gacggccctg tgctgctgcc tgataatcac tacctgtctt
     2161 atcagaccgt gctgagcaag gacccaaacg agaagggcac aggaggcgat ggaggcagcg
     2221 gaggaggctc caagctgaga gtgttcctgg ccctgcccat catcatggtg gtggcctttt
     2281 ctatgtgcat catctgtctg ctgatggcag gcgacacctg gacagagacc ctggcctacg
     2341 tgctgttttg gggcgtggcc agcatcggca cattctttat catcctgtat aacggcaagg
     2401 acttcgtgga tgcccccagg ctggtgcaga aggagaagat cgattgactc gagtctagag
     2461 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2521 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2581 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2641 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2701 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2761 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2821 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2881 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2941 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3001 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3061 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3121 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3181 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3301 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3361 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3421 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3481 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3541 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3601 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3661 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3721 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3781 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3841 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3901 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3961 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4021 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4081 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4141 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4201 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4261 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4321 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4381 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4441 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4501 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4561 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4681 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4801 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5941 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6001 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6061 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgt
//
