LOCUS       sequence-322840-        6557 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 6557)
  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 6557)
  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 Short P2A
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(136..156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        141..470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      321..456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     383..402
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             537..1331
                     /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(591..610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1201..1220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1505..1626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1542..1561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1596..1615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1675..1691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1675..1691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1688..1710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1699..1715
                     /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(1723..1729,1730..1747,1748..1753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1768..1789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1906..1923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2077..2665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2157..2176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2836..3627,3628..3696))
                     /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     3459..3478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3697..3801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3876..3895)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4067..4446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4447..4650
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4601..4621
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4695..4714
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4695..4713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4752..4799
                     /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             4980..5036
                     /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(5980..5997)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5986..6210
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6256..6557,1..127)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6343..6362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6553..6557,1..17)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
       61 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      121 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
      181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
      241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
      421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
      481 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
      541 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      601 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      661 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
      721 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
      781 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
      841 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
      901 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
      961 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     1021 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     1081 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1141 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     1201 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     1261 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     1321 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     1381 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1501 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1621 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     1681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     3841 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3901 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     3961 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     4021 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
     4081 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4141 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     4201 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4261 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4321 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4381 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4441 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     4501 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4561 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     4621 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
     4681 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gcgtttaaac
     4741 ttaagcttat gcgggaccac atggtgctgc acgagtacgt gaacgccgcc ggcatcaccg
     4801 gaggcgatgg aggcagcgga ggaggctcca agctgtggtg gcccttcgag atgtctcctg
     4861 gcgtgctgac attcgccatc atctggccct ttatcgccca gtggctggtg tatctgtact
     4921 atcagcggag aaggcgcaag ctgaatggat ccctgaagga gctggcatgc ggcagcggag
     4981 caaccaactt ctccctgctg aagcaggcag gcgacgtgga ggagaatccc ggccctcggt
     5041 ctgaattcat gagcaaggga gaggagctgt tcaccggagt ggtgcctatc ctggtggagc
     5101 tggacggcga tgtgaatggc cacaagtttt ccgtgcgcgg agagggagag ggcgacgcaa
     5161 caatcggcaa gctgaccctg aagttcatct gcaccacagg caagctgccc gtgccttggc
     5221 caaccctggt gaccacactg ggctacggcc tgcagtgttt tgcccggtat ccagaccaca
     5281 tgaagagaca cgatttcttt aagtccgcca tgcccgaggg ctacgtgcag gagaggacaa
     5341 tctctttcaa ggacgatggc aagtataaga cacgcgccgt ggtgaagttt gagggcgaca
     5401 ccctggtgaa ccggatcgag ctgaagggca cagacttcaa ggaggatggc aatatcctgg
     5461 gccacaagct ggagtacaac tttaattccc acaacgtgta tatcaccgcc gataagcaga
     5521 agaacggcat caaggccaat ttcacagtgc ggcacaatgt ggaggacggc tctgtgcagc
     5581 tggccgatca ctaccagcag aacaccccta tcggcgacgg ccctgtgctg ctgcctgata
     5641 atcactacct gtcttatcag accgtgctga gcaaggaccc aaacgagaag ggcacaggag
     5701 gcgatggagg cagcggagga ggctccaagc tgagagtgtt cctggccctg cccatcatca
     5761 tggtggtggc cttttctatg tgcatcatct gtctgctgat ggcaggcgac acctggacag
     5821 agaccctggc ctacgtgctg ttttggggcg tggccagcat cggcacattc tttatcatcc
     5881 tgtataacgg caaggacttc gtggatgccc ccaggctggt gcagaaggag aagatcgatt
     5941 gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     6001 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     6061 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     6121 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     6181 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     6241 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6301 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6361 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6421 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6481 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6541 cgttctttaa tagtgga
//
