LOCUS       sequence-323257-        6165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the non-fluorescent GFP1-10 moiety on the cytosolic face 
            of the OMM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6165)
  AUTHORS   Cieri D, Vicario M, Giacomello M, Vallese F, Filadi R, Wagner T, 
            Pozzan T, Pizzo P, Scorrano L, Brini M, Cali T
  TITLE     SPLICS: a split green fluorescent protein-based contact site sensor 
            for narrow and wide heterotypic organelle juxtaposition.
  JOURNAL   Cell Death Differ. 2018 Jun;25(6):1131-1145. doi: 
            10.1038/s41418-017-0033-z. Epub 2017 Dec 11.
  PUBMED    29229997
REFERENCE   2  (bases 1 to 6165)
  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: 
            "10.1038/s41418-017-0033-z"; journalName: "Cell Death Differ"; date:
            "2018-06"; volume: "25"; issue: "6"; pages: "1131-1145"
COMMENT     Sequence Label: OMMGFP 1-10
FEATURES             Location/Qualifiers
     source          1..6165
                     /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"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1718..1736)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1719..1736)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1756..1773)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1762..1986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2032..2460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2119..2138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2329..2350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2469..2489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2474..2803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2654..2789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2716..2735
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2870..3664
                     /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(2924..2943)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3534..3553
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3838..3959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3875..3894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3929..3948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4008..4024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4008..4024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4021..4043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4032..4048
                     /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(4056..4062,4063..4080,4081..4086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4101..4122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4239..4256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4410..4998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4490..4509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5169..5960,5961..6029))
                     /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     5792..5811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6030..6134)
                     /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 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcatggtgg gccggaacag
      961 cgccatcgcc gcgggcgtgt gcggtgccct cttcataggg tactgcatct actttgaccg
     1021 caaaaggcgg agtgacccca actccaaagg agaagaactg tttaccggcg tggtgccaat
     1081 tctcgtggaa ctggatggcg atgtgaatgg ccacaaattt tctgtcagag gagagggtga
     1141 aggtgatgcc acaatcggaa agctcaccct gaaattcatc tgcaccactg gaaagctccc
     1201 tgtgccatgg ccaacactgg tcactaccct gacctacggc gtgcagtgct tttccagata
     1261 cccagaccat atgaagaggc atgacttttt caagagcgcc atgcccgagg gctatgtgca
     1321 ggagagaacc atctctttca aagatgacgg gaaatacaag acccgcgctg tggtcaagtt
     1381 cgaaggagac acactggtga atagaatcga gttgaagggc acagacttta aggaagatgg
     1441 aaacattctc ggccacaagc tggaatacaa ctttaactcc cacaatgtgt acatcacagc
     1501 cgacaagcaa aagaatggca tcaaggctaa cttcacagtc agacacaacg tcgaggatgg
     1561 aagcgtgcag ctggccgacc attatcaaca gaacactcca atcggcgacg gccctgtgct
     1621 cctcccagac aaccattacc tgtccaccca gacagtcctg agcaaagatc caaatgaaaa
     1681 aggaacataa ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta
     1741 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1801 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1861 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1921 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     1981 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2041 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2101 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2161 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2221 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2281 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2341 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2401 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2461 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2641 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2701 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2761 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2821 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2881 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2941 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3001 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3061 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3121 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3181 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3241 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3301 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3361 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3421 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3481 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3541 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3601 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3661 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3721 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3781 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3841 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3901 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3961 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4021 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4081 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4141 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4201 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4261 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4321 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4381 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4441 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4501 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4561 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4621 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4681 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4741 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4801 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4861 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4921 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4981 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5041 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5101 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5161 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5221 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5281 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5341 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5401 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5461 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5521 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5581 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5641 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5701 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5761 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5821 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5881 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5941 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6001 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6061 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6121 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
