LOCUS       sequence-261087-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EFF-1 fused to mCherry fluorescent protein cloned into the pGene 
            vector for inducible expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  AUTHORS   Perez-Vargas J, Krey T, Valansi C, Avinoam O, Haouz A, Jamin M, 
            Raveh-Barak H, Podbilewicz B, Rey FA
  TITLE     Structural basis of eukaryotic cell-cell fusion.
  JOURNAL   Cell. 2014 Apr 10;157(2):407-419. doi: 10.1016/j.cell.2014.02.020.
  PUBMED    24725407
REFERENCE   2  (bases 1 to 7424)
  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: 
            "10.1016/j.cell.2014.02"; journalName: "Cell"; date: "2014-04-10- 
            10"; volume: "157"; issue: "2"; pages: "407-419"
COMMENT     Sequence Label: pGENE:EFF-1::mCherry::V5
FEATURES             Location/Qualifiers
     source          1..7424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..33
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2285..2326
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2336..3043
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2481..2499)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2749..2768
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3141..3182
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3192..3209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3232..3249)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3238..3462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3508..3936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3595..3614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3805..3826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3945..3965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3950..4279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4130..4265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4192..4211
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4327..4374
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4393..4767
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4897..5018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4934..4953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4988..5007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5067..5083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5067..5083)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5080..5102)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5091..5107
                     /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(5115..5121,5122..5139,5140..5145))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5160..5181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5298..5315)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5469..6057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5549..5568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6228..7019,7020..7088))
                     /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     6851..6870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7089..7193)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatatcggag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
       61 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      121 acgcgcattc cccgtgttaa ttaacaggta agtgtcttcc tcctgtttcc ttcccctgct
      181 attctgctca accttcctat cagaaactgc agtatctgta tttttgctag cagtaatact
      241 aacggttctt tttttctctt cacaggccac caagcttggt accactatgg aaccgccgtt
      301 tgagtggtct ccccagttta ttctgcttct cctagcagtt acaacctacg gatttccact
      361 cgaggagaaa ttcgatgggc tcttccgtgc agagccacca cactgttcca agacaccaat
      421 tgtccgtgct caaacatctc aaaatgcgat gtcatcaatt gcaagaggaa tgcaaatgca
      481 attctcaatt ggtttgcaca ctgcagtctg cttcagatta tatgaagaca ctcaactcgc
      541 atcccaagag atcaatgatg atgaaaatgc tggaaatcag acatcacttc ttcatactat
      601 tcgattggaa aagcttgaac atcaccatcc gataactcag cggtacactt ttggaatccc
      661 tgaagttcat gccagttgta tctgtgagtg tgatgccaca tcttcaacat gtaccgcaga
      721 atctcatcag tttaccgctt gtccagagtc tgataagtct gatgaaacat cttcatgcta
      781 ccgcacgttc ttcccaaatc agacaccaat cgggtgcagt gaagacgaca tccccaagct
      841 ttgctgtgac gttcgattca agccatataa aaatatgaca ttcctcgcag tcaaactaga
      901 acaacctaca acgtatgcaa catttgtcta cgcagcttat gattttgtaa atggatattg
      961 ggtggaaaaa gataaaacta aaatccgttc tcaattagat ggtggaacac aggataggca
     1021 tctcgaccaa aaacgtcgaa tttcattggc tgtaactgcc ggaggacgag catctcatca
     1081 actggagaca ggaatgtatt tctcaagaac aagtaatgga ggagaaacag aagaactgag
     1141 aatgcaacca ttgaatgaga ttacagacaa taactttgac cgtcttggat ggtacagaat
     1201 ggacgattct ggtcattttc atgtgaataa tggagttgtg aaaatggatg atattcacaa
     1261 ggctaaagtg aaaaattgca aagagcaaac ttataaatca attctctctg caaatcatta
     1321 tatgccagga catttcaatt tgactcgtcc acttgaagtt atcaaaccgt ggattcaatc
     1381 tgctcgaatt tttgatagtt ctctcagaca agctgttgta acacatgccg aaggaactaa
     1441 tcttcaaatt tcaattcact tggatgacga ggtggaaagt caaaaccttg tcttcttcca
     1501 taatgcatca agaattcgtg acttcagtgg atccatcatc gttgattcaa aatcaaatcg
     1561 attgttcaac ttgacagtat atgaggcttc tggaaaaatt gatggatcag tgaagatgtc
     1621 aactggattt ggatctgata caattcacac attcactgct tatgttagtg atcttcatgc
     1681 ttcaaaccgt tctatgatta ttccactgcc agccatcgtt ggacaaggag ccagagcaat
     1741 ctgtctccga gcagattcaa tggctgatat tgacaaaatt tgtcatgtca ttgagtattt
     1801 cgagagccct ttggaaatcg atcttgttga gggaaaatgg cacgaaatga ttggaacctg
     1861 cccaacttgt aatcaaatca atttcaatgg aatgatgaaa tttctgaatc cagctcattg
     1921 gatcaaagga atcagttcga ttggtgacgg cgttatgatt gctacggata ttgttgtgta
     1981 tctcggagtt ttgtgtattc tgtatcttct tattacgaag attattgttc cgttggttcg
     2041 ttgctgggta tgccccatgt cgattttctg caatgggtcc tcttcgtcgt ccaaaaacaa
     2101 aaatgataag agaaggaagg agagagagga gaggcgcaga aaggacaaat ttgtctcaga
     2161 atccgaagat ggtgcaagaa gtagttccga gccccacgac actttggcga gataccatgg
     2221 aaatcactcg gagcggcact atagcagtag ccagtacatt ggtctagagg gcccgcggtt
     2281 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtatggt
     2341 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     2401 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2461 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     2521 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     2581 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2641 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     2701 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     2761 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     2821 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     2881 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     2941 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     3001 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagaccggta attaccggtt
     3061 gagctagtcc agtgtggtgg aattctgcag atatccagca cagtggcggc cgctcgagtc
     3121 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     3181 cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     3241 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3301 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3361 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3421 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3481 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3721 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3841 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3901 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4201 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4261 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4321 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     4381 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     4441 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     4501 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     4561 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     4621 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     4681 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     4741 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     4801 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4861 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4981 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5041 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5101 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5161 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5221 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5281 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5341 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5401 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5461 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5521 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5581 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5641 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5701 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5761 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5821 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5881 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5941 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6001 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6061 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6121 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6181 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6241 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6301 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6361 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6421 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6481 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6541 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6601 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6661 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6721 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6781 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6841 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6901 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6961 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7021 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7081 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7141 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7201 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gtaccgagct cttacgcggg
     7261 tcgaagcgga gtactgtcct ccgagtggag tactgtcctc cgagcggagt actgtcctcc
     7321 gagtcgaggg tcgaagcgga gtactgtcct ccgagtggag tactgtcctc cgagcggagt
     7381 actgtcctcc gagtcgactc tagagggtat ataatggatc tcga
//
