LOCUS       sequence-261079-        6700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EFF-1 A full length protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  AUTHORS   Valansi C, Moi D, Leikina E, Matveev E, Grana M, Chernomordik LV, 
            Romero H, Aguilar PS, Podbilewicz B
  TITLE     Arabidopsis HAP2/GCS1 is a gamete fusion protein homologous to 
            somatic and viral fusogens.
  JOURNAL   J Cell Biol. 2017 Mar 6;216(3):571-581. doi: 10.1083/jcb.201610093. 
            Epub 2017 Jan 30.
  PUBMED    28137780
REFERENCE   2  (bases 1 to 6700)
  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: 
            "10.1083/jcb.201610093"; journalName: "J Cell Biol"; date: 
            "2017-03-6- 6"; volume: "216"; issue: "3"; pages: "571-581"
COMMENT     Sequence Label: EFF-1::V5
FEATURES             Location/Qualifiers
     source          1..6700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..82
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2334..2375
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2466..2507
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2517..2534
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2557..2574)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2563..2787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2833..3261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2920..2939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3130..3151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3270..3290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3275..3604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3455..3590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3517..3536
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3652..3699
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3718..4092
                     /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    4222..4343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4259..4278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4313..4332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4392..4408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4392..4408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4405..4427)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4416..4432
                     /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(4440..4446,4447..4464,4465..4470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4485..4506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4623..4640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4794..5382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4874..4893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5553..6344,6345..6413))
                     /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     6176..6195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6414..6518)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggagtactgt cctccgagtc gactctagag ggtatataat ggatctcgag atatcggagc
       61 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      121 aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcgcattcc
      181 ccgtgttaat taacaggtaa gtgtcttcct cctgtttcct tcccctgcta ttctgctcaa
      241 ccttcctatc agaaactgca gtatctgtat ttttgctagc agtaatacta acggttcttt
      301 ttttctcttc acaggccacc aagcttggta ccactatgga accgccgttt gagtggtctc
      361 cccagtttat tctgcttctc ctagcagtta caacctacgg atttccactc gaggagaaat
      421 tcgatgggct cttccgtgca gagccaccac actgttccaa gacaccaatt gtccgtgctc
      481 aaacatctca aaatgcgatg tcatcaattg caagaggaat gcaaatgcaa ttctcaattg
      541 gtttgcacac tgcagtctgc ttcagattat atgaagacac tcaactcgca tcccaagaga
      601 tcaatgatga tgaaaatgct ggaaatcaga catcacttct tcatactatt cgattggaaa
      661 agcttgaaca tcaccatccg ataactcagc ggtacacttt tggaatccct gaagttcatg
      721 ccagttgtat ctgtgagtgt gatgccacat cttcaacatg taccgcagaa tctcatcagt
      781 ttaccgcttg tccagagtct gataagtctg atgaaacatc ttcatgctac cgcacgttct
      841 tcccaaatca gacaccaatc gggtgcagtg aagacgacat ccccaagctt tgctgtgacg
      901 ttcgattcaa gccatataaa aatatgacat tcctcgcagt caaactagaa caacctacaa
      961 cgtatgcaac atttgtctac gcagcttatg attttgtaaa tggatattgg gtggaaaaag
     1021 ataaaactaa aatccgttct caattagatg gtggaacaca ggataggcat ctcgaccaaa
     1081 aacgtcgaat ttcattggct gtaactgccg gaggacgagc atctcatcaa ctggagacag
     1141 gaatgtattt ctcaagaaca agtaatggag gagaaacaga agaactgaga atgcaaccat
     1201 tgaatgagat tacagacaat aactttgacc gtcttggatg gtacagaatg gacgattctg
     1261 gtcattttca tgtgaataat ggagttgtga aaatggatga tattcacaag gctaaagtga
     1321 aaaattgcaa agagcaaact tataaatcaa ttctctctgc aaatcattat atgccaggac
     1381 atttcaattt gactcgtcca cttgaagtta tcaaaccgtg gattcaatct gctcgaattt
     1441 ttgatagttc tctcagacaa gctgttgtaa cacatgccga aggaactaat cttcaaattt
     1501 caattcactt ggatgacgag gtggaaagtc aaaaccttgt cttcttccat aatgcatcaa
     1561 gaattcgtga cttcagtgga tccatcatcg ttgattcaaa atcaaatcga ttgttcaact
     1621 tgacagtata tgaggcttct ggaaaaattg atggatcagt gaagatgtca actggatttg
     1681 gatctgatac aattcacaca ttcactgctt atgttagtga tcttcatgct tcaaaccgtt
     1741 ctatgattat tccactgcca gccatcgttg gacaaggagc cagagcaatc tgtctccgag
     1801 cagattcaat ggctgatatt gacaaaattt gtcatgtcat tgagtatttc gagagccctt
     1861 tggaaatcga tcttgttgag ggaaaatggc acgaaatgat tggaacctgc ccaacttgta
     1921 atcaaatcaa tttcaatgga atgatgaaat ttctgaatcc agctcattgg atcaaaggaa
     1981 tcagttcgat tggtgacggc gttatgattg ctacggatat tgttgtgtat ctcggagttt
     2041 tgtgtattct gtatcttctt attacgaaga ttattgttcc gttggttcgt tgctgggtat
     2101 gccccatgtc gattttctgc aatgggtcct cttcgtcgtc caaaaacaaa aatgataaga
     2161 gaaggaagga gagagaggag aggcgcagaa aggacaaatt tgtctcagaa tccgaagatg
     2221 gtgcaagaag tagttccgag ccccacgaca ctttggcgag ataccatgga aatcactcgg
     2281 agcggcacta tagcagtagc cagtacattg gtctagaggg cccgcggttc gaaggtaagc
     2341 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttgagct agtccagtgt
     2401 ggtggaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgcggt
     2461 tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc
     2521 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2581 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2641 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2701 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2761 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2821 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2881 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2941 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3001 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3061 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3121 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3181 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3241 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3301 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3361 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3421 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3481 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3541 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3601 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     3661 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     3721 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     3781 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     3841 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     3901 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     3961 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     4021 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     4081 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4141 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4201 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4321 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4381 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4441 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4501 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4561 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4621 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4681 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5521 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5581 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5641 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5701 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5761 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5821 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5881 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5941 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6001 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6061 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6121 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6181 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6241 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6301 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6361 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6421 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6481 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6541 cctgacgtcg acggatcggg agatcgtacc gagctcttac gcgggtcgaa gcggagtact
     6601 gtcctccgag tggagtactg tcctccgagc ggagtactgt cctccgagtc gagggtcgaa
     6661 gcggagtact gtcctccgag tggagtactg tcctccgagc
//
