LOCUS       sequence-481500-        8230 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Cell fusion assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8230)
  AUTHORS   Moi D, Nishio S, Li X, Valansi C, Langleib M, Brukman NG, Flyak K, 
            Dessimoz C, de Sanctis D, Tunyasuvunakool K, Jumper J, Grana M, 
            Romero H, Aguilar PS, Jovine L, Podbilewicz B
  TITLE     Discovery of archaeal fusexins homologous to eukaryotic HAP2/GCS1 
            gamete fusion proteins.
  JOURNAL   Nat Commun. 2022 Jul 6;13(1):3880. doi: 10.1038/s41467-022-31564-1.
  PUBMED    35794124
REFERENCE   2  (bases 1 to 8230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-07-6"; volume: "13(1):3880. doi"; pages: " 
            10.1038/s41467-022-31564-1"
COMMENT     Sequence Label: pCI_Fsx1-V5_GFPnes
FEATURES             Location/Qualifiers
     source          1..8230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3714..3755
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3765..3782
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    join(3799..4373,4374..4376,4377..4385)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3966..3983)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4193..4212
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4386..4388,4389..4391,4392..5099)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(4386..4388,4389..4391,4392..5099)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(4431..4452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4692..4711)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5039..5060
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5222..5241)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5287..5342
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5341..5360)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5733..5749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5733..5749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5746..5768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5757..5773
                     /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(5781..5787,5788..5805,5806..5811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5826..5847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5906..6101
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5952..6087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6107..6241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6144..6163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6198..6217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6309..6326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6480..7068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6560..6579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7239..8030,8031..8099))
                     /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     7862..7881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8100..8204)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccmggsggg gsggggsggg gggrgggggg
      541 ggggggggng nggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgagag
     1741 ttcgaagcct taagtacgcg taggcgcgcc ttaattaacg tttaaacgct agcggtacca
     1801 tgagacgtgc agcattgatt ctggctttcg tactgttcat cggtctttcc tcggctactg
     1861 ttacctcggc tgattcaatc acgtataact ctggaaccag cgagttcttc gacggagatg
     1921 tctttgcaat agaggtaacc gctgaccagt cgactgatga gatagatatc tacctcggtg
     1981 cttctgagct ttcggaaaag acggacggcg aggtcaacca ggacctcagt attgagttca
     2041 cgcaccagga ttccaagctg aagtattcta cttcgacttc tgacgagcta cgggacattg
     2101 tcactcttac aacttactac gaggacgggt ttgacacgga gcaggatgct atagacgcca
     2161 tcaaatcaga ctgttacgat ctgaatcaga acgggaatgg aagcggaagg tacagcaggt
     2221 actactcagt tacatctcct gtctatgatt acgagatcta ttgtttccag aagaacgaga
     2281 aactggctac tccagcatac atagacaacc cggatgagat cttcactgca aaggctgagc
     2341 tacaggccgg tgacaagacg attcagtcag ctacattgtc taacggtgac gccggagacg
     2401 gaacagtcac tgacctcggt gactcaaaga ttagctggaa cgggaacctg gatttaggag
     2461 cttctgaacc ggagaatagc cgagtcattg ctttgtactc gaatgatttt gagaacgggt
     2521 ggaggatcgg caataagcag tcgtacgagg actacaagac gttcatcggt ggcggggatg
     2581 cctacgatct cctgattgac tggcaggacg ggacttacac cgcctctgaa gtagaggatg
     2641 agctggtgaa tactgatgcg aaccaggctg ttgaggaggc cagttctagc acgacagact
     2701 tggtgaacgc gaaggttaag gactcgtctc ttgacaccgg ttcgttcgtg tatgataccc
     2761 ctgagctgct ttcctacccg tccttcacag tctacgtcga tgccggagaa aacggataca
     2821 tcgaggtaac gaagccgact ggtgaccccg atatcatctc cacttcatct acagaaatta
     2881 aggagggaga tgagggaact gtaaccgcca ccgttgagaa tgtcggagat ggtgagggcg
     2941 agttctctgg aaggctttcc agctgcggtg aaggcttcag tattgtcgac gaccagaata
     3001 cgaagaatgt gggtgccgga gaatcggtga catacagttt tgatgtcgct ttcagttctg
     3061 tcagcagtga gtcgaaggag atctcgggca gctgcacgtt cgaggtgaac ggcgtcgagt
     3121 ccagtgattc caccagtgtt tctgtgaccg gtatccagca gagtgagtgt aatccaggcg
     3181 atcagagacg ggaaaagaat gagaatgacc gctgggagat ctacacctgc caggacaacg
     3241 gcctgaccta cgaatatgat gtaacttgtg cagaggacga gaaggctgta gctcagggag
     3301 ataaccagtt ttcctgtgag aaacaggacg atgattcagg tggcggagac aacactggat
     3361 ctgactccgg actgttctcg aatctgttcg gaggttctgg ttccggagat ctgcttacac
     3421 aggttcacac agctctgtct atcctcgcgg ggcttgttgc aggcttcttc ggttatagag
     3481 gagcccgatg gattcacggc gagactgata taaaaggagg cttcaagctg gagtcccgga
     3541 acgtaagccg tgttaagaga ggtagtcctg tagccggtat tgtcggagct gtactcgggt
     3601 ttgtcgtagg atacggtgtt gcctcagtct tccatccagt agttcagatt atagtggttc
     3661 tgggaattgc tgttggcttg tactatttca ggctagaggg cccgcggttc gaaggtaagc
     3721 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     3781 attagcccgg gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     3841 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     3901 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     3961 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     4021 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     4081 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     4141 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     4201 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     4261 tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     4321 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     4381 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4441 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     4501 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     4561 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     4621 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     4681 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     4741 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     4801 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     4861 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     4921 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     4981 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5041 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5101 tgagcggcgg cctgcagaag aagctggagg agctggagct ggactaatgt acaagtaaag
     5161 cggccggccg ccagcacagt ggtcgatcga ccaatgccct ggctcacaaa taccactgag
     5221 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     5281 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     5341 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     5401 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     5461 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     5521 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     5581 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     5641 tagctgtccc tcttctctta tggagatccc tcgacctgca ccgtcgacca gctggtcgac
     5701 ggtgcaccgt cgaccagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5821 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5881 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     5941 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6001 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6061 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     6121 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6181 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     6241 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7201 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7261 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7321 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7381 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7441 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7501 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7561 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7621 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7681 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7741 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7801 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7861 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7921 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7981 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8041 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8221 gtgccacctg
//
