LOCUS       sequence-481102-        8282 bp    DNA     circular SYN 29-JUL-2025
DEFINITION  Cell fusion assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8282)
  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 8282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 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_EFF-1-V5_GFPnes
FEATURES             Location/Qualifiers
     source          1..8282
                     /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             3793..3834
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    join(3851..4425,4426..4428,4429..4437)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4018..4035)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4245..4264
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4438..4440,4441..4443,4444..5151)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(4438..4440,4441..4443,4444..5151)
                     /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(4483..4504)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4744..4763)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5091..5112
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5274..5293)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5339..5394
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5393..5412)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5785..5801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5785..5801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5798..5820)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5809..5825
                     /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(5833..5839,5840..5857,5858..5863))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5878..5899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5958..6153
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6004..6139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6159..6293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6196..6215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6250..6269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6361..6378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6532..7120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6612..6631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7291..8082,8083..8151))
                     /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     7914..7933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8152..8256)
                     /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 gccrggsggg ggggggsggg gggggggggg
      541 gggnggggsg rggcggagag 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 gkgggggtgc cgggcggggc
     1261 ggggccgcck 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 agctatggaa
     1801 ccgccgtttg agtggtctcc ccagtttatt ctgcttctcc tagcagttac aacctacgga
     1861 tttccactcg aggagaaatt cgatgggctc ttccgtgcag agccaccaca ctgttccaag
     1921 acaccaattg tccgtgctca aacatctcaa aatgcgatgt catcaattgc aagaggaatg
     1981 caaatgcaat tctcaattgg tttgcacact gcagtctgct tcagattata tgaagacact
     2041 caactcgcat cccaagagat caatgatgat gaaaatgctg gaaatcagac atcacttctt
     2101 catactattc gattggaaaa gcttgaacat caccatccga taactcagcg gtacactttt
     2161 ggaatccctg aagttcatgc cagttgtatc tgtgagtgtg atgccacatc ttcaacatgt
     2221 accgcagaat ctcatcagtt taccgcttgt ccagagtctg ataagtctga tgaaacatct
     2281 tcatgctacc gcacgttctt cccaaatcag acaccaatcg ggtgcagtga agacgacatc
     2341 cccaagcttt gctgtgacgt tcgattcaag ccatataaaa atatgacatt cctcgcagtc
     2401 aaactagaac aacctacaac gtatgcaaca tttgtctacg cagcttatga ttttgtaaat
     2461 ggatattggg tggaaaaaga taaaactaaa atccgttctc aattagatgg tggaacacag
     2521 gataggcatc tcgaccaaaa acgtcgaatt tcattggctg taactgccgg aggacgagca
     2581 tctcatcaac tggagacagg aatgtatttc tcaagaacaa gtaatggagg agaaacagaa
     2641 gaactgagaa tgcaaccatt gaatgagatt acagacaata actttgaccg tcttggatgg
     2701 tacagaatgg acgattctgg tcattttcat gtgaataatg gagttgtgaa aatggatgat
     2761 attcacaagg ctaaagtgaa aaattgcaaa gagcaaactt ataaatcaat tctctctgca
     2821 aatcattata tgccaggaca tttcaatttg actcgtccac ttgaagttat caaaccgtgg
     2881 attcaatctg ctcgaatttt tgatagttct ctcagacaag ctgttgtaac acatgccgaa
     2941 ggaactaatc ttcaaatttc aattcacttg gatgacgagg tggaaagtca aaaccttgtc
     3001 ttcttccata atgcatcaag aattcgtgac ttcagtggat ccatcatcgt tgattcaaaa
     3061 tcaaatcgat tgttcaactt gacagtatat gaggcttctg gaaaaattga tggatcagtg
     3121 aagatgtcaa ctggatttgg atctgataca attcacacat tcactgctta tgttagtgat
     3181 cttcatgctt caaaccgttc tatgattatt ccactgccag ccatcgttgg acaaggagcc
     3241 agagcaatct gtctccgagc agattcaatg gctgatattg acaaaatttg tcatgtcatt
     3301 gagtatttcg agagcccttt ggaaatcgat cttgttgagg gaaaatggca cgaaatgatt
     3361 ggaacctgcc caacttgtaa tcaaatcaat ttcaatggaa tgatgaaatt tctgaatcca
     3421 gctcattgga tcaaaggaat cagttcgatt ggtgacggcg ttatgattgc tacggatatt
     3481 gttgtgtatc tcggagtttt gtgtattctg tatcttctta ttacgaagat tattgttccg
     3541 ttggttcgtt gctgggtatg ccccatgtcg attttctgca atgggtcctc ttcgtcgtcc
     3601 aaaaacaaaa atgataagag aaggaaggag agagaggaga ggcgcagaaa ggacaaattt
     3661 gtctcagaat ccgaagatgg tgcaagaagt agttccgagc cccacgacac tttggcgaga
     3721 taccatggaa atcactcgga gcggcactat agcagtagcc agtacattgg tctagagggc
     3781 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgtgaccc
     3841 gggggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     3901 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3961 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4021 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     4081 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     4141 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     4201 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     4261 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     4321 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     4381 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     4441 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     4501 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4561 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4621 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4681 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4741 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4801 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4861 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4921 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4981 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     5041 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     5101 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta catgagcggc
     5161 ggcctgcaga agaagctgga ggagctggag ctggactaat gtacaagtaa agcggccggc
     5221 cgccagcaca gtggtcgatc gaccaatgcc ctggctcaca aataccactg agatcttttt
     5281 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     5341 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     5401 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     5461 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     5521 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     5581 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     5641 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     5701 cctcttctct tatggagatc cctcgacctg caccgtcgac cagctggtcg acggtgcacc
     5761 gtcgaccagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5821 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5881 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5941 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6061 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6121 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     6181 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6241 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     6301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8281 tg
//
