LOCUS       sequence-429029-        8123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split GFP assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8123)
  AUTHORS   Kodaka M, Yang Z, Nakagawa K, Maruyama J, Xu X, Sarkar A, Ichimura 
            A, Nasu Y, Ozawa T, Iwasa H, Ishigami-Yuasa M, Ito S, Kagechika H, 
            Hata Y
  TITLE     A new cell-based assay to evaluate myogenesis in mouse myoblast 
            C2C12 cells.
  JOURNAL   Exp Cell Res. 2015 Aug 15;336(2):171-81. doi: 
            10.1016/j.yexcr.2015.06.015. Epub 2015 Jun 24.
  PUBMED    26116467
REFERENCE   2  (bases 1 to 8123)
  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.yexcr.2015.06.015"; journalName: "Exp Cell Res"; date: 
            "2015-08-15- 15"; volume: "336"; issue: "2"; pages: "171-81"
COMMENT     Sequence Label: pQCXIP-GFP1-10
FEATURES             Location/Qualifiers
     source          1..8123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(400..858,859..861,862..864))
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GKYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITA"
     primer_bind     798..819
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(985..1009)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1006..1209)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1210..1513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(1550..1966)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1566..1582)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1598..1620)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2167..2366
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             2429..2604
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     promoter        complement(2605..2808)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        2809..3188
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3288..3306)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3374..3478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3479..3547,3548..4339)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3697..3716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4510..5098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4999..5018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5252..5269
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5350..5369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(5391..5720)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5405..5540
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5705..5725
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     5861..5883
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     LTR             5986..6411
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     CDS             complement(6785..7384)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6868..6888)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7365..7384
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    7417..7879
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7577..7596)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     7806..7823
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 ggttgtgtag tgaatggccc tttccatctg tgtgggcgac gtaccagccc gctcgccagt
       61 ggggcgcgcc ttcaatgtgg agccctctgg attagggcag tgaagcggca atcgggacgc
      121 gcaggcaacg gtgggggacg gggcgagctg ggcggggtag ggaggggcga tacacggcgg
      181 ggagaggcga ggaacaagag gaggcgggcg gaattcctac ttctcgttgg ggtctttgct
      241 caggacggtc tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
      301 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga cgttgtggcg
      361 gacggtgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg
      421 gctgttgaag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcggt
      481 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacca cggcgcgggt
      541 cttgtatttg ccgtcgtcct tgaaggagat ggtgcgctcc tggacgtagc cttcgggcat
      601 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
      661 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
      721 gatgaacttc aaggtcagct tgccgatggt ggcatcgccc tcgccctcgc cgcgcacgct
      781 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
      841 cagctcctcg cccttgctca ccatggatcg gccgcggatc aattcctgca ggtccgttcc
      901 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
      961 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     1021 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     1081 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     1141 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     1201 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     1261 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     1321 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     1381 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     1441 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     1501 aagttatgta acggactcta gaggatcgat cccccagatc ttcaattccg gcgcctagag
     1561 aaggagtgag ggctggataa agggaggatc gaggcggggt cgaacgagga ggttcaaggg
     1621 ggagagacgg ggcggatgga ggaagaggag gcggaggctt agggtgtaca aagggcttga
     1681 cccagggagg ggggtcaaaa gccaaggctt cccaggtcac gatgtagggg acctggtctg
     1741 ggtgtccatg cgggccaggt gaaaagacct tgatcttaac ctgggtgatg aggtctcggt
     1801 taaaggtgcc gtctcgcggc catccgacgt taaaggttgg ccattctgca gagcagaagg
     1861 taacccaacg tctcttcttg acatctaccg actggttgtg agcgatccgc tcgacatctt
     1921 tccagtgacc taaggtcaaa cttaagggag tggtaacagt ctggccctaa ttttcagaca
     1981 aatacagaaa cacagtcaga cagagacaac acagaacgat gctgcagcgc tgcagcagac
     2041 aagacgcgcg gcttcggttc caaaccgaaa gcaaaaattc agacggaggc gggaactgtt
     2101 ttaggttctc gtctcctacc agaaccacat atcctgacgg ggtcggattc cacatcgact
     2161 cccttcctca ggtcgggcca caaaaacggc ccccaaagtc cctgggacgt ctcccagggt
     2221 tgcggccggg tgttcagaac tcgtcagttc caccacgggt ccgccagata cagagctagt
     2281 tagctaacta gtacagacgc aggcgcataa catcaaacat agacactaga caatcggaca
     2341 gacacagata agttgctggc cagcttacct cccggtggtg ggtcggtggt ccctgggcag
     2401 gggtctccaa atcccggacg agcccccaaa tgaaagaccc ccgtcgtggg tagtcaatca
     2461 ctcagaggag accctcccaa ggaacagcga gaccacgatt cggatgcaaa cagcaagagg
     2521 ctttattggg aatacgggta cccgggcgac gcagtctatc ggaagactgg cgcgccgagt
     2581 gaggggttgt gggctctttt attgagctct gcttatatag acctcccacc gtacacgcct
     2641 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     2701 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     2761 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     2821 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     2881 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     2941 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     3001 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     3061 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     3121 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     3181 tcaatgtcaa catggcggta atgttggaca tgagccaata taaatgtaca tattatgata
     3241 tggatacaac gtatgcaatg gccaagctaa ttcttgaaga cgaaagggcc tcgtgatacg
     3301 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3361 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3661 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3721 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3781 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4021 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4081 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4321 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4621 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4861 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4921 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4981 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5041 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5101 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5161 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5221 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5281 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtaagctt tttgcaaaag
     5401 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     5461 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     5521 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     5581 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     5641 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     5701 ccacacccta actgacacac attccacagc cggatctaca ctccgctatc gctacgtgac
     5761 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5821 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5881 aggttttcac cgtcatcacc gaaacgcgcg aggcagccag cttacctccc ggtggtgggt
     5941 cggtggtccc tgggcagggg tctcccgatc ccggacgagc ccccaaatga aagacccccg
     6001 ctgacgggta gtcaatcact cagaggagac cctcccaagg aacagcgaga ccacaagtcg
     6061 gatgcaactg caagagggtt tattggatac acgggtaccc gggcgactca gtcaatcgga
     6121 ggactggcgc cccgagtgag gggttgtggg ctcttttatt gagctcgggg agcagaagcg
     6181 cgcgaacaga agcgagaagc gaactgattg gttagttcaa ataaggcaca gggtcatttc
     6241 aggtccttgg ggcaccctgg aaacatctga tggttctcta gggtcgaccc tgttccatct
     6301 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
     6361 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
     6421 ttctggagac taaataaaat cttttatttt atctatggct cgtactctat aggcttcagc
     6481 tggtgatatt gttgagtcaa aactagagcc tggaccactg atatcctgtc tttaacaaat
     6541 tggactaatc gataagctta tctctacact ggctcgatgg agtcagccta gtgtccctcg
     6601 gaaagccttg aggtcccttt ggagcagttc tgcactcggc ctctgcagcg ctgcagccct
     6661 cgactagagt cggtgggcct cgggggcggg tgcggggtcg gcggggccgc cccgggtggc
     6721 ttcggtcgga gccatggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg
     6781 ggcgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
     6841 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
     6901 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
     6961 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
     7021 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
     7081 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
     7141 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
     7201 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
     7261 ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
     7321 tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
     7381 tcatggaagg tcgtctcctt gtgggttgtg gcaagcttat catcgtgttt ttcaaaggaa
     7441 aaccacgtcc ccgtggttcg gggggcctag acgttttttt aacctcgact aaacacatgt
     7501 aaagcatgtg caccgaggcc ccagatcaga tcccatacaa tggggtacct tctgggcatc
     7561 cttcagcccc ttgttgaata cgcttgagga gagccatttg actctttcca caactatcca
     7621 actcacaacg tggcactggg gttgtgccgc ctttgcaggt gtatcttata cacgtggctt
     7681 ttggccgcag aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa gggtcgctac
     7741 agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga cattcaacag
     7801 accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg
     7861 gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata
     7921 gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag ggggggctct
     7981 ctcttataca cctctgccgc tgcccactac tattgctgtc tctttctcgc ttcttccttt
     8041 ctctctccga aacatcttcc tcttttcttc tctctcttct cttccttctt ctctacttct
     8101 ccttctctac cttccttttc tcc
//
