LOCUS       sequence-448190-        7651 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 7651)
  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 7651)
  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-BSR-GFP11
FEATURES             Location/Qualifiers
     source          1..7651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     428..447
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    join(467..475,476..478,479..1053)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(640..659)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     869..886
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(1084..1107)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1108..1155)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             complement(1168..1563)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(1699..1723)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1720..1923)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1924..2227
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2264..2680)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2280..2296)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2312..2334)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2881..3080
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3143..3318
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     promoter        complement(3319..3522)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3523..3902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4002..4020)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4088..4192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4193..4261,4262..5053)
                     /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(4411..4430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5224..5812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5713..5732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5966..5983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6064..6083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(6105..6434)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6119..6254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6419..6439
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     6575..6597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     LTR             6700..7125
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     CDS             complement(join(7499..7651,1..447))
                     /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(7582..7602)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac
       61 gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc
      121 catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc
      181 ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc
      241 gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg
      301 cagctcggtg acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta
      361 gtcggcgaac gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag
      421 gcgcaccgtg ggcttgtact cggtcatgga aggtcgtctc cttgtgggtt gtggcaagct
      481 tatcatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc tagacgtttt
      541 tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc agatcccata
      601 caatggggta ccttctgggc atccttcagc cccttgttga atacgcttga ggagagccat
      661 ttgactcttt ccacaactat ccaactcaca acgtggcact ggggttgtgc cgcctttgca
      721 ggtgtatctt atacacgtgg cttttggccg cagaggcacc tgtcgccagg tggggggttc
      781 cgctgcctgc aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc cagaggaact
      841 gcttccttca cgacattcaa cagaccttgc attcctttgg cgagagggga aagaccccta
      901 ggaatgctcg tcaagaagac agggccaggt ttccgggccc tcacattgcc aaaagacggc
      961 aatatggtgg aaaataacat atagacaaac gcacaccggc cttattccaa gcggcttcgg
     1021 ccagtaacgt tagggggggg ggagggagag gggggggggg gggggggggg gaattcgata
     1081 tcacttgtcg tcgtcgtcct tgtagtcagt gatcccggcg gcgttcacgt actcgtgcag
     1141 gaccatgtga tcgcgctgca gggatccgcc ctcccacaca taaccagagg gcagcaattc
     1201 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     1261 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     1321 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     1381 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     1441 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     1501 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     1561 catggtttag ttcctcacct tgcggccgcg gatcaattcc tgcaggtccg ttccaatgca
     1621 ccgttcccgg ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg
     1681 tggatggcgt ctccaggcga tctgacggtt cactaaacga gctctgctta tatagacctc
     1741 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     1801 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     1861 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     1921 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     1981 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     2041 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     2101 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     2161 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     2221 tgtaacggac tctagaggat cgatccccca gatcttcaat tccggcgcct agagaaggag
     2281 tgagggctgg ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag
     2341 acggggcgga tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg
     2401 gaggggggtc aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc
     2461 catgcgggcc aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg
     2521 tgccgtctcg cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc
     2581 aacgtctctt cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt
     2641 gacctaaggt caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca
     2701 gaaacacagt cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg
     2761 cgcggcttcg gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt
     2821 tctcgtctcc taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc
     2881 ctcaggtcgg gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc
     2941 cgggtgttca gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta
     3001 actagtacag acgcaggcgc ataacatcaa acatagacac tagacaatcg gacagacaca
     3061 gataagttgc tggccagctt acctcccggt ggtgggtcgg tggtccctgg gcaggggtct
     3121 ccaaatcccg gacgagcccc caaatgaaag acccccgtcg tgggtagtca atcactcaga
     3181 ggagaccctc ccaaggaaca gcgagaccac gattcggatg caaacagcaa gaggctttat
     3241 tgggaatacg ggtacccggg cgacgcagtc tatcggaaga ctggcgcgcc gagtgagggg
     3301 ttgtgggctc ttttattgag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     3361 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     3421 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     3481 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     3541 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     3601 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     3661 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     3721 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     3781 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     3841 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     3901 tcaacatggc ggtaatgttg gacatgagcc aatataaatg tacatattat gatatggata
     3961 caacgtatgc aatggccaag ctaattcttg aagacgaaag ggcctcgtga tacgcctatt
     4021 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4081 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4141 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4201 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4261 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4321 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4381 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4441 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4501 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4561 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4621 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4681 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     4741 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4801 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4861 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4921 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4981 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5041 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5101 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5161 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5221 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5281 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5341 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5401 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5461 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5521 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5581 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5641 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5701 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5761 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5821 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5881 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5941 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     6001 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     6061 cagtacaatc tgctctgatg ccgcatagtt aagccagtaa gctttttgca aaagcctagg
     6121 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     6181 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     6241 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     6301 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     6361 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     6421 cctaactgac acacattcca cagccggatc tacactccgc tatcgctacg tgactgggtc
     6481 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6541 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6601 tcaccgtcat caccgaaacg cgcgaggcag ccagcttacc tcccggtggt gggtcggtgg
     6661 tccctgggca ggggtctccc gatcccggac gagcccccaa atgaaagacc cccgctgacg
     6721 ggtagtcaat cactcagagg agaccctccc aaggaacagc gagaccacaa gtcggatgca
     6781 actgcaagag ggtttattgg atacacgggt acccgggcga ctcagtcaat cggaggactg
     6841 gcgccccgag tgaggggttg tgggctcttt tattgagctc ggggagcaga agcgcgcgaa
     6901 cagaagcgag aagcgaactg attggttagt tcaaataagg cacagggtca tttcaggtcc
     6961 ttggggcacc ctggaaacat ctgatggttc tctagggtcg accctgttcc atctgttcct
     7021 gaccttgatc tgaacttctc tattctcagt tatgtatttt tccatgcctt gcaaaatggc
     7081 gttacttaag ctagcttgcc aaacctacag gtggggtctt tcattccccc ctttttctgg
     7141 agactaaata aaatctttta ttttatctat ggctcgtact ctataggctt cagctggtga
     7201 tattgttgag tcaaaactag agcctggacc actgatatcc tgtctttaac aaattggact
     7261 aatcgataag cttatctcta cactggctcg atggagtcag cctagtgtcc ctcggaaagc
     7321 cttgaggtcc ctttggagca gttctgcact cggcctctgc agcgctgcag ccctcgacta
     7381 gagtcggtgg gcctcggggg cgggtgcggg gtcggcgggg ccgccccggg tggcttcggt
     7441 cggagccatg gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc
     7501 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     7561 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
     7621 ggaaggcggg caccccggcg cgctcggccg c
//
