LOCUS       sequence-448630-        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-GAPDH-CC fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  AUTHORS   Chen X, Fansler MM, Janjos U, Ule J, Mayr C
  TITLE     The FXR1 network acts as signaling scaffold for actomyosin 
            remodeling.
  JOURNAL   Cell. 5 August 2024. doi: 10.1016/j.cell.2024.07.015.
  PUBMED    37961296
REFERENCE   2  (bases 1 to 7166)
  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"; journalName: "Cell. 5 
            August 2024. doi: 10.1016/j.cell.2024.07.015."
COMMENT     Sequence Label: pcDNA3puro-EGFP-GAPDH-CC
FEATURES             Location/Qualifiers
     source          1..7166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(905..907,908..910,911..1621)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(950..971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1211..1230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3006..3023)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3012..3236
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3282..3710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3369..3388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3579..3600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3719..3739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3724..4053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3904..4039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3966..3985
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4066..4665
                     /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(4066..4085)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4562..4582
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4839..4960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4876..4895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4930..4949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5009..5025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5009..5025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5022..5044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5033..5049
                     /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(5057..5063,5064..5081,5082..5087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5102..5123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5240..5257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5411..5999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5491..5510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6170..6961,6962..7030))
                     /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     6793..6812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7031..7135)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      961 ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1381 gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1501 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 ggggaaggtg aaggtcggag tcaacggatt tggtcgtatt gggcgcctgg tcaccagggc
     1681 tgcttttaac tctggtaaag tggatattgt tgccatcaat gaccccttca ttgacctcaa
     1741 ctacatggtt tacatgttcc aatatgattc cacccatggc aaattccatg gcaccgtcaa
     1801 ggctgagaac gggaagcttg tcatcaatgg aaatcccatc accatcttcc aggagcgaga
     1861 tccctccaaa atcaagtggg gcgatgctgg cgctgagtac gtcgtggagt ccactggcgt
     1921 cttcaccacc atggagaagg ctggggctca tttgcagggg ggagccaaaa gggtcatcat
     1981 ctctgccccc tctgctgacg cccccatgtt cgtcatgggt gtgaaccatg agaagtatga
     2041 caacagcctc aagatcatca gcaatgcctc ctgcaccacc aactgcttag cacccctggc
     2101 caaggtcatc catgacaact ttggtatcgt ggaaggactc atgaccacag tccatgccat
     2161 cactgccacc cagaagactg tggatggccc ctccgggaaa ctgtggcgtg atggccgcgg
     2221 ggctctccag aacatcatcc ctgcctctac tggcgctgcc aaggctgtgg gcaaggtcat
     2281 ccctgagctg aacgggaagc tcactggcat ggccttccgt gtccccactg ccaacgtgtc
     2341 agtggtggac ctgacctgcc gtctagaaaa acctgccaaa tatgatgaca tcaagaaggt
     2401 ggtgaagcag gcgtcggagg gccccctcaa gggcatcctg ggctacactg agcaccaggt
     2461 ggtctcctct gacttcaaca gcgacaccca ctcctccacc tttgacgctg gggctggcat
     2521 tgccctcaac gaccactttg tcaagctcat ttcctggtat gacaacgaat ttggctacag
     2581 caacagggtg gtggacctca tggcccacat ggcctccaag gaggaacctg tcaagcgtgg
     2641 taatgacaca gatgtgcgga gaaaagagaa ggagaataga aagctacata tggagcttaa
     2701 atctgaacgt gagaaattga cccagcagat gatcaagtat cagaaagaac tgaatgaaat
     2761 gcaggcacaa atagctgaag agagccagat tcgaattgaa ctgcagatga cattggacag
     2821 taaagacagt gacattgagc agctgcggtc acaactccaa gccttgcata ttggtctgga
     2881 tagttccagt ataggcagtg gaccagggga tgctgaggca gatgatgggt ttccataatt
     2941 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
     3001 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3061 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3121 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3181 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3241 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3301 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3361 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3421 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3481 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3541 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3601 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3661 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3721 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3841 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3901 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3961 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4021 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc caaccatgac cgagtacaag
     4081 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4141 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4201 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4261 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4321 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4381 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4441 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4501 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4561 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4621 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagcggg actctggggt
     4681 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4741 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4801 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4861 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4921 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4981 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5041 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5101 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5161 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5221 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5281 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5341 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5401 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5461 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5521 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5581 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5641 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5701 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5761 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5821 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5881 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5941 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6001 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6061 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6121 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6181 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6241 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6301 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6361 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6421 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6481 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6541 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6601 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6661 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6721 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6781 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6841 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6901 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6961 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7021 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7081 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7141 ttccccgaaa agtgccacct gacgtc
//
