LOCUS       sequence-346607-        5147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of EGFP. Parton lab clone DFS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5147)
  AUTHORS   Lo HP, Lim YW, Xiong Z, Martel N, Ferguson C, Ariotti N, Giacomotto 
            J, Rae J, Floetenmeyer M, Moradi SV, Gao Y, Tillu VA, Xia D, Wang H,
            Rahnama S, Nixon SJ, Bastiani M, Day RD, Smith KA, Palpant NJ, 
            Johnston WA, Alexandrov K, Collins BM, Hall TE, Parton RG
  TITLE     Cavin4 interacts with Bin1 to promote T-tubule formation and 
            stability in developing skeletal muscle.
  JOURNAL   J Cell Biol. 2021 Dec 6;220(12). pii: 212693. doi: 
            10.1083/jcb.201905065. Epub 2021 Oct 11.
  PUBMED    34633413
REFERENCE   2  (bases 1 to 5147)
  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: "J Cell 
            Biol."; date: "2021-12-6"; volume: "220(12). pii: 212693. doi"; 
            pages: " 10.1083/jcb.201905065. Epub 2021 Oct 11"
COMMENT     Sequence Label: EGFP
FEATURES             Location/Qualifiers
     source          1..5147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(138..140,141..143,144..854)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(138..140,141..143,144..854)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(183..204)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(444..463)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     791..812
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(916..940)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    942..1076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1139..1159)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    1199..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1446..1466)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1446..1464)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1485..1501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1485..1501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1498..1520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1509..1525
                     /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(1533..1539,1540..1557,1558..1563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1578..1599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1716..1733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1887..2475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1967..1986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2646..3437,3438..3506))
                     /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     3269..3288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3507..3611)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3637..4092)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3774..3795)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3986..4005
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4163..5147
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctggaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      181 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      241 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      301 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      361 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      421 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      481 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      541 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
      601 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
      661 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
      721 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
      781 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
      841 acgagctgta caagggcggt ggaagatctg ggaattcaag gcctctcgag cctctagatt
      901 ctgcagccct atagcaccca gctttcttgt acaaagtggg ggatccagac atgataagat
      961 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1021 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1081 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcca
     1141 actttattat acatagttgc actcgagcct ctagaactat agtgagtcgt attacgtaga
     1201 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1261 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1321 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1381 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     1441 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     1501 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     1561 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1621 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1681 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1741 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1801 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1861 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1921 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1981 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2041 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2101 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2161 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2221 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2281 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2341 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2401 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2461 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2521 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2581 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2641 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2701 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2761 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2821 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2881 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2941 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3001 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3061 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3121 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3181 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3241 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3301 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3361 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3421 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3481 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3541 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3601 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     3661 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3721 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3781 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3841 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3901 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3961 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4021 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4081 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4141 gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg
     4201 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
     4261 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
     4321 ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt
     4381 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
     4441 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
     4501 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
     4561 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
     4621 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
     4681 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
     4741 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
     4801 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
     4861 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
     4921 tattaatagt aacttggcaa gtacattact attggaagta cgccagggta cattggcagt
     4981 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
     5041 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
     5101 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
