LOCUS       sequence-326580-        5731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of zebrafish GFP-tagged Rab19 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5731)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 5731)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: EGFP-Rab19
FEATURES             Location/Qualifiers
     source          1..5731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..1038
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1073..1091
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1073..1090
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1146..1170
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1171..1180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1177..1179,1180..1182,1183..1893)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1177..1179,1180..1182,1183..1893)
                     /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(1222..1243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1483..1502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1830..1851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1895..1919)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2761..2781)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2821..2955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2845..2864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2899..2918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3068..3088)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3068..3086)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3107..3123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3107..3123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3120..3142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3131..3147
                     /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(3155..3161,3162..3179,3180..3185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3200..3221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3338..3355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3509..4097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3589..3608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4268..5059,5060..5128))
                     /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     4891..4910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5129..5233)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5259..5714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5396..5417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5608..5627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
       61 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
      121 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      181 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      241 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      301 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      361 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      421 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      481 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      541 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      601 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      661 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      721 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      781 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      841 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      901 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      961 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
     1021 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1081 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcaag
     1141 gatcaacaag tttgtacaaa aaagcaggct gccaccatgg tgagcaaggg cgaggagctg
     1201 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1261 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1321 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1381 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1441 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1501 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1561 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1621 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1681 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1741 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1801 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1861 gggatcactc tcggcatgga cgagctgtac aagcacccag ctttcttgta caaagtggtg
     1921 ctgttctacc gagcaaaggt cacctttcaa gacagcactg tttgtgctcg atttcactcg
     1981 tactttaaaa actgtttatc tacagcagac agggaaagac tggggttttc tcctagcatt
     2041 agctttgtgt gtgtttgtgt ttgtgcagac ggagatgcag tggtgcagga cgtctctttt
     2101 ccaggtatac agtccgcagg gcgagaggac gaggagcgct gtgattttct gttcaagatc
     2161 atcctgattg gggacagtaa tgtggggaag acctgtgtta tccagagctt caggtcagca
     2221 gaggtcactg aactgcagca caacactatt ggagtggatt ttaccgtgcg cagcatggat
     2281 gttgacggca ggagagtcaa gatgcaggtg tgggacacgg cgggacagga gcgtttccgc
     2341 accatcactc agagctatta tcgcagtgct catggcgcta tgattgcgta tgatctgagc
     2401 cgccgcgaca ccttcgactc tctgccacac tggattcagg cgctggagca gtacggagct
     2461 gccagcctcg tcctcgtact catcggtaat aagtgtgatc tggaggcgca gcggcaggtg
     2521 ctgttcgagg acgcctgtac gctagcggag cgcagtggtg ctctggccgc tttagaaaca
     2581 tccgccaaac accatcacaa catcgaggag gctttccaac tgatggccag agaactctta
     2641 gtgcgacacg gaggaataca ttatcaggac aaccaatcag attcacccac cgtctacctg
     2701 cactctgact cacacccaat cgaaggagaa cagttggaga aaaggtcatg tgactgctaa
     2761 caactttatt atacatagtt gcactcgagc ctctagaact atagtgagtc gtattacgta
     2821 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2881 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2941 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3001 gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct
     3061 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     3121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt
     3181 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3241 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3301 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3361 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3421 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3481 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3541 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3601 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3661 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3721 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3781 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3841 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3901 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3961 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4021 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4081 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4501 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4561 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4621 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4681 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4741 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4801 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4861 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4921 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4981 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5041 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5101 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5161 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5221 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5281 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5341 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5401 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5461 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5521 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5581 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5641 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5701 gctacagggc gcgtcccatt cgccattcag g
//
