LOCUS       sequence-325125-        5491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of zebrafish GFP-tagged Rab13 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5491)
  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 5491)
  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-Rab13 in pCSDEST2
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        984..1002
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     984..1001
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1057..1081
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1082..1091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1088..1090,1091..1093,1094..1804)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1088..1090,1091..1093,1094..1804)
                     /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(1133..1154)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1394..1413)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1741..1762
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1806..1830)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2432..2452)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2492..2626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2516..2535)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2570..2589
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2739..2759)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2739..2757)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2778..2794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2778..2794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2791..2813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2802..2818
                     /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(2826..2832,2833..2850,2851..2856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2871..2892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3009..3026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3180..3768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3260..3279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3939..4730,4731..4799))
                     /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     4562..4581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4800..4904)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4930..5385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5067..5088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5279..5298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5463..5491,1..949)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg
       61 tgccaattca atatggtgga tctggacccc agccaattca atatggcgga cttggcacca
      121 tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg
      181 gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg
      241 gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg caatatccaa
      301 tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc
      361 atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt
      421 cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca
      481 atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat
      541 ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga
      601 cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg
      661 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
      721 tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca
      781 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
      841 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
      901 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc
      961 ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct
     1021 ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa aaaagcaggc
     1081 tgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1141 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1201 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1261 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1321 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1381 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1441 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1501 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1561 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1621 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1681 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1741 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1801 caagcaccca gctttcttgt acaaagtggt ggcaaagaag tacgacttcc tctttaaatt
     1861 attgcttatc ggcgatagcg gtgttggcaa gacatgtttg atcatccgtt tcgcagagga
     1921 caattttaat tcaacgtaca tctcaaccat cggtattgac ttcaaagtga agaccattga
     1981 ggtggaagga aagaaagtga agctgcaagt ctgggacaca gcaggacagg agaggtttaa
     2041 gaccatcacc actgcatact acagaggggc catgggaatc attctcgtgt atgacataac
     2101 tgatgaaaaa tcctatgaga atattcagaa ctggatgaag agcatcaaag aaaatgcatc
     2161 agcgggtgta agtcgaatgt tactgggtaa caagtgtgac attgaggcca agaggaaagt
     2221 ctcaaaagag actggtgaga agctcgcaaa ggaacatggt attcgattct ttgagactag
     2281 tgccaaatcg agcatcaacg ttgaggagtc ttttacttct cttgctcgag acattttgct
     2341 gaagtcaaat aagaagccgg gtccatctgg acgtgaggtc aaacttacca gcacagagaa
     2401 aaagtcctcc tccaaatgtc ttcttcttta gcaactttat tatacatagt tgcactcgag
     2461 cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga
     2521 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2581 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2641 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc
     2701 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     2761 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2821 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2881 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2941 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3001 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3061 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3121 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3181 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3241 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3301 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3361 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3421 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3481 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3541 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3601 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3661 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3721 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3781 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4021 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4081 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4141 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4201 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4261 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4321 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4381 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4441 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4501 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4561 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4621 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4681 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4741 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4801 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4861 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4921 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4981 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5041 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5101 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5161 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5221 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5281 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5341 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     5401 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga
     5461 ccgccaattc aatatggcgt atatggactc a
//
