LOCUS       sequence-333504-        5328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rab5C fused to mRuby3 fluorescent protein (ex558/em592) tag in N-ter
            under control of a weak L30 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5328)
  TITLE     Generation of mRuby3-fused Rab GTPases constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5328)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBS L30 mRuby3-Rab6b
FEATURES             Location/Qualifiers
     source          1..5328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..53)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     153..172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      239..694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(326..345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     536..557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     820..842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     834..851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     835..851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     858..877
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        858..876
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1289..1298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1295..2005
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     protein_bind    2039..2063
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2694..2718)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2762..2781)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2806..2854
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(3070..3088)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3109..3125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3109..3125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3122..3144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3133..3149
                     /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(3157..3163,3164..3181,3182..3187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3202..3223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3340..3357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3511..4099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3591..3610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4270..5061,5062..5130))
                     /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     4893..4912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5131..5235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5303..5321
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
       61 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
      121 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      181 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggac
      241 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      301 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      361 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      421 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      481 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      541 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      601 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      661 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
      721 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      781 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      841 cgacggccag tgaattgtaa tacgactcac tatagggcga attcgagctc ggtacccggg
      901 gatcgcagaa caaacgccca gaaccgaagg ccggagccca gacgccgcag cagcccagcc
      961 tgcgttgtaa cccggtcccc atagcaacca actaccgcag actactctcg caagcgctgt
     1021 tctgcttgtg acccggaccg gcttctcttc cgcttccggt cccacaatcc tctgtcggcc
     1081 tagaagagct ttgcattgtg ggagctcctt cctttctcgc tccccggcca tcttggcggc
     1141 tggtgttggt gagtgagctc tgcggggtaa acgattaggc ggctcgggga gctccgctag
     1201 ctggtgtttg acgctctgga ttataatcct ctcttctgtc ctctgtgtat gctaggttgg
     1261 gggttgtccc gcacctacta gaggatccgc caccatggtg tctaagggcg aagagctgat
     1321 caaggaaaat atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc accaattcaa
     1381 atgcacaggt gaaggagaag gcagaccgta cgagggagtg caaaccatga ggatcaaagt
     1441 catcgaggga ggacccctgc catttgcctt tgacattctt gccacgtcgt tcatgtatgg
     1501 cagccgtacc tttatcaagt acccggccga catccctgat ttctttaaac agtcctttcc
     1561 tgagggtttt acttgggaaa gagttacgag atacgaagat ggtggagtcg tcaccgtcac
     1621 gcaggacacc agccttgagg atggcgagct cgtctacaac gtcaaggtca gaggggtaaa
     1681 ctttccctcc aatggtcccg tgatgcagaa gaagaccaag ggttgggagc ctaatacaga
     1741 gatgatgtat ccagcagatg gtggtctgag aggatacact gacatcgcac tgaaagttga
     1801 tggtggtggc catctgcact gcaacttcgt gacaacttac aggtcaaaaa agaccgtcgg
     1861 gaacatcaag atgcccggtg tccatgccgt tgatcaccgc ctggaaagga tcgaggagag
     1921 tgacaatgaa acctacgtag tgcaaagaga agtggcagtt gccaaataca gcaaccttgg
     1981 tggtggcatg gacgagctgt acaagggtgg ttccggtgga tctggaggtt ctgatatcac
     2041 aagtttgtac aaaaaagcag gcaccatgtc cgcaggggga gattttggga atccactgag
     2101 aaaattcaag ttggtgttct tgggggagca gagcgtcggg aagacgtctc tgattacgag
     2161 gttcatgtac gacagcttcg acaacacata ccaggcaacc attgggattg acttcttgtc
     2221 aaaaaccatg tacttggagg accgcacggt gcgactgcag ctctgggaca cagctggtca
     2281 ggagaggttc cgcagcctga tccccagcta catccgggac tccacggtgg ctgtggtggt
     2341 gtacgacatc acaaatctca actccttcca acagacctct aagtggatcg acgacgtcag
     2401 gacagagagg ggcagtgatg ttatcatcat gctggtgggc aacaagacgg acctggctga
     2461 taagaggcag ataaccatcg aggaggggga gcagcgcgcc aaagaactga gcgtcatgtt
     2521 cattgagacc agtgcgaaga ctggctacaa cgtgaagcag ctttttcgac gtgtggcgtc
     2581 ggctctaccc ggaatggaga atgtccagga gaaaagcaaa gaagggatga ttgacatcaa
     2641 gctggacaaa ccccaggagc ccccggccag cgagggcggc tgctcctgct agaacccagc
     2701 tttcttgtac aaagtggtga tatcgtaggt tgacctgcag ggggagatgg gggaggctaa
     2761 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     2821 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     2881 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     2941 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     3001 gcggcaagcc ctgccatagc cacgggcccc gtgggttagg gacggggtcc cccatgcaag
     3061 cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt
     3121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3181 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5281 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcg
//
