LOCUS       sequence-268082-        5323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rab6A fused to the red fluorescent protein Ruby3 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 5323)
  AUTHORS   Deffieu MS, Cesonyte I, Delalande F, Boncompain G, Dorobantu C, Song
            E, Lucansky V, Hirschler A, Cianferani S, Perez F, Carapito C, 
            Gaudin R
  TITLE     Rab7-harboring vesicles are carriers of the transferrin receptor 
            through the biosynthetic secretory pathway.
  JOURNAL   Sci Adv. 2021 Jan 8;7(2). pii: 7/2/eaba7803. doi: 
            10.1126/sciadv.aba7803. Print 2021 Jan.
  PUBMED    33523982
REFERENCE   2  (bases 1 to 5323)
  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: "Sci Adv."; 
            date: "2021-01-8"; volume: "7(2). pii: 7/2/eaba7803. doi"; pages: " 
            10.1126/sciadv.aba7803. Print 2021 Jan"
COMMENT     Sequence Label: pBS L30 Ruby3-Rab6A
FEATURES             Location/Qualifiers
     source          1..5323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    741..765
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1391..1415)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1459..1478)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1503..1551
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(1767..1785)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1806..1822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1806..1822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1819..1841)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1830..1846
                     /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(1854..1860,1861..1878,1879..1884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1899..1920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2037..2054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2208..2796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2288..2307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2967..3758,3759..3827))
                     /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     3590..3609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3828..3932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4000..4018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4056..4078)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4178..4197
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4264..4719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4351..4370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4561..4582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4845..4867
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4859..4876
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4860..4876
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4883..4902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4883..4901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5314..5323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5320..5323,1..707)
                     /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"
ORIGIN
        1 tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt
       61 cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcagaccg tacgagggag
      121 tgcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc
      181 ttgccacgtc gttcatgtat ggcagccgta cctttatcaa gtacccggcc gacatccctg
      241 atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag
      301 atggtggagt cgtcaccgtc acgcaggaca ccagccttga ggatggcgag ctcgtctaca
      361 acgtcaaggt cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca
      421 agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg agaggataca
      481 ctgacatcgc actgaaagtt gatggtggtg gccatctgca ctgcaacttc gtgacaactt
      541 acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tgtccatgcc gttgatcacc
      601 gcctggaaag gatcgaggag agtgacaatg aaacctacgt agtgcaaaga gaagtggcag
      661 ttgccaaata cagcaacctt ggtggtggca tggacgagct gtacaagggt ggttccggtg
      721 gatctggagg ttctgatatc acaagtttgt acaaaaaagc aggcttctcc acgggcggag
      781 acttcgggaa tccgctgagg aaattcaagc tggtgttcct gggggagcaa agcgttggaa
      841 agacatcttt gatcaccaga ttcatgtatg acagttttga caacacctat caggcaacaa
      901 ttggcattga ctttttatca aaaactatgt acttggagga tcgaacaatc aggcttcagc
      961 tgtgggatac tgcgggtcag gaacgtttcc gtagcctcat tcccagttac atccgtgatt
     1021 ctgctgcagc tgtagtagtt tacgatatca caaatgttaa ctcattccag caaactacaa
     1081 agtggattga tgatgtcaga acagaaagag gaagtgatgt tatcatcatg ctagtaggaa
     1141 ataaaacaga tcttgctgac aagaggcaag tgtcaattga ggagggagag aggaaagcca
     1201 aagagctgaa tgttatgttt attgaaacta gtgcaaaagc tggatacaat gtaaagcagc
     1261 tctttcgacg tgtagcagca gctttgccgg gaatggaaag cacacaggac agaagcagag
     1321 aagatatgat tgacataaaa ctggaaaagc ctcaggagca accagtcagt gaaggaggct
     1381 gttcctgcta acccagcttt cttgtacaaa gtggtgatat cgtaggttga cctgcagggg
     1441 gagatggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     1501 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
     1561 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     1621 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     1681 tcgcagccaa cgtcggggcg gcaagccctg ccatagccac gggccccgtg ggttagggac
     1741 ggggtccccc atgcaagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
     1801 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1861 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3961 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4021 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4081 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4141 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4201 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4261 aggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4681 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     4741 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4801 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     4861 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattcg agctcggtac
     4921 ccggggatcg cagaacaaac gcccagaacc gaaggccgga gcccagacgc cgcagcagcc
     4981 cagcctgcgt tgtaacccgg tccccatagc aaccaactac cgcagactac tctcgcaagc
     5041 gctgttctgc ttgtgacccg gaccggcttc tcttccgctt ccggtcccac aatcctctgt
     5101 cggcctagaa gagctttgca ttgtgggagc tccttccttt ctcgctcccc ggccatcttg
     5161 gcggctggtg ttggtgagtg agctctgcgg ggtaaacgat taggcggctc ggggagctcc
     5221 gctagctggt gtttgacgct ctggattata atcctctctt ctgtcctctg tgtatgctag
     5281 gttgggggtt gtcccgcacc tactagagga tccgccacca tgg
//
