LOCUS       sequence-268092-        5325 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Rab7A 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 5325)
  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 5325)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Rab7A
FEATURES             Location/Qualifiers
     source          1..5325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      455..464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             461..1171
                     /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    1205..1229
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1857..1881)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1925..1944)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1969..2017
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(2233..2251)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2272..2288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2272..2288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2285..2307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2296..2312
                     /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(2320..2326,2327..2344,2345..2350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2365..2386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2503..2520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2674..3262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2754..2773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3433..4224,4225..4293))
                     /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     4056..4075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4294..4398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4466..4484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4522..4544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4644..4663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4730..5185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4817..4836)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5027..5048
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(5311..5325,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5325,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattc gagctcggta
       61 cccggggatc gcagaacaaa cgcccagaac cgaaggccgg agcccagacg ccgcagcagc
      121 ccagcctgcg ttgtaacccg gtccccatag caaccaacta ccgcagacta ctctcgcaag
      181 cgctgttctg cttgtgaccc ggaccggctt ctcttccgct tccggtccca caatcctctg
      241 tcggcctaga agagctttgc attgtgggag ctccttcctt tctcgctccc cggccatctt
      301 ggcggctggt gttggtgagt gagctctgcg gggtaaacga ttaggcggct cggggagctc
      361 cgctagctgg tgtttgacgc tctggattat aatcctctct tctgtcctct gtgtatgcta
      421 ggttgggggt tgtcccgcac ctactagagg atccgccacc atggtgtcta agggcgaaga
      481 gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca
      541 attcaaatgc acaggtgaag gagaaggcag accgtacgag ggagtgcaaa ccatgaggat
      601 caaagtcatc gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat
      661 gtatggcagc cgtaccttta tcaagtaccc ggccgacatc cctgatttct ttaaacagtc
      721 ctttcctgag ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac
      781 cgtcacgcag gacaccagcc ttgaggatgg cgagctcgtc tacaacgtca aggtcagagg
      841 ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa
      901 tacagagatg atgtatccag cagatggtgg tctgagagga tacactgaca tcgcactgaa
      961 agttgatggt ggtggccatc tgcactgcaa cttcgtgaca acttacaggt caaaaaagac
     1021 cgtcgggaac atcaagatgc ccggtgtcca tgccgttgat caccgcctgg aaaggatcga
     1081 ggagagtgac aatgaaacct acgtagtgca aagagaagtg gcagttgcca aatacagcaa
     1141 ccttggtggt ggcatggacg agctgtacaa gggtggttcc ggtggatctg gaggttctga
     1201 tatcacaagt ttgtacaaaa aagcaggcac catgacctct aggaagaaag tgttgctgaa
     1261 ggttatcatc ctgggagatt ctggagtcgg gaagacatca ctcatgaacc agtatgtgaa
     1321 taagaaattc agcaatcagt acaaagccac aataggagct gactttctga ccaaggaggt
     1381 gatggtggat gacaggctag tcacaatgca gatatgggac acagcaggac aggaacggtt
     1441 ccagtctctc ggtgtggcct tctacagagg tgcagactgc tgcgttctgg tatttgatgt
     1501 gactgccccc aacacattca aaaccctaga tagctggaga gatgagtttc tcatccaggc
     1561 cagtccccga gatcctgaaa acttcccatt tgttgtgttg ggaaacaaga ttgacctcga
     1621 aaacagacaa gtggccacaa agcgggcaca ggcctggtgc tacagcaaaa acaacattcc
     1681 ctactttgag accagtgcca aggaggccat caacgtggag caggcgttcc agacgattgc
     1741 acggaatgca cttaagcagg aaacggaggt ggagctgtac aacgaatttc ctgaacctat
     1801 caaactggac aagaatgacc gggccaaggc ctcggcagaa agctgcagtt gctagaaccc
     1861 agctttcttg tacaaagtgg tgatatcgta ggttgacctg cagggggaga tgggggaggc
     1921 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1981 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2041 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2101 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2161 ggggcggcaa gccctgccat agccacgggc cccgtgggtt agggacgggg tcccccatgc
     2221 aagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct
     2281 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2341 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2401 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2461 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2521 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2581 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4441 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4501 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4561 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4621 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4681 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga cgcgccctgt
     4741 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4801 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4861 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4921 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4981 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5041 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5101 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5161 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     5221 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     5281 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt
//
