LOCUS       sequence-333545-        5322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rab18 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 5322)
  TITLE     Generation of mRuby3-fused Rab GTPases constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5322)
  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            "Unpublished"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-Rab18
FEATURES             Location/Qualifiers
     source          1..5322
                     /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(2688..2712)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2756..2775)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2800..2848
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(3064..3082)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3103..3119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3103..3119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3116..3138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3127..3143
                     /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(3151..3157,3158..3175,3176..3181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3196..3217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3334..3351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3505..4093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3585..3604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4264..5055,5056..5124))
                     /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     4887..4906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5125..5229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5297..5315
                     /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 gcaccatgga cgaggacgtg ctaaccaccc tgaagatcct
     2101 catcatcggc gagagtgggg tgggcaagtc cagcctgctc ttgaggttca cagatgatac
     2161 gtttgatcca gaacttgcag caacaatagg tgttgacttt aaggtgaaaa caatttcagt
     2221 ggatggaaat aaggctaaac ttgcaatatg ggatactgct ggtcaagaga ggtttagaac
     2281 attaactccc agctattata gaggtgcaca gggtgttata ttagtttatg atgtcacaag
     2341 aagagataca tttgttaaac tggataattg gttaaatgaa ttggaaacat actgtacaag
     2401 aaatgacata gtaaacatgc tagttggaaa taaaatcgat aaggaaaatc gtgaagtcga
     2461 tagaaatgaa ggcctgaaat ttgcacgaaa gcattccatg ttatttatag aggcaagtgc
     2521 aaaaacctgt gatggtgtac aatgtgcctt tgaagaactt gttgaaaaga tcattcagac
     2581 ccctggactg tgggaaagtg agaaccagaa taaaggagtc aaactgtcac acagggaaga
     2641 aggccaagga ggaggagcct gtggtggtta ttgctctgtg ttatagaacc cagctttctt
     2701 gtacaaagtg gtgatatcgt aggttgacct gcagggggag atgggggagg ctaactgaaa
     2761 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2821 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     2881 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2941 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     3001 agccctgcca tagccacggg ccccgtgggt tagggacggg gtcccccatg caagcttttg
     3061 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     3121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cg
//
