LOCUS       sequence-469208-        3285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of His6-RAB1A Q70L 1-203 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3285)
  AUTHORS   Tremel S, Ohashi Y, Morado DR, Bertram J, Perisic O, Brandt LTL, von
            Wrisberg MK, Chen ZA, Maslen SL, Kovtun O, Skehel M, Rappsilber J, 
            Lang K, Munro S, Briggs JAG, Williams RL
  TITLE     Structural basis for VPS34 kinase activation by Rab1 and Rab5 on 
            membranes.
  JOURNAL   Nat Commun. 2021 Mar 10;12(1):1564. doi: 10.1038/s41467-021-21695-2.
  PUBMED    33692360
REFERENCE   2  (bases 1 to 3285)
  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: "2021-03-10"; volume: "12(1):1564. doi"; pages: " 
            10.1038/s41467-021-21695-2"
COMMENT     Sequence Label: pYO1262
FEATURES             Location/Qualifiers
     source          1..3285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..150
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     203..222
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        203..221
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             295..317
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             361..381
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1086..1104)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1100..1147
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1192..1210)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1278..1382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1383..1451,1452..2243)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1601..1620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2414..3002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2903..2922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3156..3173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(3188..3285,1..42)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3253..3272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
       61 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      121 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
      181 caggatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
      241 tctagtgccg gctccggaga gctccaattg gaattcgcta gctctagaaa taattttgtt
      301 taactttaag aaggagatat aaatatggct caccatcatc atcatcattc ttctggttcc
      361 gaaaacctgt atttccaggg ctcccatatg tccagcatga atcccgaata tgattattta
      421 ttcaagttac ttctgattgg cgactcaggg gttggaaagt cttgccttct tcttaggttt
      481 gcagatgata catatacaga aagctacatc agcacaattg gtgtggattt caaaataaga
      541 actatagagt tagacgggaa aacaatcaag cttcaaatat gggacacagc aggcctggaa
      601 agatttcgaa caatcacctc cagttattac agaggagccc atggcatcat agttgtgtat
      661 gatgtgacag atcaggagtc cttcaataat gttaaacagt ggctgcagga aatagatcgt
      721 tatgccagtg aaaatgtcaa caaattgttg gtagggaaca aatgtgatct gaccacaaag
      781 aaagtagtag actacacaac agcgaaggaa tttgctgatt cccttggaat tccgtttttg
      841 gaaaccagtg ctaagaatgc aacgaatgta gaacagtctt tcatgacgat ggcagctgag
      901 attaaaaagc gaatgggtcc cggagcaaca gctggtggtg ctgagaagtc caatgttaaa
      961 attcagagca ctccagtcaa gcagtcaggt ggaggttaag gatccagatc taagcttggt
     1021 accacgcgtg cgcgctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1081 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1141 ttttttgctg aaaggaggaa ctatagccgg ataattcttg aagacgaaag ggcctcgtga
     1201 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1261 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1321 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1381 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1441 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1501 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1561 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1621 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1681 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1741 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1801 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1861 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1921 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1981 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2041 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2101 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2161 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2221 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2521 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3001 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3061 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3121 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3181 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     3241 gtgcactctc agtacaatct gctctgatgc cgcatagtta agcca
//
