LOCUS       sequence-455371-        5337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein purification of Rab1A (S20C).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5337)
  AUTHORS   Morstein J, Bowcut V, Fernando M, Yang Y, Zhu L, Jenkins ML, Evans 
            JT, Guiley KZ, Peacock DM, Krahnke S, Lin Z, Taran KA, Huang BJ, 
            Stephen AG, Burke JE, Lightstone FC, Shokat KM
  TITLE     Targeting Ras-, Rho-, and Rab-family GTPases via a conserved cryptic
            pocket.
  JOURNAL   Cell. 2024 Sep 6:S0092-8674(24)00908-5. doi: 
            10.1016/j.cell.2024.08.017.
  PUBMED    39255801
REFERENCE   2  (bases 1 to 5337)
  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: "Cell. 2024 
            Sep 6:S0092-8674(24)00908-5. doi: 10.1016/j.cell.2024.08.017."
COMMENT     Sequence Label: pProEx Rab1A(S20C) CL
FEATURES             Location/Qualifiers
     source          1..5337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    229..245
                     /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)."
     primer_bind     234..256
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             278..295
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             317..337
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      849..858
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1026..1043)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1196..1282
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1374..1401
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1420..1511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1512..1580,1581..2372)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1730..1749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2543..3131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3032..3051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3285..3302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3379..3807)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3489..3510)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3701..3720
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3845..3864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4037..4059
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4137..4214
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4215..5297
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4234..4253)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tttgacagct tatcatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
       61 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
      121 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
      181 gaaatgagct gttgacaatt aatcatccgg tccgtataat ctgtggaatt gtgagcggat
      241 aacaatttca cacaggaaac agaccatgtc gtactaccat caccatcacc atcacgatta
      301 cgatatccca acgaccgaaa acctgtattt tcagggcgcc atgggatcca tgagttcaat
      361 gaatcccgaa tatgattact tatttaaact cctgctgatt ggcgactgcg gtgttggcaa
      421 gtcgtccctg ttgctgcgtt tcgcggacga cacctatact gagtcctaca tcagcaccat
      481 cggcgtcgac ttcaagatcc gtacgattga actggatggt aaaaccatta aattgcagat
      541 ttgggatacc gctggccaag agcgctttcg taccatcacc agcagttact accgcggtgc
      601 gcacggcatc atcgtggttt atgatgttac cgaccaggag agcttcaaca acgtgaaaca
      661 gtggctgcaa gagatcgatc gttatgcgag cgaaaacgtt aacaagttgt tggtcggcaa
      721 taagtctgat ctgaccacca aaaaagttgt ggactacacg actgcgaaag aatttgcgga
      781 cagcctgggt attccgtttc tggaaacgtc tgccaagaac gcaaccaatg tggaacaaag
      841 cttcatgacc atggcggcag agattaagaa gagaatgggt ccgggtgcca cggctggtta
      901 atgagaattc aaaggcctac gtcgacgagc tcactagtcg cggccgcttt cgaatctaga
      961 gcctgcagtc tcgaggcatg cggtaccaag cttggctgtt ttggcggatg agagaagatt
     1021 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     1081 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     1141 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     1201 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1261 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     1321 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     1381 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     1441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1501 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1561 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1621 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1681 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1741 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1801 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1861 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1921 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1981 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2041 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2101 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2161 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2221 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2281 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2341 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2401 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2461 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2521 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2581 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2641 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2701 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2761 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2821 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2881 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2941 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3001 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3061 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3301 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3361 accgcataat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3421 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3481 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3541 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3601 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3661 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3721 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3781 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcta tggtgcactc
     3841 tcagtacaat ctgctctgat gccgcatagt taagccagta ccagtcacgt agcgatatcg
     3901 ctacgtgact ggtacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3961 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4021 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4081 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttgaca
     4141 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     4201 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     4261 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     4321 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     4381 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     4441 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     4501 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     4561 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     4621 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     4681 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     4741 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     4801 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     4861 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     4921 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     4981 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     5041 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     5101 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     5161 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcaccc aatacgcaaa
     5221 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5281 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt gatctgg
//
