LOCUS       sequence-171205-        5691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Rab12 Q101L, constitutively GTP-bound 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5691)
  AUTHORS   Xu J, Fotouhi M, McPherson PS
  TITLE     Phosphorylation of the exchange factor DENND3 by ULK in response to 
            starvation activates Rab12 and induces autophagy.
  JOURNAL   EMBO Rep. 2015 Jun;16(6):709-18. doi: 10.15252/embr.201440006. Epub 
            2015 Apr 29.
  PUBMED    25925668
REFERENCE   2  (bases 1 to 5691)
  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"; doi: 
            "10.15252/embr.201440006"; journalName: "EMBO Rep"; date: "2015-06";
            volume: "16"; issue: "6"; pages: "709-18"
COMMENT     Sequence Label: Flag Rab12 Q101L
FEATURES             Location/Qualifiers
     source          1..5691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..36)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(20..36)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(58..74)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(59..75)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(130..149)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(131..149)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    423..544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(460..479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     514..533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(551..1006)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(688..709)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     900..919
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1033..1137
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1162..1182)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1206..1416
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1228..1424
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1275..1410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1459..2253
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1459..2253
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1513..1532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2123..2142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2441..2460)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2485..2532
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2861..3449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3350..3369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3624..3927
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3928..4131
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4082..4102
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4177..4195
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      4230..4239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4239..4262
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 gctctagatt gcggccgcgg tcatagctgt ttcctgaaca gatcccatca agcttatcga
       61 taccgtcgac ctcgaggggg ggcccggtac cttaattaat taaggtacca ggtaagtgta
      121 cccaattcgc cctatagtga gtcgtattac aattcactcg atcgcccttc ccaacagttg
      181 cgcagcctga atggcgaatg gagatccaat ttttaagtgt ataatgtgtt aaactactga
      241 ttctaattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg cccctcagtc
      301 ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
      361 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
      421 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      541 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      661 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      721 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      781 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      841 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      901 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      961 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     1021 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1081 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatcct
     1141 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1201 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1261 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1321 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     1381 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     1441 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     1501 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     1561 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     1621 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     1681 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     1741 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     1801 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     1861 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1921 caggatgatc tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1981 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2041 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2101 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc
     2161 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2221 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     2281 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     2341 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     2401 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     2461 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     2521 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2581 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     2641 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     2701 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2761 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2821 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2881 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2941 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3001 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3061 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3481 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3541 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     3601 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3661 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3721 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3781 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3841 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3901 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3961 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     4021 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     4081 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     4141 aaccgtcaga tccgctagcg attacgccaa gctcgaaatt aaccctcact aaagggaaca
     4201 aaagctggag ctccaccgcg gtggcggccg ccaccatgga ttacaaggat gacgacgata
     4261 aggcccgggc ggatccgggc gccgcgctgc agaggcgggc cgggggcggc ggcggtctgg
     4321 gcgcgggctc cccggcgctg tcgggcggcc agggccgccg gaggaagcag ccccccaggc
     4381 cggccgactt caagttgcag gtcatcatta tcggctcccg cggcgtgggc aagaccagcc
     4441 tgatggagcg cttcaccgac gacaccttct gcgaggcctg caagtccacc gtgggtgttg
     4501 acttcaaaat caaaactgta gagctaagag gaaagaaaat tagattacag atctgggaca
     4561 cagcaggtct ggagagattc aacagcatta cctcagctta ttacagaagt gccaagggga
     4621 tcatattagt atatgatatc actaagaagg agacatttga tgatttgccg aaatggatga
     4681 agatgattga taagtatgct tcagaagatg cagagcttct cttagttgga aataagttgg
     4741 actgtgaaac ggacagagaa atcaccaggc agcaggggga aaagtttgca cagcagatca
     4801 ctgggatgcg gttctgtgaa gcaagtgcca aggataactt caatgtggac gagatatttt
     4861 tgaaacttgt cgatgacatt ctgaaaaaga tgcctctgga tattttaagg aatgagttgt
     4921 ccaatagtat cctgtcgtta caaccagagc ctgagatacc gccagaactg cctccaccaa
     4981 gaccacatgt ccgatgctgt tgatttccta ctttggagac aaagtggaaa tgattcctgg
     5041 aaaggggaaa aaacgttcta ttctgcacta caatcatttt gacaatttcc tttcgcactt
     5101 tgtaatccaa gtcagagcta tacactaact tgtaaatatg catatatgca atcctgggta
     5161 agttttggtt ataagttacc tatttccctc caaattatta tatttcattc attaccccag
     5221 tgtctagtgt acatacactg ggaaacctag tacttctaat atgaagaatg ggagaaatga
     5281 aaggtataat gtttcttgaa ataaataata taattgtcct tattaattat attatgagga
     5341 cagaagatat tctgataaga gagaacgtgg tgctttgctt accgttttaa agaaaatttg
     5401 taaaactaaa gactttttga aaaaaagcta tcttaagtgc tttttcttta tttacaagac
     5461 atttccccca gtggtagcat ctgaagtatt ggagtgtttc tgccacgaag caaagctcca
     5521 ttcatggccg tcatggaagg ttatttatta atgttacata atggtagaat attactagtt
     5581 agagggttgg atttgacttg gtcctaaggc cacagaatct ctctcatggc ttcctaaggg
     5641 atgtaccttt atgcttttaa gaactacaaa gattcaataa agaaagaaat g
//
