LOCUS       sequence-492078-        5796 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  mammalian expression of Rab12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5796)
  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 5796)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 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 wild type
FEATURES             Location/Qualifiers
     source          1..5796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        658..676
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      711..720
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             720..743
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2225..2241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2225..2241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2263..2279)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2264..2280)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2335..2354)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2336..2354)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2628..2749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2665..2684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2719..2738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2756..3211)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2893..2914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3105..3124
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3238..3342
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3344..3701
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3344..3535
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3367..3387)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3552..3687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3736..4530
                     /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             3736..4530
                     /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(3790..3809)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4400..4419
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4718..4737)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4762..4809
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5138..5726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5627..5646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctggtttagt gaaccgtcag atccgctagc gattacgcca agctcgaaat
      661 taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc gccaccatgg
      721 attacaagga tgacgacgat aaggcccggg cggatccggg cgccgcgctg cagaggcggg
      781 ccgggggcgg cggcggtctg ggcgcgggct ccccggcgct gtcgggcggc cagggccgcc
      841 ggaggaagca gccccccagg ccggccgact tcaagttgca ggtcatcatt atcggctccc
      901 gcggcgtggg caagaccagc ctgatggagc gcttcaccga cgacaccttc tgcgaggcct
      961 gcaagtccac cgtgggtgtt gacttcaaaa tcaaaactgt agagctaaga ggaaagaaaa
     1021 ttagattaca gatctgggac acagcaggtc aggagagatt caacagcatt acctcagctt
     1081 attacagaag tgccaagggg atcatattag tatatgatat cactaagaag gagacatttg
     1141 atgatttgcc gaaatggatg aagatgattg ataagtatgc ttcagaagat gcagagcttc
     1201 tcttagttgg aaataagttg gactgtgaaa cggacagaga aatcaccagg cagcaggggg
     1261 aaaagtttgc acagcagatc actgggatgc ggttctgtga agcaagtgcc aaggataact
     1321 tcaatgtgga cgagatattt ttgaaacttg tcgatgacat tctgaaaaag atgcctctgg
     1381 atattttaag gaatgagttg tccaatagta tcctgtcgtt acaaccagag cctgagatac
     1441 cgccagaact gcctccacca agaccacatg tccgatgctg ttgatttcct actttggaga
     1501 caaagtggaa atgattcctg gaaaggggaa aaaacgttct attctgcact acaatcattt
     1561 tgacaatttc ctttcgcact ttgtaatcca agtcagagct atacactaac ttgtaaatat
     1621 gcatatatgc aatcctgggt aagttttggt tataagttac ctatttccct ccaaattatt
     1681 atatttcatt cattacccca gtgtctagtg tacatacact gggaaaccta gtacttctaa
     1741 tatgaagaat gggagaaatg aaaggtataa tgtttcttga aataaataat ataattgtcc
     1801 ttattaatta tattatgagg acagaagata ttctgataag agagaacgtg gtgctttgct
     1861 taccgtttta aagaaaattt gtaaaactaa agactttttg aaaaaaagct atcttaagtg
     1921 ctttttcttt atttacaaga catttccccc agtggtagca tctgaagtat tggagtgttt
     1981 ctgccacgaa gcaaagctcc attcatggcc gtcatggaag gttatttatt aatgttacat
     2041 aatggtagaa tattactagt tagagggttg gatttgactt ggtcctaagg ccacagaatc
     2101 tctctcatgg cttcctaagg gatgtacctt tatgctttta agaactacaa agattcaata
     2161 aagaaagaaa tgtttttgaa aaaaaaaaaa aaaaaaaggg cggccgctct agattgcggc
     2221 cgcggtcata gctgtttcct gaacagatcc catcaagctt atcgataccg tcgacctcga
     2281 gggggggccc ggtaccttaa ttaattaagg taccaggtaa gtgtacccaa ttcgccctat
     2341 agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag cctgaatggc
     2401 gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     2461 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     2521 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2581 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2641 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2701 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2761 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2821 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2881 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2941 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3001 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3061 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3121 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3181 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3241 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3301 taaccctgat aaatgcttca ataatattga aaaaggaaga atcctgaggc ggaaagaacc
     3361 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3481 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3541 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3601 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3661 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3721 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3781 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3841 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3901 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3961 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4021 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4081 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4141 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4201 gatgatctgg acgaagaaca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4261 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4321 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4381 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagaact tggcggcgaa
     4441 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4501 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4561 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4621 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4681 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4741 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4801 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4861 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4921 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4981 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5041 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5101 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5161 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5221 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5281 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5341 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5401 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5461 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5521 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5581 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5641 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5701 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5761 ccttttgctg gccttttgct cacatgttct ttcctg
//
