LOCUS       sequence-178001-        5618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A bacterial expression plasmid encoding GDP locked Rab8a 
            (Rab8a-T22N) with N-terminus GST..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5618)
  AUTHORS   Madugula V, Lu L
  TITLE     A ternary complex comprising transportin1, Rab8 and the ciliary 
            targeting signal directs proteins to ciliary membranes.
  JOURNAL   J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15.
  PUBMED    27633000
REFERENCE   2  (bases 1 to 5618)
  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: "J Cell 
            Sci."; date: "2016-10-15"; volume: "129(20)"; pages: "3922-3934. 
            Epub 2016 Sep 15"
COMMENT     Sequence Label: GST-Rab8a-TN
FEATURES             Location/Qualifiers
     source          1..5618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     494..513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     747..764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        837..914
                     /gene="lacI"
                     /label=lacI promoter
     CDS             915..1997
                     /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(934..953)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2046..2051,2052..2069,2070..2076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2084..2100
                     /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     2108..2124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2108..2124
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2120..2647
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2140..2157)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2140..2156)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2149..2171)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2749..2754,2755..2770,2771..2777)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2785..2801
                     /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)."
     CDS             2824..3477
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3435..3457
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3484..3501
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4234..4256)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4401..4419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4487..4591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4592..4660,4661..5452)
                     /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(4810..4829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
       61 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      121 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
      181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      781 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca
      841 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
      901 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
      961 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     1021 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     1081 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     1141 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     1201 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     1261 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     1321 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     1381 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     1441 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     1501 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     1561 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     1621 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     1681 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     1741 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     1801 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     1861 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     1921 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     1981 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccc
     2041 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2101 tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg
     2161 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2221 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2281 cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg
     2341 gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca
     2401 cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca atccgccgtt
     2461 tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     2521 gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaattagct tatcgactgc
     2581 acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag
     2641 gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt
     2701 tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat
     2761 catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggaaacagta
     2821 ttcatgtccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt
     2881 cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat
     2941 aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt
     3001 gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac
     3061 aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt
     3121 ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa
     3181 gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat
     3241 aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt
     3301 gatgttgttt tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt
     3361 aaaaaacgta ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata
     3421 gcatggcctt tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg
     3481 gatctggttc cgcgtggatc agaattcgcg aagacctacg attacctgtt caagctgctg
     3541 ctgatcgggg actcgggggt ggggaagaac tgtgtcctgt tccgcttctc cgaggacgcc
     3601 ttcaactcca cttttatctc caccatagga attgacttta aaattaggac catagagctc
     3661 gatggcaaga gaattaaact gcagatatgg gacacagccg gtcaggaacg gtttcggacg
     3721 atcacaacgg cctactacag gggtgcaatg ggcatcatgc tggtctacga catcaccaac
     3781 gagaagtcct tcgacaacat ccggaactgg attcgcaaca ttgaggagca cgcctctgca
     3841 gacgtcgaaa agatgatact cgggaacaag tgtgatgtga atgacaagag acaagtttcc
     3901 aaggaacggg gagaaaagct ggccctcgac tatggaatca agttcatgga gaccagcgcg
     3961 aaggccaaca tcaatgtgga aaatgcattt ttcactctcg ccagagatat caaagcaaaa
     4021 atggacaaaa aattggaagg caacagcccc caggggagca accagggagt caaaatcaca
     4081 ccggaccagc agaagaggag cagctttttc cgatgtgttc ttctgtaggg atccttaaca
     4141 attctagact ccatgggtcg actcgagctc aagcttaatt catcgtgact gactgacgat
     4201 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4261 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4321 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     4381 taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4801 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     4861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5101 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     5161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5581 ccactgagcg tcagaccccg tagaaaagat caaaggat
//
