LOCUS       sequence-197130-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Rab8-DN (T22N) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  AUTHORS   Burke MC, Li FQ, Cyge B, Arashiro T, Brechbuhl HM, Chen X, Siller 
            SS, Weiss MA, O'Connell CB, Love D, Westlake CJ, Reynolds SD, 
            Kuriyama R, Takemaru K
  TITLE     Chibby promotes ciliary vesicle formation and basal body docking 
            during airway cell differentiation.
  JOURNAL   J Cell Biol. 2014 Oct 13;207(1):123-37. doi: 10.1083/jcb.201406140.
  PUBMED    25313408
REFERENCE   2  (bases 1 to 4743)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2014-10-13- 13"; volume: "207"; 
            issue: "1"; pages: "123-37"
COMMENT     Sequence Label: HA-Rab8a-DN (T22N)
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1070..1096
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1749..1768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1750..1768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1773..1907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1797..1816)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1851..1870
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2020..2040)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2020..2038)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2059..2075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2059..2075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2072..2094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2083..2099
                     /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)."
     promoter        complement(join(2107..2113,2114..2131,2132..2137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2152..2173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2290..2307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2461..3049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2541..2560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3220..4011,4012..4080))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3843..3862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4081..4185)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4211..4666)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4348..4369)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4560..4579
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat accatgcgat acccatatga
     1081 tgtgccagat tacgctggat ccatggcgaa gacctacgat tacctgttca agctgctgct
     1141 gatcggggac tcgggggtgg ggaagaactg tgtcctgttc cgcttctccg aggacgcctt
     1201 caactccact tttatctcca ccataggaat tgactttaaa attaggacca tagagctcga
     1261 tggcaagaga attaaactgc agatatggga cacagccggt caggaacggt ttcggacgat
     1321 cacaacggcc tactacaggg gtgcaatggg catcatgctg gtctacgaca tcaccaacga
     1381 gaagtccttc gacaacatcc ggaactggat tcgcaacatt gaggagcacg cctctgcaga
     1441 cgtcgaaaag atgatactcg ggaacaagtg tgatgtgaat gacaagagac aagtttccaa
     1501 ggaacgggga gaaaagctgg ccctcgacta tggaatcaag ttcatggaga ccagcgcgaa
     1561 ggccaacatc aatgtggaaa atgcattttt cactctcgcc agagatatca aagcaaaaat
     1621 ggacaaaaaa ttggaaggca acagccccca ggggagcaac cagggagtca aaatcacacc
     1681 ggaccagcag aagaggagca gctttttccg atgtgttctt ctgtagctcg agcctctaga
     1741 ttctgcagcc ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
     1801 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1861 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1921 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
     1981 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     2041 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2101 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4201 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4261 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4321 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4381 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4441 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4501 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4561 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4621 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     4681 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
     4741 acc
//
