LOCUS       sequence-382136-        4777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged Rab8 in mammalian cells and zebrafish embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4777)
  AUTHORS   Aljiboury AA, Ingram E, Krishnan N, Ononiwu F, Pal D, Manikas J, 
            Taveras C, Hall NA, Da Silva J, Freshour J, Hehnly H
  TITLE     Rab8, Rab11, and Rab35 coordinate lumen and cilia formation during 
            zebrafish left-right organizer development.
  JOURNAL   PLoS Genet. 2023 May 15;19(5):e1010765. doi: 
            10.1371/journal.pgen.1010765. eCollection 2023 May.
  PUBMED    37186603
REFERENCE   2  (bases 1 to 4777)
  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: "PLoS 
            Genet."; date: "2023-05-15"; volume: "19(5):e1010765. doi"; pages: "
            10.1371/journal.pgen.1010765. eCollection 2023 May"
COMMENT     Sequence Label: pCS2-FLAG-Rab8
FEATURES             Location/Qualifiers
     source          1..4777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             109..132
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    829..963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(853..872)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     907..926
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1076..1096)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1076..1094)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1115..1131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1115..1131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1128..1150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1139..1155
                     /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(1163..1169,1170..1187,1188..1193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1208..1229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1346..1363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1517..2105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1597..1616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2276..3067,3068..3136))
                     /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     2899..2918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3137..3241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3267..3722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3616..3635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3793..4777
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagc cgctagaaga cccaagctgg ctagcatgga ctacaaagac
      121 gatgacgaca agctcaatcg agctcaagct tcgaattctg cgaagaccta cgattacctg
      181 ttcaagctgc tgctgatcgg ggactcgggg gtggggaaga cctgtgtcct gttccgcttc
      241 tccgaggacg ccttcaactc aactttcatc tccaccatag gaattgactt taaaattagg
      301 accatagagc tcgatggcaa gagaattaag ctacagatat gggacacagc tggtcaagaa
      361 cggtttcgga cgatcacaac agcctattac aggggcgcaa tgggcatcat gctggtctat
      421 gacatcacca acgagaaatc ctttgacaat atccggaact ggattcggaa cattgaggag
      481 catgcttctg cagatgtcga aaagatgata ctcggaaaca agtgtgatgt gaacgacaaa
      541 agacaagttt ccaaggaacg gggagaaaag ctggccctgg actatggaat caagttcatg
      601 gagaccagtg cgaaggccaa catcaatgtg gagaacgcat ttttcactct cgccagagac
      661 atcaaagcaa agatggacaa aaaattggaa ggcaacagtc cccaagggag caaccaggga
      721 gtcaaaatca caccagacca gcagaagagg agcagctttt tccgatgtgt tcttctgtga
      781 tcgccagcac agtctgcagt ctagaactat agtgagtcgt attacgtaga tccagacatg
      841 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
      901 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
      961 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1021 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     1081 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1141 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     1201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3241 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     3301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3721 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3781 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
     3841 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
     3901 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
     3961 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
     4021 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
     4081 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
     4141 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
     4201 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
     4261 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
     4321 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
     4381 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
     4441 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
     4501 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
     4561 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
     4621 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
     4681 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
     4741 atgggaggtc tatataagca atgctcgttt agggaac
//
