LOCUS       sequence-357407-        4880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor with homologous arms for Rab11b to knock in SFFV-T2A-Puromycin
            resistance-PolyA cassette (for knock out).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4880)
  AUTHORS   Gemperle J, Harrison TS, Flett C, Adamson AD, Caswell PT
  TITLE     On demand expression control of endogenous genes with DExCon, 
            DExogron and LUXon reveals differential dynamics of Rab11 family 
            members.
  JOURNAL   Elife. 2022 Jun 16;11. pii: 76651. doi: 10.7554/eLife.76651.
  PUBMED    35708998
REFERENCE   2  (bases 1 to 4880)
  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: "Elife."; 
            date: "2022-06-16"; volume: "11. pii: 76651. doi"; pages: " 
            10.7554/eLife.76651"
COMMENT     Sequence Label: pJET-62-SFFV-T2A-PuroR-polyA-R11b
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             15..68
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(709..726)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    715..939
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(join(1507..1513,1514..1531,1532..1537))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1552..1573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1693..1710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1864..2452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1944..1963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2623..3414,3415..3483))
                     /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     3246..3265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3484..3588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3656..3674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3980..3998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4048..4066
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4389..4796
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 ctagtggcag cggagagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
       61 ccggccctag gctcgagatg acggaatata agcccacagt cagattggct acgagagatg
      121 atgttccgcg agccgtgcgg acgctcgcag ccgcttttgc agactatcct gctacgcgac
      181 atactgttga ccctgatagg catatcgaaa gagtaaccga acttcaggag ctctttctga
      241 ctagggtcgg tttggatatt gggaaggtct gggtagccga cgacggcgcc gctgtcgccg
      301 tatggacgac tccagagagc gtagaagcgg gagcggtatt tgctgagatc ggtccacgga
      361 tggcagaatt gtccggcagt agacttgcgg ctcaacagca aatggaagga cttctcgcac
      421 cgcatcgacc caaagagccc gcctggtttc ttgctacggt aggggtgtcc cccgaccacc
      481 agggtaaggg tctcggaagt gctgttgtcc ttccaggagt ggaagcggca gagcgcgcgg
      541 gtgttcctgc gtttctggag acctcagccc caaggaacct tcctttttat gaacgcttgg
      601 gatttactgt cacggccgat gttgaggttc ctgaaggacc tcggacatgg tgcatgacaa
      661 ggaaaccagg ggcgtgatct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
      721 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
      781 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
      841 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
      901 acaatagcag gcatgctggg gatgcggtgg gctctatggg tcgacggaac cggtgctgga
      961 agtggtaccc gggacgacga gtacgactac ctattcaaag gtgcggccgg tggggcacag
     1021 acgggcgaag tcgtggcggc gaggcggcgg gcagacgggc caaggccgcg ctatctttct
     1081 agaagatctc ctacaatatt ctcagctgcc atggaaaatc gatgttcttc ttttattctc
     1141 tcaagatttt caggctgtat attaaaactt atattaagaa ctatgctaac cacctcatca
     1201 ggaaccgttg taggtggcgt gggttttctt ggcaatcgac tctcatgaaa actacgagct
     1261 aaatattcaa tatgttcctc ttgaccaact ttattctgca ttttttttga acgaggttta
     1321 gagcaagctt caggaaactg agacaggaat tttattaaaa atttaaattt tgaagaaagt
     1381 tcagggttaa tagcatccat tttttgcttt gcaagttcct cagcattctt aacaaaagac
     1441 gtctcttttg acatgtttaa agtttaaacc tcctgtgtga aattattatc cgctcataat
     1501 tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1561 ctaactcaca ttaattgcgt tgcgctcact gccaattgct ttccagtcgg gaaacctgtc
     1621 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2281 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3601 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3661 gcgtatcacg aggccgcccc tgcagccgaa ttatattatt tttgccaaat aatttttaac
     3721 aaaagctctg aagtcttctt catttaaatt cttagatgat acttcatctg gaaaattgtc
     3781 ccaattagta gcatcacgct gtgagtaagt tctaaaccat ttttttattg ttgtattatc
     3841 tctaatctta ctactcgatg agttttcggt attatctcta tttttaactt ggagcaggtt
     3901 ccattcattg tttttttcat catagtgaat aaaatcaact gctttaacac ttgtgcctga
     3961 acaccatatc catccggcgt aatacgactc actataggga gagcggccgc cagatcttcc
     4021 ggatggctcg agtttttcag caagatataa tacgactcac tatagggcag tcttccgccc
     4081 cgccctcttg agctcgcccc tgataggctg gcgcgtccgt cacttcaaaa aggtccgcat
     4141 tccttccgcc tttctccagg acaccgaggg cgaggagggt ggtaccaagc ggcgcccacc
     4201 ctcagagcac tacttccatc tctgattggc ttcgctgggt gcccgtcgct actccactcg
     4261 ccgctgatag gccattcaac cgccgctctg cgcggcgccg gctccgcccc cgtcgggtgt
     4321 ttgtggtggg gctgcggagt cgccgatccc gccggaagcg ccaggacaat gggatcgggt
     4381 gggactaggt aacgccattt tgcaaggcat ggaaaaatac caaaccaaga atagagaagt
     4441 tcagatcaag ggcgggtaca tgaaaatagc taacgttggg ccaaacagga tatctgcggt
     4501 gagcagtttc ggccccggcc cggggccaag aacagatggt caccgcagtt tcggccccgg
     4561 cccgaggcca agaacagatg gtccccagat atggcccaac cctcagcagt ttcttaagac
     4621 ccatcagatg tttccaggct cccccaagga cctgaaatga ccctgcgcct tatttgaatt
     4681 aaccaatcag cctgcttctc gcttctgttc gcgcgcttct gcttcccgag ctctataaaa
     4741 gagctcacaa cccctcactc ggcgcgccag tcctccgaca gactgagtcg cccggggggg
     4801 atctggagct ctcgagaatt ctcacgcgtc tgcaggatat caagcttgcg gtaccgcggg
     4861 cccgggatcc atggtgagca
//
