LOCUS       sequence-457129-        5874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology repair plasmid for endogenous tagging of Rab11 at the 
            N-terminus with SNAP tag and a 3x V5 epitope tag. Contains a 
            puromycin resistance cassette for selection of edited cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5874)
  AUTHORS   Stockhammer A, Adarska P, Natalia V, Heuhsen A, Klemt A, Bregu G, 
            Harel S, Rodilla-Ramirez C, Spalt C, Ozsoy E, Leupold P, Grindel A, 
            Fox E, Mejedo JO, Zehtabian A, Ewers H, Puchkov D, Haucke V, 
            Bottanelli F
  TITLE     ARF1 compartments direct cargo flow via maturation into recycling 
            endosomes.
  JOURNAL   Nat Cell Biol. 2024 Oct 4. doi: 10.1038/s41556-024-01518-4.
  PUBMED    39367144
REFERENCE   2  (bases 1 to 5874)
  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: "Nat Cell 
            Biol. 2024 Oct 4. doi: 10.1038/s41556-024-01518-4."
COMMENT     Sequence Label: loxP-Puro-loxP-3xV5-SNAP-Rab11 HR
FEATURES             Location/Qualifiers
     source          1..5874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    264..385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(301..320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     355..374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(395..428)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             459..500
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             504..545
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             549..590
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             600..1145
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(1872..1889)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1944..1960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1944..1960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1957..1979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2132..2720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2212..2231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2837..2861)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2961..3752,3753..3821))
                     /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     3584..3603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3822..3926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4025..4047
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4039..4056
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4040..4056
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4823..4856)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4882..4902)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4887..5216
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5067..5202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5309..5874,1..34)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5309..5328)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5805..5825
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
       61 gaccgaaagg agcgcacgac cccatggctc cgaccgaagc cacccggggc ggccccgccg
      121 accccgcacc cgcccccgag gcccaccgac tctagaggat cataatcagc cataccacat
      181 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
      241 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
      301 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
      361 tgtccaaact catcaatgta tcttatcatg tctgataact tcgtatagca tacattatac
      421 gaagttatcc tcgaggtatc atctaacgga tcggtatggg caagcccatc cccaaccccc
      481 tgctgggcct ggacagcacc ggcggtaaac caattccaaa tccattgttg ggtttggatt
      541 ctactggcgg taaaccgatt ccgaatccgc tgctgggtct ggatagcacc ggctccggaa
      601 tggacaaaga ctgcgaaatg aagcgcacca ccctggatag ccctctgggc aagctggaac
      661 tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct gggcaaagga acatctgccg
      721 ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg cggaccagag ccactgatgc
      781 aggccaccgc ctggctcaac gcctactttc accagcctga ggccatcgag gagttccctg
      841 tgccagccct gcaccaccca gtgttccagc aggagagctt tacccgccag gtgctgtgga
      901 aactgctgaa agtggtgaag ttcggagagg tcatcagcta cagccacctg gccgccctgg
      961 ccggcaatcc cgccgccacc gccgccgtga aaaccgccct gagcggaaat cccgtgccca
     1021 ttctgatccc ctgccaccgg gtggtgcagg gcgacctgga cgtggggggc tacgagggcg
     1081 gtctcgccgt gaaagagtgg ctgctggccc acgagggcca cagactgggc aagcctgggc
     1141 tgggtggatc cggatcaggg tcaggctctg gttccggctc aatgggcacc cgcgacgacg
     1201 aatatgacta cctctttaaa ggtgaggcca tgggctctcg cactctacac agtcctcgtt
     1261 cggggacccg ggccactccc ggtggaccct cgtgccggcc acccctgcac tgatataggc
     1321 ctccctcagc ccttcctttt tgtgcggttc cgtctcctac ccagctcagc ctcttctccc
     1381 ccgctcagac aggggtcccc atcacatgcc gctctctgag cgacctctcc ataggccttc
     1441 gctggcctca gagcccctcc ctgcgtgtcc ttcccctggc ggactgcctt ctcccacatc
     1501 gtcgaattcc tttccccggg ttctacggcc ccgccgctcc tcccaccatc tctcttttcg
     1561 ggtgtagcgc cccctccccc tcggcgtaca cccttcccag ctcgcgtcct ctcccgaagc
     1621 ccctctgacg ggttcttcgc ttccctcttg gccttgcctt cggtgcagac tcccattaca
     1681 ggtctttttc ttatctgccc tctttccccc ctcgagcgag cgtctcttct gtatctcccc
     1741 tccacttgga cggccgcccc tctaagcttc tcccatcccg ggtccaggac cccgtaaaat
     1801 ctgagcgggc caagggcctc cgcccccacc ctccgccccg cggttctcgc ggtccttaac
     1861 cttctctgac cgctccctct cggtaccagg ctaggtggag gctcagtgat gataagtctg
     1921 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1981 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
     2041 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
     2101 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2161 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2221 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2281 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2341 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2401 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2461 acgacttatc gccactggca gcagctactg gtaacaggat tagcagagcg aggtatgtag
     2521 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2581 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2641 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2701 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
     2761 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtcct
     2821 cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg caattcaagg
     2881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3061 gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg ctcaccggct
     3121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3841 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3901 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga tacctgaaac
     3961 aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca agcgccaggg
     4021 ttttcccagt cacgacgttg taaaacgacg gccagtcatg cataatccgc acgcatctgg
     4081 aataaggaag tgccattccg cctgacctgc ggccgccccc ccgccaggta gcagatagca
     4141 cctgaaatgc agtgggaaat ttgtgaggtc ttcataggca aggattaaat tttacatctc
     4201 catcttaaga tgtgtcatag ttgttcaatt acttagtgac caaaaaagtg aattaaagat
     4261 gttgtcaaga ttaaaaaaac aaaaaccaat gcaggttcca ccgagatttg aactcggatc
     4321 gctggattca gagtccagag tgctaaccat tacaccatgg aaccgccacg catgtgtagc
     4381 tgccttcggc tgtctaatcc tcagagaacc ccgcccccat ccacaaaccc accactcaca
     4441 ggcggtcccg cctggttcca gcgagccgct tccggcacgg tagctcgaga aatgagcaag
     4501 cggccactaa gactatggta gctaggagtt ccaggactca gtttcccctt tgagcctcct
     4561 ttagcgacta aagcttgaag ccccacgcat ctcgactctc gcgcacaccg cccttgttgg
     4621 gctcaggggc ggggcgccgc ccccggaagt acttcccctt aaaggctggg gcctgccgga
     4681 aatggcgcag cggcagggag gggctcttca cccagtccgg cagttgaagc tcggcgctcg
     4741 ggttacccct gcagcgacgc cccctggtcc cacagatacc actgctgctc ccgccctttc
     4801 gctcctcggc agcgcagcta gcataacttc gtatagcata cattatacga agttatgatg
     4861 ccgcatagtt aagccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4921 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4981 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5041 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5101 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5161 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5221 ttgcatgcct gcaggtcggc cgccacgacc ggtgccgcca ccatcccctg acccacgccc
     5281 ctgacccctc acaaggagac gaccttccat gaccgagtac aagcccacgg tgcgcctcgc
     5341 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5401 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5461 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5521 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5581 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5641 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5701 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5761 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5821 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcg
//
