LOCUS       sequence-456964-        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"
     primer_bind     486..505
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(598..927)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      612..747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     912..932
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    958..991
                     /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(1758..1775)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1758..1774)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1767..1789)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1888..1992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1993..2061,2062..2853)
                     /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     complement(2211..2230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    2953..2977
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      3094..3682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3583..3602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3835..3857
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3854..3870
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3854..3870
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3925..3942
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4669..5214)
                     /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(5224..5265)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5269..5310)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5314..5355)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    5386..5419
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5429..5550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5440..5459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5494..5513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5780..5874,1..505))
                     /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(join(5863..5874,1..9))
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct
       61 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc
      121 cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca
      181 tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg
      241 cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt
      301 cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca
      361 gctcggtgac ccgctcgatg tggcggtccg ggtcgacggt gtggcgcgtg gcggggtagt
      421 cggcgaacgc ggcggcgagg gtgcgtacgg cccgggggac gtcgtcgcgg gtggcgaggc
      481 gcaccgtggg cttgtactcg gtcatggaag gtcgtctcct tgtgaggggt caggggcgtg
      541 ggtcagggga tggtggcggc accggtcgtg gcggccgacc tgcaggcatg caagcttttt
      601 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
      661 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
      721 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
      781 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      841 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      901 ggactttcca caccctaact gacacacatt ccacagctgg cttaactatg cggcatcata
      961 acttcgtata atgtatgcta tacgaagtta tgctagctgc gctgccgagg agcgaaaggg
     1021 cgggagcagc agtggtatct gtgggaccag ggggcgtcgc tgcaggggta acccgagcgc
     1081 cgagcttcaa ctgccggact gggtgaagag cccctccctg ccgctgcgcc atttccggca
     1141 ggccccagcc tttaagggga agtacttccg ggggcggcgc cccgcccctg agcccaacaa
     1201 gggcggtgtg cgcgagagtc gagatgcgtg gggcttcaag ctttagtcgc taaaggaggc
     1261 tcaaagggga aactgagtcc tggaactcct agctaccata gtcttagtgg ccgcttgctc
     1321 atttctcgag ctaccgtgcc ggaagcggct cgctggaacc aggcgggacc gcctgtgagt
     1381 ggtgggtttg tggatggggg cggggttctc tgaggattag acagccgaag gcagctacac
     1441 atgcgtggcg gttccatggt gtaatggtta gcactctgga ctctgaatcc agcgatccga
     1501 gttcaaatct cggtggaacc tgcattggtt tttgtttttt taatcttgac aacatcttta
     1561 attcactttt ttggtcacta agtaattgaa caactatgac acatcttaag atggagatgt
     1621 aaaatttaat ccttgcctat gaagacctca caaatttccc actgcatttc aggtgctatc
     1681 tgctacctgg cgggggggcg gccgcaggtc aggcggaatg gcacttcctt attccagatg
     1741 cgtgcggatt atgcatgact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1801 gcttgtggtg gatcacagtt attggagact tccttggccg tacgatgggt tttgtttcag
     1861 gtatctggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1981 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2041 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2101 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2161 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2221 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2281 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2341 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2401 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2461 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2521 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2581 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2641 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2701 gagcgtggct ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2761 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2821 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2881 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttgaa
     2941 ttgccattat caacaagttt gtacaaaaaa gcaggctccg aattggtatc acgaggacga
     3001 tttgaagccc cctacccatt tacgctgact tgacgggacg gcggctttgt tgaatagagc
     3061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3181 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3241 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3301 cctcgctctg ctaatcctgt taccagtagc tgctgccagt ggcgataagt cgtgtcttac
     3361 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3421 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3481 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3541 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3601 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3661 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3721 ttgctggcct tttgctcaca tgttctttcc agcgacatat gccatacgcc gaactgaact
     3781 ccacctaatg tcttggagat tgtcggatag cgcggaatag gattgtgccc tctgagcgga
     3841 taacaatttc acacaggaaa cagctatgac catgacacat gcatccacca tcgcagactt
     3901 atcatcactg agcctccacc tagcctggta ccgagaggga gcggtcagag aaggttaagg
     3961 accgcgagaa ccgcggggcg gagggtgggg gcggaggccc ttggcccgct cagattttac
     4021 ggggtcctgg acccgggatg ggagaagctt agaggggcgg ccgtccaagt ggaggggaga
     4081 tacagaagag acgctcgctc gaggggggaa agagggcaga taagaaaaag acctgtaatg
     4141 ggagtctgca ccgaaggcaa ggccaagagg gaagcgaaga acccgtcaga ggggcttcgg
     4201 gagaggacgc gagctgggaa gggtgtacgc cgagggggag ggggcgctac acccgaaaag
     4261 agagatggtg ggaggagcgg cggggccgta gaacccgggg aaaggaattc gacgatgtgg
     4321 gagaaggcag tccgccaggg gaaggacacg cagggagggg ctctgaggcc agcgaaggcc
     4381 tatggagagg tcgctcagag agcggcatgt gatggggacc cctgtctgag cgggggagaa
     4441 gaggctgagc tgggtaggag acggaaccgc acaaaaagga agggctgagg gaggcctata
     4501 tcagtgcagg ggtggccggc acgagggtcc accgggagtg gcccgggtcc ccgaacgagg
     4561 actgtgtaga gtgcgagagc ccatggcctc acctttaaag aggtagtcat attcgtcgtc
     4621 gcgggtgccc attgagccgg aaccagagcc tgaccctgat ccggatccac ccagcccagg
     4681 cttgcccagt ctgtggccct cgtgggccag cagccactct ttcacggcga gaccgccctc
     4741 gtagcccccc acgtccaggt cgccctgcac cacccggtgg caggggatca gaatgggcac
     4801 gggatttccg ctcagggcgg ttttcacggc ggcggtggcg gcgggattgc cggccagggc
     4861 ggccaggtgg ctgtagctga tgacctctcc gaacttcacc actttcagca gtttccacag
     4921 cacctggcgg gtaaagctct cctgctggaa cactgggtgg tgcagggctg gcacagggaa
     4981 ctcctcgatg gcctcaggct ggtgaaagta ggcgttgagc caggcggtgg cctgcatcag
     5041 tggctctggt ccgcccagca cggcggctgg ggcaggcact tccacggcgt cggcggcaga
     5101 tgttcctttg cccaggaaga tgatacggtg caggccctgt tcgcacccag acagttccag
     5161 cttgcccaga gggctatcca gggtggtgcg cttcatttcg cagtctttgt ccattccgga
     5221 gccggtgcta tccagaccca gcagcggatt cggaatcggt ttaccgccag tagaatccaa
     5281 acccaacaat ggatttggaa ttggtttacc gccggtgctg tccaggccca gcagggggtt
     5341 ggggatgggc ttgcccatac cgatccgtta gatgatacct cgaggataac ttcgtataat
     5401 gtatgctata cgaagttatc agacatgata agatacattg atgagtttgg acaaaccaca
     5461 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     5521 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     5581 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     5641 atggctgatt atgatcctct agagtcggtg ggcctcgggg gcgggtgcgg ggtcggcggg
     5701 gccgccccgg gtggcttcgg tcggagccat ggggtcgtgc gctcctttcg gtcgggcgct
     5761 gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     5821 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aagg
//
