LOCUS       sequence-457075-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology repair plasmid for endogenous tagging of AP1muA at the 
            C-terminus with SNAP tag and a 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 6633)
  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 6633)
  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: AP1muA-SNAP-V5-polyA-Puro HR
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(706..730)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(830..1621,1622..1690))
                     /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     1453..1472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1691..1795)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1894..1916
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1908..1925
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1909..1925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3119..3661
                     /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="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     CDS             3662..3703
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3830..3951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3980..4000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3985..4314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4165..4300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4407..5006
                     /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(4407..4426)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4903..4923
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5236..5357
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6446..6462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6446..6462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6459..6481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt caagtcagcg
      661 taaatgggta gggggcttca aatcgtcctc gtgataccaa ttcggagcct gcttttttgt
      721 acaaacttgt tgataatggc aattcaagga tcttcaccta gatcctttta aattaaaaat
      781 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      841 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      901 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
      961 tgataccgcg agagccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1021 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1081 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1141 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1201 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1261 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1321 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1381 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1441 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1501 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1561 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1621 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1681 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1741 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1801 ttccccgaaa agtgccagat acctgaaaca aaacccatcg tacggccaag gaagtctcca
     1861 ataactgtga tccaccacaa gcgccagggt tttcccagtc acgacgttgt aaaacgacgg
     1921 ccagtcatgc ataatccgca cgcatctgga ataaggaagt gccattccgc ctgaccttgg
     1981 ggcagcacag ggccgagctt tggctcccca gcacaggggc tcatgctccc ctccctggac
     2041 tggggtgagt ggtcagtctc tgcccatcgc cactagggcc agagctgttg gctaagctgg
     2101 ggcagggcct aggcctgagc gcctcccccg tctgctcccc agggcggcaa ggagtacctg
     2161 atgcgggccc acttcggcct gcctagtgtg gaggccgaag acaaggaggg caagcccccg
     2221 atcagtgtca agttcgagat cccttacttc actacctccg gcatccaggt acgtaaggcc
     2281 caggcggccg gggcccagaa cagggacaga ggccgcaggt gacagatatc aatgcagcca
     2341 gcttgggaaa gggtgtcagt cagctgcaat caggaccctg ctgtgctgtg ggcctctgcc
     2401 tccccacaga cagggtgagg ccaggctgcc tctgctcagc cctgacccct gagctcaaag
     2461 ctcactcatt ctgccccaaa ctagcatttg aggctctgcc gtccactcat gccccatggg
     2521 acctcttttc cctggcaccc cctcaaaagc ccacagaggg aggagaggct cccagagagc
     2581 caggggcctt ctggccgggc cccccaagca ggctgctgag gccgtgtctg caagccacat
     2641 cctggctgct caccacttcc agaggcggct ctgccctccc acacatttgc atggtccaga
     2701 caaatccctt tcacccccct gaggcctgtg ctcatcctgt cgtcatggcc tcccctgcca
     2761 gccccaggct gccccagcag gaaagattaa ggggggacca acagggcggg agcctgcggt
     2821 gctcagggcc ctgctacctg tgccccaggt gcgctacctg aagatcattg agaagagtgg
     2881 gtaccaggcc ctgccctggg tgcgttatat cacgcagaat ggaggtgagt gaggccctac
     2941 ccgtggggtg gggttgtact gaggggtcct ctgtggctgc ccccagggtg gagccatcgg
     3001 gtcgggtccc gaaagcaggg agggtgcaga gcccagcatg acgcctccct cctgtctcct
     3061 agattaccag ctccggaccc agggatcagg gtcaggctct ggttccggct caggatccga
     3121 caaagactgc gaaatgaagc gcaccaccct ggatagccct ctgggcaagc tggaactgtc
     3181 tgggtgcgaa cagggcctgc accgtatcat cttcctgggc aaaggaacat ctgccgccga
     3241 cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga ccagagccac tgatgcaggc
     3301 caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt tccctgtgcc
     3361 agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc tgtggaaact
     3421 gctgaaagtg gtgaagttcg gagaggtcat cagctacagc cacctggccg ccctggccgg
     3481 caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc ggaaatcccg tgcccattct
     3541 gatcccctgc caccgggtgg tgcagggcga cctggacgtg gggggctacg agggcgggct
     3601 cgccgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc ctgggctggg
     3661 tggcaagccc atccccaacc ccctgctggg cctggacagc acctgagcta gcccgcgact
     3721 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3781 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3841 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3901 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3961 ttgtaaggcg gccgcctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4201 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4261 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaacctt
     4321 gcatgcctgc aggtcggccg ccacgaccgg tgccgccacc atcccctgac ccacgcccct
     4381 gacccctcac aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca
     4441 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4501 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4561 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4621 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4681 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     4741 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     4801 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     4861 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     4921 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     4981 gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa
     5041 ggagcgcacg accccatggc tccgaccgaa gccacccggg gcggccccgc cgaccccgca
     5101 cccgcccccg aggcccaccg actctagagg atcataatca gccataccac atttgtagag
     5161 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5221 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5281 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5341 ctcatcaatg tatcttatca tgtctggaat tcggggctgt cgcagccaac accccagcct
     5401 cggagctcct ggtggcagca ccaggggaca cacctgccaa acccaccaga tggaggggcc
     5461 ctccctggtc tctggccacc ctcccagcct ctgcccaggg acccctgcct tccccaggcc
     5521 atctgctctg ccgtcgacac tcgtctcaga agcccctttc ccagaagagg ctggtcttca
     5581 agaagtctcg tttctttgcc cctgaagtca gtttcagggg aaggatgtga aatttttccg
     5641 tgtagaggtt acagcctttt atgctgttga gctcccaggt accaaaaagc ttggccaacg
     5701 cttgccagcc agccagctgc aggtggcatc tgccacgaag gaagcgccag cctcgccagg
     5761 ccagcagggg cgtcgttttg ttgccatttt gttgaacgtt atgggtttat gggtgttcct
     5821 ggaacttgtc tttgtgcatt cgttgctgtt tgtgttaccc tcactgtccc catgtcccac
     5881 ccacgtccta cggcactcag gaagcacttg gtgaggacga gccctcaccc ttcttgtctt
     5941 ccttcccggc agcgcccgca gcgggccatt tacacgtcga ggctggcacc tggcgcgctc
     6001 gggggccact gtagcgtctg cctgctccct ggactcgcag gcctcgcctg tggcgccttc
     6061 ccagggccag cctgggtcac gagatgctgt cactcagcca gatcagtatt gacccaccag
     6121 gggaggtggg gtttggtgag agacgccagc ctcagacttt ttcccactga gggtccagag
     6181 agcggggcca cgtgtcaccc acgtctgcgc ttggtcaccc gtcctcccca ccctgtgtgt
     6241 gtttatgtca tagttacatt aaattccatt cattgaatac tccacagcct tcctctgatg
     6301 cagcgtggtc ccaactcctc ctgcgtgacc acagggcttg cctggttttg tcgccgacct
     6361 tccctgtggc cccacgaggt gtcaccagca ggctaggtgg aggctcagtg atgataagtc
     6421 tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6481 tcagagggca caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc
     6541 agttcggcgt atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6601 caggaaccgt aaaaaggccg cgttgctggc gtt
//
