LOCUS       sequence-456961-        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"
     CDS             1079..1621
                     /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             1622..1663
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1790..1911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1940..1960)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1945..2274
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2125..2260
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2367..2966
                     /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(2367..2386)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2863..2883
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3196..3317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4406..4422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4406..4422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4419..4441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(4594..5182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4674..4693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(5299..5323)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(5423..6214,6215..6283))
                     /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     6046..6065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6284..6388)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6487..6509
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6501..6518
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6502..6518
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggggtgagt ggtcagtctc tgcccatcgc cactagggcc agagctgttg gctaagctgg
       61 ggcagggcct aggcctgagc gcctcccccg tctgctcccc agggcggcaa ggagtacctg
      121 atgcgggccc acttcggcct gcctagtgtg gaggccgaag acaaggaggg caagcccccg
      181 atcagtgtca agttcgagat cccttacttc actacctccg gcatccaggt acgtaaggcc
      241 caggcggccg gggcccagaa cagggacaga ggccgcaggt gacagatatc aatgcagcca
      301 gcttgggaaa gggtgtcagt cagctgcaat caggaccctg ctgtgctgtg ggcctctgcc
      361 tccccacaga cagggtgagg ccaggctgcc tctgctcagc cctgacccct gagctcaaag
      421 ctcactcatt ctgccccaaa ctagcatttg aggctctgcc gtccactcat gccccatggg
      481 acctcttttc cctggcaccc cctcaaaagc ccacagaggg aggagaggct cccagagagc
      541 caggggcctt ctggccgggc cccccaagca ggctgctgag gccgtgtctg caagccacat
      601 cctggctgct caccacttcc agaggcggct ctgccctccc acacatttgc atggtccaga
      661 caaatccctt tcacccccct gaggcctgtg ctcatcctgt cgtcatggcc tcccctgcca
      721 gccccaggct gccccagcag gaaagattaa ggggggacca acagggcggg agcctgcggt
      781 gctcagggcc ctgctacctg tgccccaggt gcgctacctg aagatcattg agaagagtgg
      841 gtaccaggcc ctgccctggg tgcgttatat cacgcagaat ggaggtgagt gaggccctac
      901 ccgtggggtg gggttgtact gaggggtcct ctgtggctgc ccccagggtg gagccatcgg
      961 gtcgggtccc gaaagcaggg agggtgcaga gcccagcatg acgcctccct cctgtctcct
     1021 agattaccag ctccggaccc agggatcagg gtcaggctct ggttccggct caggatccga
     1081 caaagactgc gaaatgaagc gcaccaccct ggatagccct ctgggcaagc tggaactgtc
     1141 tgggtgcgaa cagggcctgc accgtatcat cttcctgggc aaaggaacat ctgccgccga
     1201 cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga ccagagccac tgatgcaggc
     1261 caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt tccctgtgcc
     1321 agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc tgtggaaact
     1381 gctgaaagtg gtgaagttcg gagaggtcat cagctacagc cacctggccg ccctggccgg
     1441 caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc ggaaatcccg tgcccattct
     1501 gatcccctgc caccgggtgg tgcagggcga cctggacgtg gggggctacg agggcgggct
     1561 cgccgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc ctgggctggg
     1621 tggcaagccc atccccaacc ccctgctggg cctggacagc acctgagcta gcccgcgact
     1681 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1741 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1801 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1861 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     1921 ttgtaaggcg gccgcctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1981 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2041 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2101 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2161 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2221 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaacctt
     2281 gcatgcctgc aggtcggccg ccacgaccgg tgccgccacc atcccctgac ccacgcccct
     2341 gacccctcac aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca
     2401 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     2461 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     2521 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     2581 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     2641 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     2701 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     2761 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     2821 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     2881 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     2941 gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa
     3001 ggagcgcacg accccatggc tccgaccgaa gccacccggg gcggccccgc cgaccccgca
     3061 cccgcccccg aggcccaccg actctagagg atcataatca gccataccac atttgtagag
     3121 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3181 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3241 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3301 ctcatcaatg tatcttatca tgtctggaat tcggggctgt cgcagccaac accccagcct
     3361 cggagctcct ggtggcagca ccaggggaca cacctgccaa acccaccaga tggaggggcc
     3421 ctccctggtc tctggccacc ctcccagcct ctgcccaggg acccctgcct tccccaggcc
     3481 atctgctctg ccgtcgacac tcgtctcaga agcccctttc ccagaagagg ctggtcttca
     3541 agaagtctcg tttctttgcc cctgaagtca gtttcagggg aaggatgtga aatttttccg
     3601 tgtagaggtt acagcctttt atgctgttga gctcccaggt accaaaaagc ttggccaacg
     3661 cttgccagcc agccagctgc aggtggcatc tgccacgaag gaagcgccag cctcgccagg
     3721 ccagcagggg cgtcgttttg ttgccatttt gttgaacgtt atgggtttat gggtgttcct
     3781 ggaacttgtc tttgtgcatt cgttgctgtt tgtgttaccc tcactgtccc catgtcccac
     3841 ccacgtccta cggcactcag gaagcacttg gtgaggacga gccctcaccc ttcttgtctt
     3901 ccttcccggc agcgcccgca gcgggccatt tacacgtcga ggctggcacc tggcgcgctc
     3961 gggggccact gtagcgtctg cctgctccct ggactcgcag gcctcgcctg tggcgccttc
     4021 ccagggccag cctgggtcac gagatgctgt cactcagcca gatcagtatt gacccaccag
     4081 gggaggtggg gtttggtgag agacgccagc ctcagacttt ttcccactga gggtccagag
     4141 agcggggcca cgtgtcaccc acgtctgcgc ttggtcaccc gtcctcccca ccctgtgtgt
     4201 gtttatgtca tagttacatt aaattccatt cattgaatac tccacagcct tcctctgatg
     4261 cagcgtggtc ccaactcctc ctgcgtgacc acagggcttg cctggttttg tcgccgacct
     4321 tccctgtggc cccacgaggt gtcaccagca ggctaggtgg aggctcagtg atgataagtc
     4381 tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4441 tcagagggca caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc
     4501 agttcggcgt atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctct
     5221 attcaacaaa gccgccgtcc cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc
     5281 ctcgtgatac caattcggag cctgcttttt tgtacaaact tgttgataat ggcaattcaa
     5341 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5401 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5461 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5521 gggagggctt accatctggc cccagtgctg caatgatacc gcgagagcca cgctcaccgg
     5581 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5641 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5701 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5761 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5821 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5881 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5941 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6001 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6061 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6121 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6181 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6241 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6301 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6361 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca gatacctgaa
     6421 acaaaaccca tcgtacggcc aaggaagtct ccaataactg tgatccacca caagcgccag
     6481 ggttttccca gtcacgacgt tgtaaaacga cggccagtca tgcataatcc gcacgcatct
     6541 ggaataagga agtgccattc cgcctgacct tggggcagca cagggccgag ctttggctcc
     6601 ccagcacagg ggctcatgct cccctccctg gac
//
