LOCUS       sequence-456965-        6957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology repair plasmid for endogenous tagging of AP1muA at the 
            C-terminus with monomeric StayGold. Contains a hygromycin resistance
            cassette for selection of edited cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6957)
  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 6957)
  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-mStayGold-polyA-Hygro HR
FEATURES             Location/Qualifiers
     source          1..6957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..132
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    181..302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(429..1106)
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     primer_bind     complement(2300..2317)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2300..2316)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2309..2331)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        2430..2534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2535..2603,2604..3395)
                     /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(2753..2772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    3495..3519
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      3636..4224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4125..4144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4377..4399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4396..4412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4396..4412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5492..5613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5743..6768)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     enhancer        join(6921..6957,1..155)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
       61 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
      121 gacacacatt ccacagctgg ttctttccgc ctcagggcgg ccgccttaca atttacgcct
      181 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      241 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      301 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      361 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcta gagtcgcggg
      421 ctagcttaca ggtgggcctc cagggtctcg ctctggatga tgtggtctct ctcctcggtg
      481 tcgtccttgc tctgggtgta ctgctttctg atccagtgga atggcacatc gggggctggc
      541 tgattgtgca ggggcttgat gatggtgttc tggtaggcct ccacgcactt gctctcgtcg
      601 ctcagcagag ggtactgcag ggtcactgtg cactccacgc cgtcatctat ggggatgtgc
      661 tgcacctcgt tgggcaggct cacgtccatg tcgccggtca gcacggggct gttctccttg
      721 aagtcctggc cggtgaactc cacgatgttg tggtaggtgc cgttcttcat gaagtgctgg
      781 tgcttggcgt ggatgctgcc gtcgcccttg tactggatgt gtctgtcgta ggtgaagccc
      841 tcgggcatca cctcgtggaa ccagttcttc aggccgctgg ggtacttggt gtagtacttc
      901 atgccatagc cgaagctagt tcccagggcg gcccaagaca ttggcagttt gccgctggtg
      961 cacacgtact tgcctttgtg gctgccctcg tggctattgc cctcgccctc gccttccacg
     1021 gtgaagctct tgccgttgat ggtgcccttc agctggaact tgaagggcac cacgccggta
     1081 aacagctcct cgcctgtaga caccatggat cctgagccgg aaccagagcc tgaccctgat
     1141 ccctgggtcc ggagctggta atctaggaga caggagggag gcgtcatgct gggctctgca
     1201 ccctccctgc tttcgggacc cgacccgatg gctccaccct gggggcagcc acagaggacc
     1261 cctcagtaca accccacccc acgggtaggg cctcactcac ctccattctg cgtgatataa
     1321 cgcacccagg gcagggcctg gtacccactc ttctcaatga tcttcaggta gcgcacctgg
     1381 ggcacaggta gcagggccct gagcaccgca ggctcccgcc ctgttggtcc ccccttaatc
     1441 tttcctgctg gggcagcctg gggctggcag gggaggccat gacgacagga tgagcacagg
     1501 cctcaggggg gtgaaaggga tttgtctgga ccatgcaaat gtgtgggagg gcagagccgc
     1561 ctctggaagt ggtgagcagc caggatgtgg cttgcagaca cggcctcagc agcctgcttg
     1621 gggggcccgg ccagaaggcc cctggctctc tgggagcctc tcctccctct gtgggctttt
     1681 gagggggtgc cagggaaaag aggtcccatg gggcatgagt ggacggcaga gcctcaaatg
     1741 ctagtttggg gcagaatgag tgagctttga gctcaggggt cagggctgag cagaggcagc
     1801 ctggcctcac cctgtctgtg gggaggcaga ggcccacagc acagcagggt cctgattgca
     1861 gctgactgac accctttccc aagctggctg cattgatatc tgtcacctgc ggcctctgtc
     1921 cctgttctgg gccccggccg cctgggcctt acgtacctgg atgccggagg tagtgaagta
     1981 agggatctcg aacttgacac tgatcggggg cttgccctcc ttgtcttcgg cctccacact
     2041 aggcaggccg aagtgggccc gcatcaggta ctccttgccg ccctggggag cagacggggg
     2101 aggcgctcag gcctaggccc tgccccagct tagccaacag ctctggccct agtggcgatg
     2161 ggcagagact gaccactcac cccagtccag ggaggggagc atgagcccct gtgctgggga
     2221 gccaaagctc ggccctgtgc tgccccaagg tcaggcggaa tggcacttcc ttattccaga
     2281 tgcgtgcgga ttatgcatga ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2341 gcgcttgtgg tggatcacag ttattggaga cttccttggc cgtacgatgg gttttgtttc
     2401 aggtatctgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2461 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2521 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2581 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2641 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2701 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2761 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2821 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2881 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2941 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3001 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3061 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3121 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3181 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3241 gtgagcgtgg ctctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3301 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3361 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3421 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttg
     3481 aattgccatt atcaacaagt ttgtacaaaa aagcaggctc cgaattggta tcacgaggac
     3541 gatttgaagc cccctaccca tttacgctga cttgacggga cggcggcttt gttgaataga
     3601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4261 ttttgctggc cttttgctca catgttcttt ccagcgacat atgccatacg ccgaactgaa
     4321 ctccacctaa tgtcttggag attgtcggat agcgcggaat aggattgtgc cctctgagcg
     4381 gataacaatt tcacacagga aacagctatg accatgacac atgcatccac catcgcagac
     4441 ttatcatcac tgagcctcca cctagcctgc tggtgacacc tcgtggggcc acagggaagg
     4501 tcggcgacaa aaccaggcaa gccctgtggt cacgcaggag gagttgggac cacgctgcat
     4561 cagaggaagg ctgtggagta ttcaatgaat ggaatttaat gtaactatga cataaacaca
     4621 cacagggtgg ggaggacggg tgaccaagcg cagacgtggg tgacacgtgg ccccgctctc
     4681 tggaccctca gtgggaaaaa gtctgaggct ggcgtctctc accaaacccc acctcccctg
     4741 gtgggtcaat actgatctgg ctgagtgaca gcatctcgtg acccaggctg gccctgggaa
     4801 ggcgccacag gcgaggcctg cgagtccagg gagcaggcag acgctacagt ggcccccgag
     4861 cgcgccaggt gccagcctcg acgtgtaaat ggcccgctgc gggcgctgcc gggaaggaag
     4921 acaagaaggg tgagggctcg tcctcaccaa gtgcttcctg agtgccgtag gacgtgggtg
     4981 ggacatgggg acagtgaggg taacacaaac agcaacgaat gcacaaagac aagttccagg
     5041 aacacccata aacccataac gttcaacaaa atggcaacaa aacgacgccc ctgctggcct
     5101 ggcgaggctg gcgcttcctt cgtggcagat gccacctgca gctggctggc tggcaagcgt
     5161 tggccaagct ttttggtacc tgggagctca acagcataaa aggctgtaac ctctacacgg
     5221 aaaaatttca catccttccc ctgaaactga cttcaggggc aaagaaacga gacttcttga
     5281 agaccagcct cttctgggaa aggggcttct gagacgagtg tcgacggcag agcagatggc
     5341 ctggggaagg caggggtccc tgggcagagg ctgggagggt ggccagagac cagggagggc
     5401 ccctccatct ggtgggtttg gcaggtgtgt cccctggtgc tgccaccagg agctccgagg
     5461 ctggggtgtt ggctgcgaca gccccgaatt ctaagataca ttgatgagtt tggacaaacc
     5521 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5581 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
     5641 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     5701 ttcatagaag gcggcggtgg aatcgaaatc tcgtagtacg tgctattcct ttgccctcgg
     5761 acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca
     5821 gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac
     5881 gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct
     5941 ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccgcg gcgatcctgc
     6001 aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg
     6061 cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca
     6121 gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc
     6181 gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc
     6241 ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg
     6301 atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc
     6361 gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatcgcat ccatggcctc
     6421 cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc
     6481 ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aactccccaa tgtcaagcac
     6541 ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa
     6601 accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa
     6661 agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc
     6721 gatcagaaac ttctcgacag acgtcgcggt gagttcaggc tttttcatat cgatctttgc
     6781 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     6841 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gttaggggcg
     6901 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgcttt
//
