LOCUS       sequence-457125-        6078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology repair plasmid for endogenous tagging of Rab11 at the 
            N-terminus with monomeric StayGold and an HA epitope tag. 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 6078)
  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 6078)
  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-Hygro-loxP-HA-mStayGold-Rab11 HR
FEATURES             Location/Qualifiers
     source          1..6078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     742..764
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     761..777
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     761..777
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             832..849
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1576..2250)
                     /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="VSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGK
                     YVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSI
                     HAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTLQ
                     YPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETLE
                     AHL"
     CDS             complement(2251..2277)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2287..2320
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2321..2442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2332..2351)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2386..2405
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2572..3597)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     enhancer        3750..3941
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3898..3918
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    3943..3976
                     /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(4743..4760)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4743..4759)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4752..4774)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        4873..4977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4978..5046,5047..5838)
                     /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(5196..5215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    5938..5962
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtagctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttccagc
      661 gacatatgcc atacgccgaa ctgaactcca cctaatgtct tggagattgt cggatagcgc
      721 ggaataggat tgtgccctct gagcggataa caatttcaca caggaaacag ctatgaccat
      781 gacacatgca tccaccatcg cagacttatc atcactgagc ctccacctag cctggtaccg
      841 agagggagcg gtcagagaag gttaaggacc gcgagaaccg cggggcggag ggtgggggcg
      901 gaggcccttg gcccgctcag attttacggg gtcctggacc cgggatggga gaagcttaga
      961 ggggcggccg tccaagtgga ggggagatac agaagagacg ctcgctcgag gggggaaaga
     1021 gggcagataa gaaaaagacc tgtaatggga gtctgcaccg aaggcaaggc caagagggaa
     1081 gcgaagaacc cgtcagaggg gcttcgggag aggacgcgag ctgggaaggg tgtacgccga
     1141 gggggagggg gcgctacacc cgaaaagaga gatggtggga ggagcggcgg ggccgtagaa
     1201 cccggggaaa ggaattcgac gatgtgggag aaggcagtcc gccaggggaa ggacacgcag
     1261 ggaggggctc tgaggccagc gaaggcctat ggagaggtcg ctcagagagc ggcatgtgat
     1321 ggggacccct gtctgagcgg gggagaagag gctgagctgg gtaggagacg gaaccgcaca
     1381 aaaaggaagg gctgagggag gcctatatca gtgcaggggt ggccggcacg agggtccacc
     1441 gggagtggcc cgggtccccg aacgaggact gtgtagagtg cgagagccca tggcctcacc
     1501 tttaaagagg tagtcatatt cgtcgtcgcg ggtgcccatt gagccggaac cagagcctga
     1561 ccctgatccg gatcccaggt gggcctccag ggtctcgctc tggatgatgt ggtctctctc
     1621 ctcggtgtcg tccttgctct gggtgtactg ctttctgatc cagtggaatg gcacatcggg
     1681 ggctggctga ttgtgcaggg gcttgatgat ggtgttctgg taggcctcca cgcacttgct
     1741 ctcgtcgctc agcagagggt actgcagggt cactgtgcac tccacgccgt catctatggg
     1801 gatgtgctgc acctcgttgg gcaggctcac gtccatgtcg ccggtcagca cggggctgtt
     1861 ctccttgaag tcctggccgg tgaactccac gatgttgtgg taggtgccgt tcttcatgaa
     1921 gtgctggtgc ttggcgtgga tgctgccgtc gcccttgtac tggatgtgtc tgtcgtaggt
     1981 gaagccctcg ggcatcacct cgtggaacca gttcttcagg ccgctggggt acttggtgta
     2041 gtacttcatg ccatagccga agctagttcc cagggcggcc caagacattg gcagtttgcc
     2101 gctggtgcac acgtacttgc ctttgtggct gccctcgtgg ctattgccct cgccctcgcc
     2161 ttccacggtg aagctcttgc cgttgatggt gcccttcagc tggaacttga agggcaccac
     2221 gccggtaaac agctcctcgc ctgtagacac agcgtaatct ggaacatcgt atgggtacat
     2281 ctcgagataa cttcgtataa tgtatgctat acgaagttat taagatacat tgatgagttt
     2341 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2401 attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc
     2461 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     2521 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagtacgt gctattcctt
     2581 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
     2641 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
     2701 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
     2761 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg
     2821 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
     2881 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
     2941 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
     3001 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
     3061 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
     3121 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
     3181 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
     3241 catggcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa
     3301 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga actccccaat
     3361 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
     3421 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
     3481 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
     3541 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc
     3601 gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     3661 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     3721 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     3781 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     3841 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     3901 ctaactgaca cacattccac agctggttct ttccgcctca ggataacttc gtataatgta
     3961 tgctatacga agttatgcta gctgcgctgc cgaggagcga aagggcggga gcagcagtgg
     4021 tatctgtggg accagggggc gtcgctgcag gggtaacccg agcgccgagc ttcaactgcc
     4081 ggactgggtg aagagcccct ccctgccgct gcgccatttc cggcaggccc cagcctttaa
     4141 ggggaagtac ttccgggggc ggcgccccgc ccctgagccc aacaagggcg gtgtgcgcga
     4201 gagtcgagat gcgtggggct tcaagcttta gtcgctaaag gaggctcaaa ggggaaactg
     4261 agtcctggaa ctcctagcta ccatagtctt agtggccgct tgctcatttc tcgagctacc
     4321 gtgccggaag cggctcgctg gaaccaggcg ggaccgcctg tgagtggtgg gtttgtggat
     4381 gggggcgggg ttctctgagg attagacagc cgaaggcagc tacacatgcg tggcggttcc
     4441 atggtgtaat ggttagcact ctggactctg aatccagcga tccgagttca aatctcggtg
     4501 gaacctgcat tggtttttgt ttttttaatc ttgacaacat ctttaattca cttttttggt
     4561 cactaagtaa ttgaacaact atgacacatc ttaagatgga gatgtaaaat ttaatccttg
     4621 cctatgaaga cctcacaaat ttcccactgc atttcaggtg ctatctgcta cctggcgggg
     4681 gggcggccgc aggtcaggcg gaatggcact tccttattcc agatgcgtgc ggattatgca
     4741 tgactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgcttg tggtggatca
     4801 cagttattgg agacttcctt ggccgtacga tgggttttgt ttcaggtatc tggcactttt
     4861 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4921 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4981 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5041 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5101 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5161 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5221 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5281 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5341 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5401 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5461 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5521 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5581 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5641 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc
     5701 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5761 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5821 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5881 aaacttcatt tttaatttaa aaggatctag gtgaagatcc ttgaattgcc attatcaaca
     5941 agtttgtaca aaaaagcagg ctccgaattg gtatcacgag gacgatttga agccccctac
     6001 ccatttacgc tgacttgacg ggacggcggc tttgttgaat agagcgtcag accccgtaga
     6061 aaagatcaaa ggatcttc
//
