LOCUS       sequence-456981-        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"
     enhancer        867..1058
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1015..1035
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     protein_bind    1060..1093
                     /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(1860..1877)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1860..1876)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1869..1891)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1990..2094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2095..2163,2164..2955)
                     /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(2313..2332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    3055..3079
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      3196..3784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3685..3704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3937..3959
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3956..3972
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3956..3972
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4027..4044
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4771..5445)
                     /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(5446..5472)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    5482..5515
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5516..5637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5527..5546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5581..5600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5767..6078,1..714))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
       61 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
      121 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
      181 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
      241 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
      301 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat
      361 ggcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
      421 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc
      481 aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt
      541 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
      601 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
      661 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tcatatcgat
      721 ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
      781 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagtta
      841 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
      901 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
      961 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     1021 actgacacac attccacagc tggttctttc cgcctcagga taacttcgta taatgtatgc
     1081 tatacgaagt tatgctagct gcgctgccga ggagcgaaag ggcgggagca gcagtggtat
     1141 ctgtgggacc agggggcgtc gctgcagggg taacccgagc gccgagcttc aactgccgga
     1201 ctgggtgaag agcccctccc tgccgctgcg ccatttccgg caggccccag cctttaaggg
     1261 gaagtacttc cgggggcggc gccccgcccc tgagcccaac aagggcggtg tgcgcgagag
     1321 tcgagatgcg tggggcttca agctttagtc gctaaaggag gctcaaaggg gaaactgagt
     1381 cctggaactc ctagctacca tagtcttagt ggccgcttgc tcatttctcg agctaccgtg
     1441 ccggaagcgg ctcgctggaa ccaggcggga ccgcctgtga gtggtgggtt tgtggatggg
     1501 ggcggggttc tctgaggatt agacagccga aggcagctac acatgcgtgg cggttccatg
     1561 gtgtaatggt tagcactctg gactctgaat ccagcgatcc gagttcaaat ctcggtggaa
     1621 cctgcattgg tttttgtttt tttaatcttg acaacatctt taattcactt ttttggtcac
     1681 taagtaattg aacaactatg acacatctta agatggagat gtaaaattta atccttgcct
     1741 atgaagacct cacaaatttc ccactgcatt tcaggtgcta tctgctacct ggcggggggg
     1801 cggccgcagg tcaggcggaa tggcacttcc ttattccaga tgcgtgcgga ttatgcatga
     1861 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgcttgtgg tggatcacag
     1921 ttattggaga cttccttggc cgtacgatgg gttttgtttc aggtatctgg cacttttcgg
     1981 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2101 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2161 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2221 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2281 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2341 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2401 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2461 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2521 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2581 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2641 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2701 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2761 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ctctcgcggt
     2821 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2881 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2941 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3001 cttcattttt aatttaaaag gatctaggtg aagatccttg aattgccatt atcaacaagt
     3061 ttgtacaaaa aagcaggctc cgaattggta tcacgaggac gatttgaagc cccctaccca
     3121 tttacgctga cttgacggga cggcggcttt gttgaataga gcgtcagacc ccgtagaaaa
     3181 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3241 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3301 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3361 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3421 gttaccagta gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3661 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3721 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3781 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3841 catgttcttt ccagcgacat atgccatacg ccgaactgaa ctccacctaa tgtcttggag
     3901 attgtcggat agcgcggaat aggattgtgc cctctgagcg gataacaatt tcacacagga
     3961 aacagctatg accatgacac atgcatccac catcgcagac ttatcatcac tgagcctcca
     4021 cctagcctgg taccgagagg gagcggtcag agaaggttaa ggaccgcgag aaccgcgggg
     4081 cggagggtgg gggcggaggc ccttggcccg ctcagatttt acggggtcct ggacccggga
     4141 tgggagaagc ttagaggggc ggccgtccaa gtggagggga gatacagaag agacgctcgc
     4201 tcgagggggg aaagagggca gataagaaaa agacctgtaa tgggagtctg caccgaaggc
     4261 aaggccaaga gggaagcgaa gaacccgtca gaggggcttc gggagaggac gcgagctggg
     4321 aagggtgtac gccgaggggg agggggcgct acacccgaaa agagagatgg tgggaggagc
     4381 ggcggggccg tagaacccgg ggaaaggaat tcgacgatgt gggagaaggc agtccgccag
     4441 gggaaggaca cgcagggagg ggctctgagg ccagcgaagg cctatggaga ggtcgctcag
     4501 agagcggcat gtgatgggga cccctgtctg agcgggggag aagaggctga gctgggtagg
     4561 agacggaacc gcacaaaaag gaagggctga gggaggccta tatcagtgca ggggtggccg
     4621 gcacgagggt ccaccgggag tggcccgggt ccccgaacga ggactgtgta gagtgcgaga
     4681 gcccatggcc tcacctttaa agaggtagtc atattcgtcg tcgcgggtgc ccattgagcc
     4741 ggaaccagag cctgaccctg atccggatcc caggtgggcc tccagggtct cgctctggat
     4801 gatgtggtct ctctcctcgg tgtcgtcctt gctctgggtg tactgctttc tgatccagtg
     4861 gaatggcaca tcgggggctg gctgattgtg caggggcttg atgatggtgt tctggtaggc
     4921 ctccacgcac ttgctctcgt cgctcagcag agggtactgc agggtcactg tgcactccac
     4981 gccgtcatct atggggatgt gctgcacctc gttgggcagg ctcacgtcca tgtcgccggt
     5041 cagcacgggg ctgttctcct tgaagtcctg gccggtgaac tccacgatgt tgtggtaggt
     5101 gccgttcttc atgaagtgct ggtgcttggc gtggatgctg ccgtcgccct tgtactggat
     5161 gtgtctgtcg taggtgaagc cctcgggcat cacctcgtgg aaccagttct tcaggccgct
     5221 ggggtacttg gtgtagtact tcatgccata gccgaagcta gttcccaggg cggcccaaga
     5281 cattggcagt ttgccgctgg tgcacacgta cttgcctttg tggctgccct cgtggctatt
     5341 gccctcgccc tcgccttcca cggtgaagct cttgccgttg atggtgccct tcagctggaa
     5401 cttgaagggc accacgccgg taaacagctc ctcgcctgta gacacagcgt aatctggaac
     5461 atcgtatggg tacatctcga gataacttcg tataatgtat gctatacgaa gttattaaga
     5521 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     5581 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttggg
     5641 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     5701 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtag
     5761 tacgtgctat tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
     5821 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
     5881 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
     5941 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
     6001 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
     6061 ctgctccata caagccaa
//
