LOCUS       sequence-254821-        5277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HDR tempalte for tagging of endogenous human RAB18 N-terminus with 
            sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  AUTHORS   Salo VT, Li S, Vihinen H, Holtta-Vuori M, Szkalisity A, Horvath P, 
            Belevich I, Peranen J, Thiele C, Somerharju P, Zhao H, Santinho A, 
            Thiam AR, Jokitalo E, Ikonen E
  TITLE     Seipin Facilitates Triglyceride Flow to Lipid Droplet and 
            Counteracts Droplet Ripening via Endoplasmic Reticulum Contact.
  JOURNAL   Dev Cell. 2019 Jun 6. pii: S1534-5807(19)30388-0. doi: 
            10.1016/j.devcel.2019.05.016.
  PUBMED    31178403
REFERENCE   2  (bases 1 to 5277)
  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: "Dev Cell. 
            2019 Jun 6. pii: S1534-5807(19)30388-0. doi: 
            10.1016/j.devcel.2019.05.016."
COMMENT     Sequence Label: pGL3-Basic-Rab18-sfGFP(N) HDR template
FEATURES             Location/Qualifiers
     source          1..5277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(756..758,759..761,762..1472)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MNSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(801..822)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1409..1430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2609..2626)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2780..3368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2860..2879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3539..4330,4331..4399))
                     /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     4162..4181
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4400..4504)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4531..4986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4618..4637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4828..4849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5117..5165
                     /note="synthetic polyadenylation signal"
     misc_feature    5179..5270
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5219..5238
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt agttgcaccg ccagaagtaa tgcacctgac ctacctgaag
       61 tctacgcctc cctttcaaga tgaattaggg tgcagagggt gctctccact ataacagagg
      121 ggaagccagc aagataatac ctgatcacac cttttataga gcactttggt taacaacatg
      181 tcttcacaaa ttaactcact taaccttcat aacaataaga agcagatgtt ctaagaagtg
      241 ttactgtgtc tctcagagat ggagaaaggc taagaaagct gggtagtatg acaaggcagt
      301 tataaggtag agctggacct aaatttaaga taatctgctt caagcacagt gcttttccca
      361 ccgcagtgca gctgttatga gcagctgccg aaataaaaca gcttgatatt ctcctgactg
      421 agtcctccca ctgactcaaa aggagggtta ggttgaggtt ggggcggagc tagacctcaa
      481 gaagaagcca acaaagtcac acacagcgct ccccacctgc gcttgcgcag gggaggaaca
      541 cgcagtccgg aaaaacgccg gggattctgg gtcgcgcagc cgtggagcta cctcaaggcg
      601 gaactgggga gtccagggct tcggcagcct gaacacccgg tgtgaaagga aggtggcaga
      661 cgtggagcgg cgcgcatgcg cagcagctca ctctgctgaa gggctgagag gcgcacccgg
      721 gcggccagct gggctcggag cggaacgggg tcaggatgaa cagcaagggc gaggagctgt
      781 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      841 gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
      901 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      961 tgcagtgctt cagccgctac cccgaccaca tgaagcgcca cgacttcttc aagtccgcca
     1021 tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc acctacaaga
     1081 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1141 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc
     1201 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1261 gccacaacgt ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1321 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgtgctga
     1381 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1441 ggatcactca cggcatggac gagctgtaca agtccggagg cggagggagc gggggaggcg
     1501 gatctggcgg aggcggatcc gacgaggacg tgctaaccac cctgaagatc ctcatcatcg
     1561 gcgagagtgg ggtgggcaag tccaggtgag gcggaggtgc gggtcgtgac gagggtggct
     1621 gggctctttc tgccccggtg gctgctgtct cctgggccgc gacgggaact gtaaactgca
     1681 gcggcgggct cgggccggct ccgctcgcgc cctcctcggc ctctgggccg cgctctcccg
     1741 cgacgtttgc tttcctctcc ccacccgcgc ctgcctggtt cccggccttt gccaggccca
     1801 agttctctgg gtcgctctcc ctcctgtagc gccgagaaaa caccattccg agggccgccg
     1861 ctcggccggc aggtttgctg cgcggagctc agggcgtggc gggctggttt gggcccgaat
     1921 ccggccgggg gcgccttgcc tcgaacctct cggggttcag gctgcctgcc accccgagac
     1981 ccgggagaca tagctttccc acttgagggt ttctgtgggg ccttctctaa atacataatg
     2041 tcctcgggaa gtttcgtgac gccttaggtc ccattcctgg cctgtttggg cgcagcttaa
     2101 tgtcgattct tttacttttt tattagtcgg ctgtgaaaga actggatctt gtgactgctg
     2161 gaaaatttcg ctttagtgta aggtccgagt gcttcagccg tgggccacct ttgaaaataa
     2221 aatgtaattg ttatttctca ttttgccaca tcctgtggag acagaagatt agtgttgtgc
     2281 tttcattttt tactttttcc ttggggaaaa taatattgac agtggagttg agtataaggt
     2341 aagacatgga tctaggtgga tgttgctaga gggctgtgat ttcatccagg tcttggaaga
     2401 gagttctcca gctgatgtca ttcttgccat ctgattatga gtaggttatg ttcaaggccg
     2461 tcctgatcgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     2521 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     2581 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2641 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2701 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2761 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2821 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2881 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2941 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3001 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3061 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3121 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3181 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3241 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3301 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3361 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3421 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3481 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3541 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3601 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3661 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3721 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3781 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3841 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3901 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3961 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4021 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4081 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4141 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4201 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4261 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4321 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4381 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4441 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4501 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     4561 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4621 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4681 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4741 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4801 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4861 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4921 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4981 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5041 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     5101 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     5161 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     5221 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgata
//
