LOCUS       sequence-254784-        4482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HDR tempalte for tagging of endogenous human ACSL3 C-terminus with 
            sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4482)
  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 4482)
  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-ACSL3-sfGFP(C) HDR template
FEATURES             Location/Qualifiers
     source          1..4482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(615..617,618..620,621..1334)
                     /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="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(660..681)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1268..1289
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1814..1831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1985..2573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2065..2084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2744..3535,3536..3604))
                     /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     3367..3386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3605..3709)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3736..4191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3823..3842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4033..4054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4322..4370
                     /note="synthetic polyadenylation signal"
     misc_feature    4384..4475
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4424..4443
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgtta cttattttag agacaaggtc tcgctatgtt accgaggctg gccttgaatt
       61 cctggcctca agcgatcctc ctgcctgagc tttcagagtt actgggatta caggcatgag
      121 ccatcatgcc tggctctaac tcgttttcaa tgacagaata tgtattctat tttgttccta
      181 tatcatattt tatttaactc attttcctct tgatggggac ttgttaaata gtttattgtt
      241 ttacttatgt ttgtattttg aggaatagat tctttagaag tgacattcat ggttcacttg
      301 ttggttgtgc attttgatgg atacgcaaaa agtggggaag ttacaccatt tgctaagaac
      361 ttaaatacct tttcttcttt tttttttttt taatcatctt agcaagtctg gaaaagtttg
      421 aaattccagt aaaaattcgt ttgagtcctg aaccgtggac ccctgaaact ggtctggtga
      481 cagatgcctt caagctgaaa cgcaaagagc ttaaaacaca ttaccaggcg gacattgagc
      541 gaatgtatgg aagaaaagga tctggcggag gagggagcgg gggaggcgga tctggcggag
      601 gcggatccct cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      661 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg
      721 gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      781 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      841 ccgaccacat gaagcgccac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      901 agcgcaccat cagcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg
      961 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1021 acatcctggg gcacaagctg gagtacaact tcaacagcca caacgtctat atcaccgccg
     1081 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg gaggacggca
     1141 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1201 tgcccgacaa ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc
     1261 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcatggacg
     1321 agctgtacaa gtaattattc tcttctggca tcagtttgct acagtgagct cagatcaaat
     1381 aggaaaatac ttgaaatgca tgtctcaagc tgcaaggcaa actccattcc tcatattaaa
     1441 ctattacttc tcatgacgtc accattttta actgacagga ttagtaaaac attaagacag
     1501 caaacttgtg tctgtctctt ctttcatttt ccccgccacc aacttacttt accacctatg
     1561 actgtacttg tcagtatgag aatttttctg aatcatattg gggaagcagt gattttaaaa
     1621 cctcaagttt ttaaacatga tttatatgtt ctgtataatg ttcagtttgt aacgtcgacc
     1681 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     1741 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     1801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2401 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3721 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3781 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3841 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3901 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3961 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4021 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4081 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4141 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     4201 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4261 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     4321 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     4381 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     4441 ccccagtgca agtgcaggtg ccagaacatt tctctatcga ta
//
