LOCUS       sequence-501956-        4649 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Transient mammalian expression of SAR1BH79G-3xFLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4649)
  AUTHORS   Fuller DM, Wu Y, Schueder F, Rasool B, Nag S, Korfhage JL, 
            Garcia-Milian R, Melnyk KD, Bewersdorf J, De Camilli P, Melia TJ
  TITLE     ATG2A engages RAB1A and ARFGAP1 positive membranes during 
            autophagosome biogenesis
  JOURNAL   eLife 2025 14:RP107316
REFERENCE   2  (bases 1 to 4649)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife 2025 
            14:RP107316"
COMMENT     Sequence Label: SAR1BH79G-3xFLAG
FEATURES             Location/Qualifiers
     source          1..4649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             604..669
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    794..915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(831..850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     885..904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(922..1377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1059..1080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1271..1290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1404..1508
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1510..1867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1510..1701
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1533..1553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1718..1853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1902..2696
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1902..2696
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1956..1975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2566..2585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2884..2903)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2928..2975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3304..3892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3793..3812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4067..4370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4371..4574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4525..4545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atgtccttca tatttgactg gatttacagt ggtttcagca gtgtgctgca gtttctagga
       61 ctgtataaaa aaactggtaa actggtattc cttggattgg acaatgctgg gaaaaccacg
      121 ttgctacaca tgctgaaaga tgacagactt ggccagcacg tcccaacact acatcccact
      181 tcagaagaac tcactattgc tggcatgact tttacaactt ttgatctggg tgggggcatc
      241 caagcccgaa gagtatggaa aaattacctt cctgctatca atggcattgt atttcttgtg
      301 gattgtgcag accatgaaag gctgttagaa tcaaaagaag aacttgattc actaatgaca
      361 gatgaaacca ttgccaatgt gcctatatta attcttggaa ataagattga cagacccgaa
      421 gccatcagtg aagagaggtt acgagagatg tttggcttat atgggcagac aacaggaaag
      481 ggcagtgtgt cactgaagga gctgaatgcc agacctctgg aagtgttcat gtgcagtgtg
      541 ctgaaaaggc aaggctatgg agaaggcttc cgctggatgg cacagtacat cgatggggat
      601 ccagactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
      661 gatgacaagt aggcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
      721 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
      781 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      841 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      901 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
      961 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1021 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1081 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1141 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1201 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1261 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1321 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     1381 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1441 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1501 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     1561 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1621 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1681 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     1741 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     1801 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     1861 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     1921 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     1981 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2041 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     2101 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2161 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2221 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2281 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     2341 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     2401 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     2461 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     2521 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     2581 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     2641 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     2701 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     2761 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     2821 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     2881 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     2941 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3001 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3061 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3121 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     3181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3961 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     4021 aatcaattac ggggtcatta gttaatagcc catatatgga gttccgcgtt acataactta
     4081 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4141 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4201 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4261 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4321 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4381 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4441 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4501 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     4561 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
     4621 cgagctcaag cttcgaattc accgccacc
//
