LOCUS       sequence-501955-        5291 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Transient mammalian expression of SAR1B-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5291)
  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 5291)
  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: SAR1B-BFP
FEATURES             Location/Qualifiers
     source          1..5291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             616..1314
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1436..1557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1473..1492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1527..1546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1564..2019)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1701..1722)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1913..1932
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2046..2150
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2152..2509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2152..2343
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2175..2195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2360..2495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2544..3338
                     /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             2544..3338
                     /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(2598..2617)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3208..3227
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3526..3545)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3570..3617
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3946..4534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4435..4454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4709..5012
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5013..5216
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5167..5187
                     /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 tgggcacatc
      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 ccaccggtcg ccaccatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
      661 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
      721 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
      781 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
      841 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
      901 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      961 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     1021 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     1081 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag
     1141 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     1201 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     1261 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taatgcggcc
     1321 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1381 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     1561 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1621 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1681 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1741 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1801 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1861 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1921 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1981 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2041 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2101 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2161 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2221 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2281 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2341 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2401 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2461 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     2521 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2581 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2641 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2701 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     2761 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2821 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2881 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2941 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3001 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3061 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3121 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3181 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3241 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3301 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3361 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3421 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3481 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     3541 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3601 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3661 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3721 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3781 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3841 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3901 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3961 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4021 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4081 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4141 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4201 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4261 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4321 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4381 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4441 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4501 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4561 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4621 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     4681 tagttaatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4741 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4801 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4861 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4921 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4981 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     5041 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     5101 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     5161 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     5221 tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat
     5281 tcaccgccac c
//
