LOCUS       sequence-207735-        6570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SAC1 with extended linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6570)
  AUTHORS   Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR
  TITLE     SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic 
            reticulum to maintain a steep chemical gradient with donor 
            membranes.
  JOURNAL   Elife. 2018 Feb 20;7. pii: 35588. doi: 10.7554/eLife.35588.
  PUBMED    29461204
REFERENCE   2  (bases 1 to 6570)
  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: "Elife."; 
            date: "2018-02-20"; volume: "7. pii: 35588. doi"; pages: " 
            10.7554/eLife.35588"
COMMENT     Sequence Label: mCherry-SAC1-HLx8
FEATURES             Location/Qualifiers
     source          1..6570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      613..622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             619..1326
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(764..782)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1032..1051
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3364..3485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3401..3420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3455..3474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3492..3947)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3629..3650)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3841..3860
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3974..4078
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4080..4437
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4080..4271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4103..4123)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4288..4423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4472..5266
                     /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             4472..5266
                     /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(4526..4545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5136..5155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5454..5473)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5498..5545
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5874..6462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6363..6382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
      661 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      721 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      781 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      841 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
      901 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
      961 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     1021 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     1081 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     1141 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     1201 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     1261 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1321 tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcaatggc gacggcggcc
     1381 tacgagcagc tgaagctgca tatcacacct gaaaaatttt atgtggaagc ttgtgatgat
     1441 ggagcagatg acgtacttac cattgaccgt gtgtccacag aggttaccct tgcagtcaag
     1501 aaagatgttc ctccttcagc tgtcacaaga ccaatatttg gtatactggg cacaatccat
     1561 ctggtggcag gtaattatct tatagtcatt accaaaaaga taaaagtagg tgaatttttc
     1621 agtcatgtag tctggaaagc aacagatttt gatgtccttt cttataagaa gacaatgttg
     1681 cacttaactg atattcagtt acaagataat aaaaccttcc tagcgatgct aaaccatgtc
     1741 ttgaatgtgg atggatttta cttttcaaca acatatgatt tgacccatac tttgcagcgg
     1801 ctatccaaca ctagtcctga attccaagaa atgagtctct tggaaagggc agatcagcgg
     1861 tttgtatgga atggtcatct tctaagagaa ctttctgcac agccagaggt tcatcggttt
     1921 gcccttccag tgttacatgg ctttattacc atgcattcat gttctattaa tggaaaatac
     1981 tttgattgga ttctcatctc gaggaggagc tgtttcagag ctggtgtgcg ctattatgta
     2041 agaggaattg attcggaagg ccatgcagct aactttgtag aaacagaaca aattgtgcac
     2101 tacaatggga gcaaagcttc gtttgtacag actcgaggat caatacctgt tttctggtcc
     2161 caaagaccaa acctcaagta caaaccactg ccacagatca gcaaagtagc aaatcacatg
     2221 gacggtttcc aaaggcattt tgattcccaa gtaattattt atggaaaaca agttataatc
     2281 aatctgatta accagaaggg ctcggagaag ccacttgagc agacatttgc aacaatggtg
     2341 tcttccttgg gaagtggaat gatgagatac attgcctttg acttccataa ggaatgtaaa
     2401 aatatgagat gggatcgact aagtatttta ttggatcagg tagcagaaat gcaagatgaa
     2461 ttaagttatt ttctagtgga ctctgctggc caggtggtgg caaaccagga aggcgtgttc
     2521 cgaagcaatt gcatggattg tctagataga accaatgtga tccagagttt gttagctcgt
     2581 cgttcacttc aggcccaact tcagagacta ggagttttgc atgtgggaca aaagcttgaa
     2641 gaacaagatg aatttgagaa gattttcaaa aatgcctggg ctgacaacgc aaatgcttgt
     2701 gccaagcaat atgcgggaac tggtgccttg aagactgact ttaccagaac tggaaagaga
     2761 actcatttgg gacttataat ggatggctgg aactcaatga tacgatatta taagaacaac
     2821 ttttccgatg gatttagaca agattccata gacttatttc ttggaaacta ttcagtggat
     2881 gaattagaat ctcatagtcc tttaagtgtt ccaagggact ggaaagaagc cgctgctaga
     2941 gaagctgccg caagagaggc cgcagccaga gaggctgctg ccagggaagc agcagctcgg
     3001 gaagccgcag cacgcgaggc tgcagcaaga gaagcagccg cacgcttcct ggctttgcct
     3061 attatcatgg ttgttgcctt ttcaatgtgc attatctgtt tgcttatggc tggtgacact
     3121 tggacagaaa cactggccta tgtgctcttc tggggagttg caagcattgg aacatttttt
     3181 atcattcttt acaatggcaa agattttgtc gatgctccca gactggtcca gaaagaaaag
     3241 atagactgaa gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3301 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3361 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3421 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3481 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3541 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3601 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3661 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3721 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3781 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3841 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3901 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4081 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4261 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4321 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4441 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4501 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4561 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4621 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4681 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4741 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4801 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4861 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4921 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4981 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5041 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5101 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5161 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5221 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5281 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5341 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5401 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5461 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5521 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5581 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5641 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5701 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5761 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5821 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5881 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5941 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6001 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6061 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6421 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6481 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6541 cctgattctg tggataaccg tattaccgcc
//
