LOCUS       sequence-207759-        6570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inactive 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-SAC1C389S-HLx8
FEATURES             Location/Qualifiers
     source          1..6570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        70..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        374..577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     528..548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      616..625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             622..1329
                     /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(767..785)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1035..1054
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3367..3488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3404..3423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3458..3477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3495..3950)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3632..3653)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3844..3863
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3977..4081
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4083..4440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4083..4274
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4106..4126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4291..4426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4475..5269
                     /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             4475..5269
                     /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(4529..4548)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5139..5158
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5457..5476)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5501..5548
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5877..6465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6366..6385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
       61 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      121 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      181 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      241 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      301 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      361 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      421 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      481 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      541 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      601 ctagcgctac cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc
      661 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      721 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      781 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
      841 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
      901 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      961 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1021 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1081 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     1141 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1201 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     1261 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcaat ggcgacggcg
     1381 gcctacgagc agctgaagct gcatatcaca cctgaaaaat tttatgtgga agcttgtgat
     1441 gatggagcag atgacgtact taccattgac cgtgtgtcca cagaggttac ccttgcagtc
     1501 aagaaagatg ttcctccttc agctgtcaca agaccaatat ttggtatact gggcacaatc
     1561 catctggtgg caggtaatta tcttatagtc attaccaaaa agataaaagt aggtgaattt
     1621 ttcagtcatg tagtctggaa agcaacagat tttgatgtcc tttcttataa gaagacaatg
     1681 ttgcacttaa ctgatattca gttacaagat aataaaacct tcctagcgat gctaaaccat
     1741 gtcttgaatg tggatggatt ttacttttca acaacatatg atttgaccca tactttgcag
     1801 cggctatcca acactagtcc tgaattccaa gaaatgagtc tcttggaaag ggcagatcag
     1861 cggtttgtat ggaatggtca tcttctaaga gaactttctg cacagccaga ggttcatcgg
     1921 tttgcccttc cagtgttaca tggctttatt accatgcatt catgttctat taatggaaaa
     1981 tactttgatt ggattctcat ctcgaggagg agctgtttca gagctggtgt gcgctattat
     2041 gtaagaggaa ttgattcgga aggccatgca gctaactttg tagaaacaga acaaattgtg
     2101 cactacaatg ggagcaaagc ttcgtttgta cagactcgag gatcaatacc tgttttctgg
     2161 tcccaaagac caaacctcaa gtacaaacca ctgccacaga tcagcaaagt agcaaatcac
     2221 atggacggtt tccaaaggca ttttgattcc caagtaatta tttatggaaa acaagttata
     2281 atcaatctga ttaaccagaa gggctcggag aagccacttg agcagacatt tgcaacaatg
     2341 gtgtcttcct tgggaagtgg aatgatgaga tacattgcct ttgacttcca taaggaatgt
     2401 aaaaatatga gatgggatcg actaagtatt ttattggatc aggtagcaga aatgcaagat
     2461 gaattaagtt attttctagt ggactctgct ggccaggtgg tggcaaacca ggaaggcgtg
     2521 ttccgaagca atagcatgga ttgtctagat agaaccaatg tgatccagag tttgttagct
     2581 cgtcgttcac ttcaggccca acttcagaga ctaggagttt tgcatgtggg acaaaagctt
     2641 gaagaacaag atgaatttga gaagattttc aaaaatgcct gggctgacaa cgcaaatgct
     2701 tgtgccaagc aatatgcggg aactggtgcc ttgaagactg actttaccag aactggaaag
     2761 agaactcatt tgggacttat aatggatggc tggaactcaa tgatacgata ttataagaac
     2821 aacttttccg atggatttag acaagattcc atagacttat ttcttggaaa ctattcagtg
     2881 gatgaattag aatctcatag tcctttaagt gttccaaggg actggaaaga agccgctgct
     2941 agagaagctg ccgcaagaga ggccgcagcc agagaggctg ctgccaggga agcagcagct
     3001 cgggaagccg cagcacgcga ggctgcagca agagaagcag ccgcacgctt cctggctttg
     3061 cctattatca tggttgttgc cttttcaatg tgcattatct gtttgcttat ggctggtgac
     3121 acttggacag aaacactggc ctatgtgctc ttctggggag ttgcaagcat tggaacattt
     3181 tttatcattc tttacaatgg caaagatttt gtcgatgctc ccagactggt ccagaaagaa
     3241 aagatagact gaagataact gatcataatc agccatacca catttgtaga ggttttactt
     3301 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3361 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3421 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3481 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3541 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3601 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3661 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3721 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3781 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3841 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3901 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3961 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4021 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4081 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4141 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4201 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4261 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4321 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4381 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4441 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4501 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4561 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4621 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4681 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4741 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4801 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4861 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4921 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4981 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5041 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5101 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5161 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5221 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5281 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5341 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5401 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5461 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5521 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5581 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5641 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5701 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5761 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5821 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5881 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5941 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6001 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6061 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6121 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6181 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6241 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6301 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6361 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6421 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6481 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6541 tcccctgatt ctgtggataa ccgtattacc
//
