LOCUS       sequence-203958-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3358..3479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3395..3414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3449..3468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3486..3941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3623..3644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3835..3854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3968..4072
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4074..4431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4074..4265
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4097..4117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4282..4417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4466..5260
                     /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             4466..5260
                     /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(4520..4539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5130..5149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5448..5467)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5492..5539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5868..6456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6357..6376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tcaagcttcg aattctgcaa tggcgacggc ggcctacgag
     1381 cagctgaagc tgcatatcac acctgaaaaa ttttatgtgg aagcttgtga tgatggagca
     1441 gatgacgtac ttaccattga ccgtgtgtcc acagaggtta cccttgcagt caagaaagat
     1501 gttcctcctt cagctgtcac aagaccaata tttggtatac tgggcacaat ccatctggtg
     1561 gcaggtaatt atcttatagt cattaccaaa aagataaaag taggtgaatt tttcagtcat
     1621 gtagtctgga aagcaacaga ttttgatgtc ctttcttata agaagacaat gttgcactta
     1681 actgatattc agttacaaga taataaaacc ttcctagcga tgctaaacca tgtcttgaat
     1741 gtggatggat tttacttttc aacaacatat gatttgaccc atactttgca gcggctatcc
     1801 aacactagtc ctgaattcca agaaatgagt ctcttggaaa gggcagatca gcggtttgta
     1861 tggaatggtc atcttctaag agaactttct gcacagccag aggttcatcg gtttgccctt
     1921 ccagtgttac atggctttat taccatgcat tcatgttcta ttaatggaaa atactttgat
     1981 tggattctca tctcgaggag gagctgtttc agagctggtg tgcgctatta tgtaagagga
     2041 attgattcgg aaggccatgc agctaacttt gtagaaacag aacaaattgt gcactacaat
     2101 gggagcaaag cttcgtttgt acagactcga ggatcaatac ctgttttctg gtcccaaaga
     2161 ccaaacctca agtacaaacc actgccacag atcagcaaag tagcaaatca catggacggt
     2221 ttccaaaggc attttgattc ccaagtaatt atttatggaa aacaagttat aatcaatctg
     2281 attaaccaga agggctcgga gaagccactt gagcagacat ttgcaacaat ggtgtcttcc
     2341 ttgggaagtg gaatgatgag atacattgcc tttgacttcc ataaggaatg taaaaatatg
     2401 agatgggatc gactaagtat tttattggat caggtagcag aaatgcaaga tgaattaagt
     2461 tattttctag tggactctgc tggccaggtg gtggcaaacc aggaaggcgt gttccgaagc
     2521 aatagcatgg attgtctaga tagaaccaat gtgatccaga gtttgttagc tcgtcgttca
     2581 cttcaggccc aacttcagag actaggagtt ttgcatgtgg gacaaaagct tgaagaacaa
     2641 gatgaatttg agaagattta caaaaatgcc tgggctgaca acgcaaatgc ttgtgccaag
     2701 caatatgcgg gaactggtgc cttgaagact gactttacca gaactggaaa gagaactcat
     2761 ttgggactta taatggatgg ctggaactca atgatacgat attataagaa caacttttcc
     2821 gatggattta gacaagattc catagactta tttcttggaa actattcagt ggatgaatta
     2881 gaatctcata gtcctttaag tgttccaagg gactggaaag aagccgctgc tagagaagct
     2941 gccgcaagag aggccgcagc cagagaggct gctgccaggg aagcagcagc tcgggaagcc
     3001 gcagcacgcg aggctgcagc aagagaagca gccgcacgct tcctggcttt gcctattatc
     3061 atggttgttg ccttttcaat gtgcattatc tgtttgctta tggctggtga cacttggaca
     3121 gaaacactgg cctatgtgct cttctgggga gttgcaagca ttggaacatt ttttatcatt
     3181 ctttacaatg gcaaagattt tgtcgatgct cccagactgg tccagaaaga aaagatagac
     3241 tgaagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3301 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3361 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3421 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3481 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3541 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3601 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3661 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3721 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3781 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3841 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3901 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3961 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4021 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4081 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4261 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4321 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4381 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4441 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4501 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4561 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4621 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4681 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4741 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4801 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4861 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4921 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4981 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5041 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5101 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5161 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5221 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5281 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5341 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5401 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5461 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5521 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5581 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5641 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5701 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5761 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5821 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5881 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5941 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6001 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6061 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6121 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6181 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6241 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6301 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6361 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6421 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6481 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6541 tctgtggata accgtattac cgccatgcat
//
