LOCUS       sequence-418124-        7900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ATP6V0A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7900)
  AUTHORS   Li J, Gui Q, Liang FX, Sall J, Zhang Q, Duan Y, Dhabaria A, Askenazi
            M, Ueberheide B, Stapleford KA, Pagano M
  TITLE     The REEP5/TRAM1 complex binds SARS-CoV-2 NSP3 and promotes virus 
            replication.
  JOURNAL   J Virol. 2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23.
  PUBMED    37768083
REFERENCE   2  (bases 1 to 7900)
  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: "J Virol. 
            2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23."
COMMENT     Sequence Label: Flag-V0A
FEATURES             Location/Qualifiers
     source          1..7900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..946
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3492..3509)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3498..3722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3768..4196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3855..3874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4065..4086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4205..4225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4210..4539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4390..4525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4452..4471
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4606..5400
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4660..4679)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5270..5289
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5574..5695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5611..5630)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5665..5684
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5744..5760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5744..5760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5757..5779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5768..5784
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5792..5798,5799..5816,5817..5822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5837..5858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5975..5992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6146..6734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6226..6245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6904..7695,7696..7764))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7527..7546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7765..7869)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttacca tggactacaa ggacgacgat gacaaggtac caggatctat
      961 gggggagctt ttccggagtg aagaaatgac actggcccag ctttttctac agtcagaggc
     1021 tgcttattgt tgtgtcagtg aattaggaga acttggaaag gttcagtttc gtgacttaaa
     1081 tccagatgtg aatgttttcc aacggaaatt tgtgaatgaa gttagaagat gtgaagaaat
     1141 ggatcgaaag cttcgatttg ttgagaaaga gataagaaaa gctaacattc cgattatgga
     1201 caccggtgaa aacccagagg ttcccttccc ccgggacatg attgacttag aggccaattt
     1261 tgagaagatt gaaaatgaac tgaaggaaat caacacaaac caggaagctc tgaagagaaa
     1321 cttcctggaa ctgaccgaat taaaatttat acttcgcaaa actcagcaat tttttgatga
     1381 gatggcggat ccagacttgt tggaagagtc ctcatccctc ttggagccaa gtgagatggg
     1441 aagaggcact cctttaagac ttggcttcgt ggctggtgtc attaaccggg agcgcatccc
     1501 tacttttgag cgcatgcttt ggcgggtatg ccggggaaat gtgttcctgc gacaggctga
     1561 aatcgagaac cccctggagg atcctgtgac tggcgactac gtgcacaagt ctgtgtttat
     1621 cattttcttc caaggtgatc agctgaaaaa cagagtcaag aaaatctgtg aagggttccg
     1681 agcctcactc tatccctgtc ctgagacacc acaggagagg aaggaaatgg cttctggagt
     1741 gaataccagg attgatgatc tccaaatggt tctgaatcaa acggaggatc accgccagag
     1801 ggttctgcag gcagctgcta agaacatccg tgtctggttc atcaaagtgc ggaagatgaa
     1861 ggccatctat cacaccctga acctgtgcaa catagatgtg actcagaaat gcttgattgc
     1921 agaggtctgg tgccctgtca ccgaccttga ctccatccag tttgcactca gaaggggcac
     1981 ggaacacagt ggttccactg taccttccat tttgaacagg atgcagacaa accagactcc
     2041 cccaacctat aacaaaacca acaagtttac ctatggcttt cagaacatag tagatgctta
     2101 tggaattgga acttaccgag agataaatcc agctccgtat actattatca cgttcccttt
     2161 tctatttgct gtgatgtttg gagacttcgg tcatggcatt ttaatgaccc tttttgctgt
     2221 gtggatggta ctgagggaga gccggatcct ttcccagaag aatgagaatg agatgtttag
     2281 cactgtgttc agtggtcgat acattatttt attgatgggt gtgttctcca tgtacactgg
     2341 cctcatctac aatgattgct tttccaagtc tcttaatatc tttgggtcat cctggagtgt
     2401 acggccgatg tttacttata attggactga agagacgctt cgggggaacc ctgttctaca
     2461 gctgaaccca gccctccctg gagtgtttgg tggaccatac ccttttggca ttgatccaat
     2521 ttggaacatt gctaccaata aactgacgtt cttgaactcc tttaagatga agatgtctgt
     2581 tatccttggt atcatccata tgctgtttgg agtcagcctg agtctgttca accatatcta
     2641 tttcaagaag cccctgaata tctactttgg atttattcct gaaataatct tcatgacctc
     2701 tttgtttggc tatttggtta tccttatttt ttacaagtgg acggcctatg atgctcatac
     2761 ctctgagaat gcaccaagcc ttctgatcca tttcataaac atgttcctct tttcctaccc
     2821 agagtctggt tattcaatgt tgtattctgg acagaaagga attcagtgtt tcctggtagt
     2881 ggttgcacta ctgtgtgtac cttggatgct gctgtttaaa ccattggtcc ttcgccgtca
     2941 gtatttgagg agaaagcatt tgggaactct caactttggt gggatcaggg tgggcaacgg
     3001 accgacagag gaggatgctg agattattca gcatgaccag ctctccaccc actcagagga
     3061 cgcagacgag tttgactttg gggacaccat ggtccaccag gccatccaca ccatcgagta
     3121 ctgcctgggc tgcatctcca acactgcctc ctacttgcgg ctctgggccc tcagcctcgc
     3181 tcatgcgcag ctgtctgagg tgctttggac catggtgatc cacatcggcc tgagcgtgaa
     3241 gagcttggcg ggaggtttgg tgctgttctt cttcttcact gcctttgcca ccctgaccgt
     3301 ggccatcctc ctgatcatgg agggcctctc ggcctttctc cacgcactgc gcttacactg
     3361 ggttgagttc cagaataaat tctacagcgg gaccggtttc aagttcttac ccttctcctt
     3421 cgagcatatt cgggaaggga agtttgaaga gtgactcgag tctagagggc ccgtttaaac
     3481 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3541 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3601 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3661 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3721 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3781 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3841 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3901 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3961 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4021 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4081 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4141 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4441 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4561 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4621 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4681 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4741 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4801 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4861 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4921 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4981 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5041 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5101 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5161 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5221 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5281 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5341 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5401 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5461 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5521 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5581 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5641 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5701 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5761 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5821 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5881 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5941 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6721 aaaaaggatc tcaagaagat cctttgatct tttctacggg tctgacgctc agtggaacga
     6781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
