LOCUS       sequence-417465-        8069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ATG9A fused with 2Strep in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  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 8069)
  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: ATG9A-2STrep
FEATURES             Location/Qualifiers
     source          1..8069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3438..3447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3442..3451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3459..3482
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3519..3542
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3567..3590
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(3621..3639)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3622..3639)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3659..3676)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3665..3889
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3935..4363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4022..4041)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4232..4253
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4372..4392)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4377..4706
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4557..4692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4619..4638
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4773..5567
                     /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(4827..4846)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5437..5456
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5741..5862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5778..5797)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5832..5851
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5911..5927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5911..5927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5924..5946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5935..5951
                     /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(5959..5965,5966..5983,5984..5989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6004..6025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6142..6159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6313..6901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6393..6412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7072..7863,7864..7932))
                     /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     7695..7714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7933..8037)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag cttggtaccg
      901 agctcggatc catggcgcag tttgacactg aataccagcg cctagaggcc tcctatagtg
      961 attcaccccc aggggaggag gacctgttgg tgcacgtcgc cgaggggagc aagtcacctt
     1021 ggcaccatat tgaaaacctt gacctcttct tctctcgagt ttataatctg caccagaaga
     1081 atggcttcac atgtatgctc atcggggaga tctttgagct catgcagttc ctctttgtgg
     1141 ttgccttcac taccttcctg gtcagctgcg tggactatga catcctattt gccaacaaga
     1201 tggtgaacca cagtcttcac cctactgaac ccgtcaaggt cactctgcca gacgcctttt
     1261 tgcctgctca agtctgtagt gccaggattc aggaaaatgg ctcccttatc accatcctgg
     1321 tcattgctgg tgtcttctgg atccaccggc ttatcaagtt catctataac atttgctgct
     1381 actgggagat ccactccttc tacctgcacg ctctgcgcat ccctatgtct gcccttccgt
     1441 attgcacgtg gcaagaagtg caggcccgga tcgtgcagac gcagaaggag caccagatct
     1501 gcatccacaa acgtgagctg acagaactgg acatctacca ccgcatcctc cgtttccaga
     1561 actacatggt ggcactggtt aacaaatccc tcctgcctct gcgcttccgc ctgcctggcc
     1621 tcggggaagc tgtcttcttc acccgtggtc tcaagtacaa ctttgagctg atcctcttct
     1681 ggggacctgg ctctctgttt ctcaatgaat ggagcctcaa ggccgagtac aaacgtgggg
     1741 ggcaacggct agagctggcc cagcgcctca gcaaccgcat cctgtggatt ggcatcgcta
     1801 acttcctgct gtgccccctc atcctcatat ggcaaatcct ctatgccttc ttcagctatg
     1861 ctgaggtgct gaagcgggag ccgggggccc tgggagcacg ctgctggtca ctctatggcc
     1921 gctgctacct ccgccacttc aacgagctgg agcacgagct gcagtcccgc ctcaaccgtg
     1981 gctacaagcc cgcctccaag tacatgaatt gcttcttgtc acctcttttg acactgctgg
     2041 ccaagaatgg agccttcttc gctggctcca tcctggctgt gcttattgcc ctcaccattt
     2101 atgacgaaga tgtgttggct gtggaacatg tgctgaccac cgtcacactc ctgggggtca
     2161 ccgtgaccgt gtgcaggtcc tttatcccgg accagcacat ggtgttctgc cctgagcagc
     2221 tgctccgcgt gatcctcgct cacatccact acatgcctga ccactggcag ggtaatgccc
     2281 accgctcgca gacccgggac gagtttgccc agctcttcca gtacaaggca gtgttcattt
     2341 tggaagagtt gctgagcccc attgtcacac ccctcatcct catcttctgc ctgcgcccac
     2401 gggccctgga gattatagac ttcttccgaa acttcaccgt ggaggtcgtt ggtgtgggag
     2461 atacctgctc ctttgctcag atggatgttc gccagcatgg tcatccccag tggctatctg
     2521 ctgggcagac agaggcctca gtgtaccagc aagctgagga tggaaagaca gagttgtcac
     2581 tcatgcactt tgccatcacc aaccctggct ggcagccacc acgtgagagc acagccttcc
     2641 taggcttcct caaggagcag gttcagcggg atggagcagc tgctagcctc gcccaagggg
     2701 gtctgctccc tgaaaatgcc ctctttacgt ctatccagtc cttacaatct gagtctgagc
     2761 ccctgagcct tatcgcaaat gtggtagctg gctcatcctg ccggggccct ccactgccca
     2821 gagacctgca gggctccagg cacagggctg aagtcgcctc tgccctgcgc tccttctccc
     2881 cgctgcaacc cgggcaggcg cccacaggcc gggctcacag caccatgaca ggctctgggg
     2941 tggatgccag gacagccagc tccgggagca gcgtgtggga aggacagctg cagagcctgg
     3001 tgctgtcaga atatgcatcc acagagatga gcctgcatgc cctctatatg caccagctcc
     3061 acaagcagca ggcccaggct gaacctgagc ggcatgtatg gcaccgccgg gagagtgatg
     3121 agagtggaga aagcgcccct gatgaagggg gagagggcgc ccgggccccc cagtctatcc
     3181 ctcgctctgc tagctatccc tgtgcagcac cccggcctgg agctcctgag accaccgccc
     3241 tgcatggggg cttccagagg cgctacggtg gcatcacaga tcctggcaca gtgcccaggg
     3301 ttccctctca tttctctcgg ctgcctcttg gagggtgggc agaagatggg cagtcggcat
     3361 caaggcaccc tgagcccgtg cccgaagagg gctcggagga tgagctaccc cctcaggtgc
     3421 acaaggtaga attcgccgcc accatggcgg ccgccagctg gagccaccct cagttcgaga
     3481 agggaggagg aagcggcgga ggcagcggag gaggaagctg gagccacccg cagttcgaga
     3541 aatgactcga gcatgcatct agaggctgga gccacccgca gttcgagaaa tgactcgagc
     3601 atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     3661 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3721 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3781 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3841 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3901 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3961 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4021 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4081 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4141 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4201 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4261 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4321 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4381 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4441 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4501 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4561 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4621 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4681 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4741 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4801 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4861 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4921 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4981 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5041 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5101 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5161 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5221 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5281 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5341 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5401 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5461 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5521 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5581 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5641 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5701 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5761 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5821 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5881 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5941 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6001 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6061 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6121 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6181 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6241 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6301 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6361 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6421 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6481 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6541 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6601 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6661 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6721 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6781 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6841 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6901 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6961 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7021 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7141 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7201 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7261 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7501 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7561 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7681 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7741 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7921 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8041 atttccccga aaagtgccac ctgacgtcg
//
