LOCUS       sequence-417466-        7211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ATG9A fused with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7211)
  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 7211)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..7211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        351..554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     505..525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3137..3146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3143..3145,3146..3148,3149..3862)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3188..3209)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3449..3468)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3796..3817
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3985..4106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4022..4041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4076..4095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4113..4568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4250..4271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4462..4481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4595..4699
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4701..5058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4701..4892
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4724..4744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4909..5044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5093..5887
                     /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             5093..5887
                     /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(5147..5166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5757..5776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6075..6094)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6119..6166
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6495..7083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6984..7003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
       61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      541 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      601 cgagatggcg cagtttgaca ctgaatacca gcgcctagag gcctcctata gtgattcacc
      661 cccaggggag gaggacctgt tggtgcacgt cgccgagggg agcaagtcac cttggcacca
      721 tattgaaaac cttgacctct tcttctctcg agtttataat ctgcaccaga agaatggctt
      781 cacatgtatg ctcatcgggg agatctttga gctcatgcag ttcctctttg tggttgcctt
      841 cactaccttc ctggtcagct gcgtggacta tgacatccta tttgccaaca agatggtgaa
      901 ccacagtctt caccctactg aacccgtcaa ggtcactctg ccagacgcct ttttgcctgc
      961 tcaagtctgt agtgccagga ttcaggaaaa tggctccctt atcaccatcc tggtcattgc
     1021 tggtgtcttc tggatccacc ggcttatcaa gttcatctat aacatttgct gctactggga
     1081 gatccactcc ttctacctgc acgctctgcg catccctatg tctgcccttc cgtattgcac
     1141 gtggcaagaa gtgcaggccc ggatcgtgca gacgcagaag gagcaccaga tctgcatcca
     1201 caaacgtgag ctgacagaac tggacatcta ccaccgcatc ctccgtttcc agaactacat
     1261 ggtggcactg gttaacaaat ccctcctgcc tctgcgcttc cgcctgcctg gcctcgggga
     1321 agctgtcttc ttcacccgtg gtctcaagta caactttgag ctgatcctct tctggggacc
     1381 tggctctctg tttctcaatg aatggagcct caaggccgag tacaaacgtg gggggcaacg
     1441 gctagagctg gcccagcgcc tcagcaaccg catcctgtgg attggcatcg ctaacttcct
     1501 gctgtgcccc ctcatcctca tatggcaaat cctctatgcc ttcttcagct atgctgaggt
     1561 gctgaagcgg gagccggggg ccctgggagc acgctgctgg tcactctatg gccgctgcta
     1621 cctccgccac ttcaacgagc tggagcacga gctgcagtcc cgcctcaacc gtggctacaa
     1681 gcccgcctcc aagtacatga attgcttctt gtcacctctt ttgacactgc tggccaagaa
     1741 tggagccttc ttcgctggct ccatcctggc tgtgcttatt gccctcacca tttatgacga
     1801 agatgtgttg gctgtggaac atgtgctgac caccgtcaca ctcctggggg tcaccgtgac
     1861 cgtgtgcagg tcctttatcc cggaccagca catggtgttc tgccctgagc agctgctccg
     1921 cgtgatcctc gctcacatcc actacatgcc tgaccactgg cagggtaatg cccaccgctc
     1981 gcagacccgg gacgagtttg cccagctctt ccagtacaag gcagtgttca ttttggaaga
     2041 gttgctgagc cccattgtca cacccctcat cctcatcttc tgcctgcgcc cacgggccct
     2101 ggagattata gacttcttcc gaaacttcac cgtggaggtc gttggtgtgg gagatacctg
     2161 ctcctttgct cagatggatg ttcgccagca tggtcatccc cagtggctat ctgctgggca
     2221 gacagaggcc tcagtgtacc agcaagctga ggatggaaag acagagttgt cactcatgca
     2281 ctttgccatc accaaccctg gctggcagcc accacgtgag agcacagcct tcctaggctt
     2341 cctcaaggag caggttcagc gggatggagc agctgctagc ctcgcccaag ggggtctgct
     2401 ccctgaaaat gccctcttta cgtctatcca gtccttacaa tctgagtctg agcccctgag
     2461 ccttatcgca aatgtggtag ctggctcatc ctgccggggc cctccactgc ccagagacct
     2521 gcagggctcc aggcacaggg ctgaagtcgc ctctgccctg cgctccttct ccccgctgca
     2581 acccgggcag gcgcccacag gccgggctca cagcaccatg acaggctctg gggtggatgc
     2641 caggacagcc agctccggga gcagcgtgtg ggaaggacag ctgcagagcc tggtgctgtc
     2701 agaatatgca tccacagaga tgagcctgca tgccctctat atgcaccagc tccacaagca
     2761 gcaggcccag gctgaacctg agcggcatgt atggcaccgc cgggagagtg atgagagtgg
     2821 agaaagcgcc cctgatgaag ggggagaggg cgcccgggcc ccccagtcta tccctcgctc
     2881 tgctagctat ccctgtgcag caccccggcc tggagctcct gagaccaccg ccctgcatgg
     2941 gggcttccag aggcgctacg gtggcatcac agatcctggc acagtgccca gggttccctc
     3001 tcatttctct cggctgcctc ttggagggtg ggcagaagat gggcagtcgg catcaaggca
     3061 ccctgagccc gtgcccgaag agggctcgga ggatgagcta ccccctcagg tgcacaaggt
     3121 agcggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     3181 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3241 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3301 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3361 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3421 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3481 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3541 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3601 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3661 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3721 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     3781 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3841 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     3901 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3961 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4081 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     4141 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4201 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4261 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4321 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4381 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4441 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4501 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4561 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4681 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4741 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4801 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4861 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4921 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4981 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5041 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5101 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5161 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5221 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5281 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5341 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5401 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5461 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5521 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5581 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5641 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5701 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5761 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5821 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5881 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5941 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6001 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6061 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6121 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6181 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6241 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6301 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6361 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6421 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6481 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6541 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6601 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6661 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6721 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6781 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6841 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6901 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6961 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7021 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7081 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7141 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     7201 ttattaatag t
//
