LOCUS       sequence-417506-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty Vector for molecular cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  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 5457)
  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: PcDNA3.1-Flag
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        220..599
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        600..803
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     754..774
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     848..867
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        848..866
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      932..941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             941..964
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(995..1013)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(996..1013)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1033..1050)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1039..1263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1309..1737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1396..1415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1606..1627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1746..1766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1751..2080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1931..2066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1993..2012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2147..2941
                     /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(2201..2220)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2811..2830
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3115..3236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3152..3171)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3206..3225
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3285..3301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3285..3301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3298..3320)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3309..3325
                     /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(3333..3339,3340..3357,3358..3363))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3378..3399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3516..3533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3687..4275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3767..3786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4446..5237,5238..5306))
                     /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     5069..5088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5307..5411)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
       61 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
      121 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      181 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      241 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      301 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      361 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      421 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      481 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      541 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      601 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      661 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      721 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      781 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      841 tcgaaattaa tacgactcac tatagggaga cccaagcttg gtaccgagct cggatccact
      901 agtaacggcc gccagtgtgc tggaattcgc cgccaccatg gattataaag atgatgatga
      961 taaatgactc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     1021 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1081 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1141 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1201 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     1261 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     1321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1381 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1441 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1501 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1621 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1681 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     1741 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     1801 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1861 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     1921 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     1981 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2041 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2101 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2161 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2221 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2281 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2341 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2401 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2461 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2521 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2581 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2641 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     2701 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2761 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2821 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     2881 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     2941 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3001 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3061 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3121 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3181 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3241 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3301 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3361 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3421 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3481 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3541 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3601 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5041 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5401 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagat
//
