LOCUS       sequence-467513-        6254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF 
            (enhanced Magnet, acidic, fast), V5, nuclear export sequence, MAVS 
            residues 500-540 for targeting to outer mitochondrial membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6254)
  AUTHORS   Zhang Z, Xu A, Bai Y, Chen Y, Cates K, Kerr C, Bermudez A, Susanto 
            TT, Wysong K, Garcia Marques FJ, Nolan GP, Pitteri S, Barna M
  TITLE     A subcellular map of translational machinery composition and 
            regulation at the single-molecule level.
  JOURNAL   Science. 2025 Mar 7;387(6738):eadn2623. doi: 
            10.1126/science.adn2623. Epub 2025 Mar 7.
  PUBMED    40048539
REFERENCE   2  (bases 1 to 6254)
  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: "Science."; 
            date: "2025-03-7"; volume: "387(6738):eadn2623. doi"; pages: " 
            10.1126/science.adn2623. Epub 2025 Mar 7"
COMMENT     Sequence Label: pCAGEN_NES_V5_eMagAF_C74_MAVS
FEATURES             Location/Qualifiers
     source          1..6254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        260..537
                     /label=chicken beta-actin promoter
     intron          538..1554
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1478..1500
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1562..1581
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1656..1697
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3150..3169)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3215..3270
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3269..3288)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3631..3647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3631..3647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3644..3666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3655..3671
                     /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(3679..3685,3686..3703,3704..3709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3724..3745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3804..3999
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3850..3985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4005..4139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4042..4061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4096..4115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4207..4224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4378..4966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4458..4477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5137..5928,5929..5997))
                     /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     5760..5779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5998..6102)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6133..6254,1..258)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
       61 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      121 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      181 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      241 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
      301 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
      361 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
      421 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      481 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      541 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      601 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      661 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      721 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
      781 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      841 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
      901 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
      961 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1021 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1081 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1141 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1201 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1261 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1321 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1381 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1441 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1501 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1561 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct cgaatgctgc
     1621 agaagaagct ggaggagctg gaactgggct cttccggtaa gcctatccct aaccctctcc
     1681 tcggtctcga ttctacggga tcctctggac acactcttta cgcccctgga ggatacgaca
     1741 ttatgggata tttggatcag attgcgaacc gcccaaaccc tcaggtcgaa ctggggcctg
     1801 tggacctgtc atgtgccctg atcctgtgcg atctgaagca aaaggacact ccgatcgtct
     1861 acgcctcgga agccttcttg gagatgaccg gatacaacag acatgaggtg ctcggcagga
     1921 actgcagatt cctgcagtcc cccgacggga tggtgaaacc aaagtcgact cgcaaatatg
     1981 tggactcgaa cacgatcttc accatcaaga aggccatcga ccggaacgcc gaggtccagg
     2041 tggaggtggt caactttaag aagaacggcc agcggttcgt gaactttctg accatcattc
     2101 cggtccggga tgaaaccgga gagtacagat actccatcgg attccagtgc gaaaccgaag
     2161 gcagtggatc cggcggcagt ggatccggcg gacaactgga cggtgggagt gttgccgtcc
     2221 tccctgtgat cgactccacg aaccagtact tgctggaccg catcggcgag cttaaaagcg
     2281 gagatgcttg tatcgctgaa taccagcagg ccgggcgcgg cagacggggc cggaagtggt
     2341 tctctccttt tggcgcaaat ctgtaccttt ctatgttctg gaggctggaa cagggaccag
     2401 cagccgcaat cggcttgtcc ctggtcattg ggattgtgat ggccgaagtt ctcagaaagc
     2461 tgggagctga taaggtgagg gttaaatggc ctaacgatct gtatttgcag gaccggaaac
     2521 tggccggaat actggtggag ctgactggca agactggcga cgccgctcag atagtcatcg
     2581 gagccggtat aaatatggcc atgaggcgcg tcgaggaatc cgtggtgaac caaggctgga
     2641 tcaccttgca agaggccggt attaacctgg atagaaacac cctggccgct atgctgatac
     2701 gcgaactgcg cgccgccctg gaactgttcg agcaggaggg actcgctcca tatctgagtc
     2761 gctgggaaaa attggataac ttcataaacc gacccgtgaa gctgatcatc ggcgacaagg
     2821 aaattttcgg aatctctcgc ggcatcgata aacagggggc attgctgctc gagcaggatg
     2881 ggattattaa gccatggatg ggcggggaaa tctcccttag atctgcagag aagggcagta
     2941 gcggatcctc tggctcttcc ccctcacctg gggctctgtg gctccaggtg gctgtgacag
     3001 gggtgctggt agtcacactc ctggtggtgc tgtaccggcg gcgtctgcac tgatcgagga
     3061 tatcgcggcc gcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     3121 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     3181 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     3241 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     3301 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     3361 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     3421 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     3481 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     3541 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     3601 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3781 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     3841 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3901 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3961 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     4021 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4081 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     4141 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4201 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4561 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4801 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5581 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5641 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5701 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5761 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5821 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5881 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5941 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6001 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6061 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6121 gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     6181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6241 gctgaccgcc caac
//
