LOCUS       sequence-467543-        6228 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, 
            MyrPalm for targeting to plasma membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6228)
  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 6228)
  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_MyrPalm_NES_V5_eMagAF_C74
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1873..1914
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3250..3269)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3315..3370
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3369..3388)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3731..3747)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3731..3747)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3744..3766)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3755..3771
                     /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(3779..3785,3786..3803,3804..3809))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3824..3845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3904..4099
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3950..4085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4105..4239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4142..4161)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4196..4215
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4307..4324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4478..5066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4558..4577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5237..6028,6029..6097))
                     /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     5860..5879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6098..6202)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgaatg
     1741 ggctgcatca agagcaagcg caaggacaac ctgaacgacg acggcgtgga catgaagacc
     1801 aagggcagtg gatccaccag tggcagtgga tccctgcaga agaagctgga ggagctggaa
     1861 ctgggctctt ccggtaagcc tatccctaac cctctcctcg gtctcgattc tacgggatcc
     1921 tctggacaca ctctttacgc ccctggagga tacgacatta tgggatattt ggatcagatt
     1981 gcgaaccgcc caaaccctca ggtcgaactg gggcctgtgg acctgtcatg tgccctgatc
     2041 ctgtgcgatc tgaagcaaaa ggacactccg atcgtctacg cctcggaagc cttcttggag
     2101 atgaccggat acaacagaca tgaggtgctc ggcaggaact gcagattcct gcagtccccc
     2161 gacgggatgg tgaaaccaaa gtcgactcgc aaatatgtgg actcgaacac gatcttcacc
     2221 atcaagaagg ccatcgaccg gaacgccgag gtccaggtgg aggtggtcaa ctttaagaag
     2281 aacggccagc ggttcgtgaa ctttctgacc atcattccgg tccgggatga aaccggagag
     2341 tacagatact ccatcggatt ccagtgcgaa accgaaggca gtggatccgg cggcagtgga
     2401 tccggcggac aactggacgg tgggagtgtt gccgtcctcc ctgtgatcga ctccacgaac
     2461 cagtacttgc tggaccgcat cggcgagctt aaaagcggag atgcttgtat cgctgaatac
     2521 cagcaggccg ggcgcggcag acggggccgg aagtggttct ctccttttgg cgcaaatctg
     2581 tacctttcta tgttctggag gctggaacag ggaccagcag ccgcaatcgg cttgtccctg
     2641 gtcattggga ttgtgatggc cgaagttctc agaaagctgg gagctgataa ggtgagggtt
     2701 aaatggccta acgatctgta tttgcaggac cggaaactgg ccggaatact ggtggagctg
     2761 actggcaaga ctggcgacgc cgctcagata gtcatcggag ccggtataaa tatggccatg
     2821 aggcgcgtcg aggaatccgt ggtgaaccaa ggctggatca ccttgcaaga ggccggtatt
     2881 aacctggata gaaacaccct ggccgctatg ctgatacgcg aactgcgcgc cgccctggaa
     2941 ctgttcgagc aggagggact cgctccatat ctgagtcgct gggaaaaatt ggataacttc
     3001 ataaaccgac ccgtgaagct gatcatcggc gacaaggaaa ttttcggaat ctctcgcggc
     3061 atcgataaac agggggcatt gctgctcgag caggatggga ttattaagcc atggatgggc
     3121 ggggaaatct cccttagatc tgcagagaag tgatcgagga tatcgcggcc gcactcctca
     3181 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     3241 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     3301 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3361 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     3421 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     3481 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     3541 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     3601 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     3661 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg
     3721 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3781 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3841 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     3901 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3961 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4021 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4081 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     4141 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4201 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     4261 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4321 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4381 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4441 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4501 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4561 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4621 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4681 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4741 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4801 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4861 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4921 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4981 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5041 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5101 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5161 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5221 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5281 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5341 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5401 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5461 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5521 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5581 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5641 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5701 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5761 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5821 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5881 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5941 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6001 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6061 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6121 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6181 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
