LOCUS       sequence-467537-        5809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi N-terminal fragment, residues 1-140, fused to eMagBF 
            (enhanced Magnet, basic, fast), V5, and nuclear export sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5809)
  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 5809)
  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_N140_eMagBF_V5_NES
FEATURES             Location/Qualifiers
     source          1..5809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2651..2692
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2831..2850)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2896..2951
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2950..2969)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3312..3328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3312..3328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3325..3347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3336..3352
                     /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(3360..3366,3367..3384,3385..3390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3405..3426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3485..3680
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3531..3666
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3686..3820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3723..3742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3777..3796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3888..3905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4059..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4139..4158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4818..5609,5610..5678))
                     /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     5441..5460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5679..5783)
                     /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 gcgggggggg cggggggggg ggggggcgcg cgccaggcgg gggggggggg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgaat
     1741 gaaggacaac actgttccgc ttaagctgat agccctcctg gccaatggag agttccacag
     1801 cggtgagcag ctcggtgaga ctctcgggat gtctcgcgcc gctattaaca agcacattca
     1861 gacgttgcgc gattggggag tcgacgtgtt cacagtacct ggcaaagggt actcactccc
     1921 tgagcctatc cagttgctga acgcaaagca gattctggga caactggacg gtgggagtgt
     1981 tgccgtcctc cctgtgatcg actccacgaa ccagtacttg ctggaccgca tcggcgagct
     2041 taaaagcgga gatgcttgta tcgctgaata ccagcaggcc gggcgcggca gacggggccg
     2101 gaagtggttc tctccttttg gcgcaaatct gtacctttct atgttctgga ggctggaagg
     2161 cagtggatcc ggcggcagtg gatccggcgg acataccctc tacgcgccgg ggggttatga
     2221 catcatgggt tacctcagac agatcagaaa ccggccgaac ccacaagtgg agctgggacc
     2281 cgtcgacctc tcctgcgccc tcgtgctgtg tgaccttaag cagaaggaca cccctgtggt
     2341 gtacgcctcc gaagcattcc tggagatgac cgggtacaac agacacgaag tgctgggacg
     2401 gaactgccgc ttcctgcaat ccccggatgg aatggtgaag cctaagtcaa cccgcaaata
     2461 cgtggactcc aacactatct tcaccatgaa gaaggccatt gaccgcaatg ctgaggtgca
     2521 agtggaagtg gtgaacttca agaagaacgg acagcgcttc gtcaacttcc tgactatgat
     2581 tcccgtgcgg gacgaaaccg gcgaataccg gtacagcatc gggtttcagt gcgagactga
     2641 gggatcctct ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgggcagtag
     2701 cctgcagaag aagctggagg agctggaact gtgatcgagg atatcgcggc cgcactcctc
     2761 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2821 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2881 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     2941 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3001 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     3061 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     3121 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3181 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3241 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     3301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     3481 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3541 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3601 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3661 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     3721 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3781 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
