LOCUS       sequence-467528-        6159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi C-terminal fragment, residues 66-321, fused to eMagAF 
            (enhanced Magnet, acidic, fast), V5, and nuclear export sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  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 6159)
  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_C66
FEATURES             Location/Qualifiers
     source          1..6159
                     /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             1780..1821
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3181..3200)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3246..3301
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3300..3319)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3662..3678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3662..3678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3675..3697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3686..3702
                     /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(3710..3716,3717..3734,3735..3740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3755..3776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3835..4030
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3881..4016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4036..4170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4073..4092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4127..4146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4238..4255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4409..4997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4489..4508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5168..5959,5960..6028))
                     /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     5791..5810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6029..6133)
                     /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 gggggggcgc 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 ggggcggcct 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 ctgcagaaga agctggagga gctggaactg ggctcttccg gtaagcctat ccctaaccct
     1801 ctcctcggtc tcgattctac gggatcctct ggacacactc tttacgcccc tggaggatac
     1861 gacattatgg gatatttgga tcagattgcg aaccgcccaa accctcaggt cgaactgggg
     1921 cctgtggacc tgtcatgtgc cctgatcctg tgcgatctga agcaaaagga cactccgatc
     1981 gtctacgcct cggaagcctt cttggagatg accggataca acagacatga ggtgctcggc
     2041 aggaactgca gattcctgca gtcccccgac gggatggtga aaccaaagtc gactcgcaaa
     2101 tatgtggact cgaacacgat cttcaccatc aagaaggcca tcgaccggaa cgccgaggtc
     2161 caggtggagg tggtcaactt taagaagaac ggccagcggt tcgtgaactt tctgaccatc
     2221 attccggtcc gggatgaaac cggagagtac agatactcca tcggattcca gtgcgaaacc
     2281 gaaggcagtg gatccggcgg cagtggatcc ggcttgctga acgcaaagca gattctggga
     2341 caactggacg gtgggagtgt tgccgtcctc cctgtgatcg actccacgaa ccagtacttg
     2401 ctggaccgca tcggcgagct taaaagcgga gatgcttgta tcgctgaata ccagcaggcc
     2461 gggcgcggca gacggggccg gaagtggttc tctccttttg gcgcaaatct gtacctttct
     2521 atgttctgga ggctggaaca gggaccagca gccgcaatcg gcttgtccct ggtcattggg
     2581 attgtgatgg ccgaagttct cagaaagctg ggagctgata aggtgagggt taaatggcct
     2641 aacgatctgt atttgcagga ccggaaactg gccggaatac tggtggagct gactggcaag
     2701 actggcgacg ccgctcagat agtcatcgga gccggtataa atatggccat gaggcgcgtc
     2761 gaggaatccg tggtgaacca aggctggatc accttgcaag aggccggtat taacctggat
     2821 agaaacaccc tggccgctat gctgatacgc gaactgcgcg ccgccctgga actgttcgag
     2881 caggagggac tcgctccata tctgagtcgc tgggaaaaat tggataactt cataaaccga
     2941 cccgtgaagc tgatcatcgg cgacaaggaa attttcggaa tctctcgcgg catcgataaa
     3001 cagggggcat tgctgctcga gcaggatggg attattaagc catggatggg cggggaaatc
     3061 tcccttagat ctgcagagaa gtgatcgagg atatcgcggc cgcactcctc aggtgcaggc
     3121 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     3181 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     3241 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     3301 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     3361 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     3421 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     3481 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     3541 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     3601 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     3661 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     3841 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3901 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3961 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4021 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4141 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     4201 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6061 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6121 taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
