LOCUS       sequence-467577-        6573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF 
            (enhanced Magnet, basic, fast), V5, Fbl for targeting to nucleolus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6573)
  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 6573)
  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_Fbl_N73_eMagBF_V5
FEATURES             Location/Qualifiers
     source          1..6573
                     /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             complement(2530..2541)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             3454..3495
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3595..3614)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3660..3715
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3714..3733)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4076..4092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4076..4092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4089..4111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4100..4116
                     /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(4124..4130,4131..4148,4149..4154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4169..4190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4249..4444
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4295..4430
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4450..4584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4487..4506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4541..4560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4652..4669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4823..5411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4903..4922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5582..6373,6374..6442))
                     /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     6205..6224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6443..6547)
                     /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 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 aagccaggtt tcagcccccg tgggggtggc tttggtggca gaggcggctt tggtgacaga
     1801 ggcggtagag gaggtggaag aggaggccga ggaggctttg gcggtggacg aggaggcttt
     1861 ggcggtggag gtcgaggtcg aggcggaggg ggtggtggct tcaggggacg aggaggtggc
     1921 ggtggccgag gtgggggctt ccagtctggg ggcaaccggg gtcgaggtgg tggccgggga
     1981 ggcaagagag gaaaccagtc agggaagaat gtgatggtgg agccgcatcg tcatgaaggt
     2041 gtctttatct gtcgcggaaa ggaggatgcc cttgtcacaa agaatctggt ccctggagag
     2101 tctgtgtatg gagagaagag agtctctatt tcagaaggag atgacaaaat tgagtacaga
     2161 gcctggaacc ccttccgctc caagctggca gcagctatcc tgggcggcgt agaccagatc
     2221 cacatcaagc caggagccaa ggtgctctac cttggggcag cctcaggcac caccgtctcc
     2281 cacgtctctg atattgtcgg cccggatggt ctggtctacg cagttgagtt ctcccaccgc
     2341 tctggccgtg acctcatcaa cttggccaag aagaggacta acattattcc tgtaattgaa
     2401 gatgctcgac acccacacaa ataccgcatg cttatcgcaa tggtggatgt catctttgcc
     2461 gatgtggccc agccagacca aacccgaatt gtggccctga acgcccacac cttcctgcgg
     2521 aatggaggac actttgtgat ttccattaag gccaactgca ttgactccac tgcgtcagca
     2581 gaggctgtgt ttgcatctga agtgaagaag atgcagcagg agaacatgaa gccgcaggag
     2641 cagctgacgc tagagcctta tgagcgagac cacgccgtgg ttgtcggtgt gtacaggcca
     2701 cctcccaagg tgaagaacgg cagtagcgga tcctctggct cttccaagga caacactgtt
     2761 ccgcttaagc tgatagccct cctggccaat ggagagttcc acagcggtga gcagctcggt
     2821 gagactctcg ggatgtctcg cgccgctatt aacaagcaca ttcagacgtt gcgcgattgg
     2881 ggagtcgacg tgttcacagt acctggcaaa gggtactcac tccctgagcc tatccagttg
     2941 ctgaacgcaa agcagattct gggcagtgga tccggcggca gtggatccgg cggacatacc
     3001 ctctacgcgc cggggggtta tgacatcatg ggttacctca gacagatcag aaaccggccg
     3061 aacccacaag tggagctggg acccgtcgac ctctcctgcg ccctcgtgct gtgtgacctt
     3121 aagcagaagg acacccctgt ggtgtacgcc tccgaagcat tcctggagat gaccgggtac
     3181 aacagacacg aagtgctggg acggaactgc cgcttcctgc aatccccgga tggaatggtg
     3241 aagcctaagt caacccgcaa atacgtggac tccaacacta tcttcaccat gaagaaggcc
     3301 attgaccgca atgctgaggt gcaagtggaa gtggtgaact tcaagaagaa cggacagcgc
     3361 ttcgtcaact tcctgactat gattcccgtg cgggacgaaa ccggcgaata ccggtacagc
     3421 atcgggtttc agtgcgagac tgagggatcc tctggtaagc ctatccctaa ccctctcctc
     3481 ggtctcgatt ctacgtgatc gaggatatcg cggccgcact cctcaggtgc aggctgccta
     3541 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3601 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3661 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3721 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3781 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3841 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3901 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     3961 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     4021 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     4081 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4141 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4201 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     4261 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4321 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4381 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4441 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4501 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4561 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     4621 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4681 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6541 ttccgcgcac atttccccga aaagtgccac ctg
//
