LOCUS       sequence-467531-        7082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF 
            (enhanced Magnet, acidic, fast), V5, Fbl for targeting to nucleolus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  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 7082)
  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_V5_Fbl_eMagAF_C74
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..642
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          643..1659
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1583..1605
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1667..1686
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1722..1763
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2562..2573)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     primer_bind     complement(4104..4123)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4169..4224
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4223..4242)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4585..4601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4585..4601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4598..4620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4609..4625
                     /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(4633..4639,4640..4657,4658..4663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4678..4699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4758..4953
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4804..4939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4959..5093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4996..5015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5050..5069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5161..5178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5332..5920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5412..5431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6091..6882,6883..6951))
                     /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     6714..6733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6952..7056)
                     /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 gggggggggg ggcggggggg ggggcggggc gaggcggaga
      541 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      601 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      661 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      721 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      781 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      841 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
      901 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
      961 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     1021 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     1081 gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct
     1141 gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg
     1201 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
     1261 gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
     1321 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
     1381 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     1441 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
     1501 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     1561 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     1621 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     1681 attgtgctgt ctcatcattt tggcaaagaa ttcctcgaat gggtaagcct atccctaacc
     1741 ctctcctcgg tctcgattct acgggatcct ctaagccagg tttcagcccc cgtgggggtg
     1801 gctttggtgg cagaggcggc tttggtgaca gaggcggtag aggaggtgga agaggaggcc
     1861 gaggaggctt tggcggtgga cgaggaggct ttggcggtgg aggtcgaggt cgaggcggag
     1921 ggggtggtgg cttcagggga cgaggaggtg gcggtggccg aggtgggggc ttccagtctg
     1981 ggggcaaccg gggtcgaggt ggtggccggg gaggcaagag aggaaaccag tcagggaaga
     2041 atgtgatggt ggagccgcat cgtcatgaag gtgtctttat ctgtcgcgga aaggaggatg
     2101 cccttgtcac aaagaatctg gtccctggag agtctgtgta tggagagaag agagtctcta
     2161 tttcagaagg agatgacaaa attgagtaca gagcctggaa ccccttccgc tccaagctgg
     2221 cagcagctat cctgggcggc gtagaccaga tccacatcaa gccaggagcc aaggtgctct
     2281 accttggggc agcctcaggc accaccgtct cccacgtctc tgatattgtc ggcccggatg
     2341 gtctggtcta cgcagttgag ttctcccacc gctctggccg tgacctcatc aacttggcca
     2401 agaagaggac taacattatt cctgtaattg aagatgctcg acacccacac aaataccgca
     2461 tgcttatcgc aatggtggat gtcatctttg ccgatgtggc ccagccagac caaacccgaa
     2521 ttgtggccct gaacgcccac accttcctgc ggaatggagg acactttgtg atttccatta
     2581 aggccaactg cattgactcc actgcgtcag cagaggctgt gtttgcatct gaagtgaaga
     2641 agatgcagca ggagaacatg aagccgcagg agcagctgac gctagagcct tatgagcgag
     2701 accacgccgt ggttgtcggt gtgtacaggc cacctcccaa ggtgaagaac ggcagtagcg
     2761 gatcctctgg ctcttccgga cacactcttt acgcccctgg aggatacgac attatgggat
     2821 atttggatca gattgcgaac cgcccaaacc ctcaggtcga actggggcct gtggacctgt
     2881 catgtgccct gatcctgtgc gatctgaagc aaaaggacac tccgatcgtc tacgcctcgg
     2941 aagccttctt ggagatgacc ggatacaaca gacatgaggt gctcggcagg aactgcagat
     3001 tcctgcagtc ccccgacggg atggtgaaac caaagtcgac tcgcaaatat gtggactcga
     3061 acacgatctt caccatcaag aaggccatcg accggaacgc cgaggtccag gtggaggtgg
     3121 tcaactttaa gaagaacggc cagcggttcg tgaactttct gaccatcatt ccggtccggg
     3181 atgaaaccgg agagtacaga tactccatcg gattccagtg cgaaaccgaa ggcagtggat
     3241 ccggcggcag tggatccggc ggacaactgg acggtgggag tgttgccgtc ctccctgtga
     3301 tcgactccac gaaccagtac ttgctggacc gcatcggcga gcttaaaagc ggagatgctt
     3361 gtatcgctga ataccagcag gccgggcgcg gcagacgggg ccggaagtgg ttctctcctt
     3421 ttggcgcaaa tctgtacctt tctatgttct ggaggctgga acagggacca gcagccgcaa
     3481 tcggcttgtc cctggtcatt gggattgtga tggccgaagt tctcagaaag ctgggagctg
     3541 ataaggtgag ggttaaatgg cctaacgatc tgtatttgca ggaccggaaa ctggccggaa
     3601 tactggtgga gctgactggc aagactggcg acgccgctca gatagtcatc ggagccggta
     3661 taaatatggc catgaggcgc gtcgaggaat ccgtggtgaa ccaaggctgg atcaccttgc
     3721 aagaggccgg tattaacctg gatagaaaca ccctggccgc tatgctgata cgcgaactgc
     3781 gcgccgccct ggaactgttc gagcaggagg gactcgctcc atatctgagt cgctgggaaa
     3841 aattggataa cttcataaac cgacccgtga agctgatcat cggcgacaag gaaattttcg
     3901 gaatctctcg cggcatcgat aaacaggggg cattgctgct cgagcaggat gggattatta
     3961 agccatggat gggcggggaa atctccctta gatctgcaga gaagtgatcg aggatatcgc
     4021 ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     4081 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     4141 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     4201 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     4261 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     4321 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     4381 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     4441 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     4501 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     4561 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4621 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4681 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4741 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4801 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4861 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4921 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     4981 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5041 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     5101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7081 tg
//
