LOCUS       sequence-467530-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi C-terminal fragment, residues 168-321, 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 5832)
  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 5832)
  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_C168
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..640
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          641..1657
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1581..1603
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1665..1684
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1759..1800
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2854..2873)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2919..2974
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2973..2992)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3335..3351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3335..3351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3348..3370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3359..3375
                     /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(3383..3389,3390..3407,3408..3413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3428..3449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3508..3703
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3554..3689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3709..3843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3746..3765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3800..3819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3911..3928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4082..4670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4162..4181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4841..5632,5633..5701))
                     /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     5464..5483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5702..5806)
                     /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 cggggggcgg ggcggggcga ggcggagagg
      541 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      601 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      661 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      721 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
      781 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
      841 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
      901 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
      961 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
     1021 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
     1081 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
     1141 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
     1201 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
     1261 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
     1321 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
     1381 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     1441 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     1501 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     1561 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1621 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1681 tgtgctgtct catcattttg gcaaagaatt cctcgaatgc tgcagaagaa gctggaggag
     1741 ctggaactgg gctcttccgg taagcctatc cctaaccctc tcctcggtct cgattctacg
     1801 ggatcctctg gacacactct ttacgcccct ggaggatacg acattatggg atatttggat
     1861 cagattgcga accgcccaaa ccctcaggtc gaactggggc ctgtggacct gtcatgtgcc
     1921 ctgatcctgt gcgatctgaa gcaaaaggac actccgatcg tctacgcctc ggaagccttc
     1981 ttggagatga ccggatacaa cagacatgag gtgctcggca ggaactgcag attcctgcag
     2041 tcccccgacg ggatggtgaa accaaagtcg actcgcaaat atgtggactc gaacacgatc
     2101 ttcaccatca agaaggccat cgaccggaac gccgaggtcc aggtggaggt ggtcaacttt
     2161 aagaagaacg gccagcggtt cgtgaacttt ctgaccatca ttccggtccg ggatgaaacc
     2221 ggagagtaca gatactccat cggattccag tgcgaaaccg aaggcagtgg atccggcggc
     2281 agtggatccg gcaaggtgag ggttaaatgg cctaacgatc tgtatttgca ggaccggaaa
     2341 ctggccggaa tactggtgga gctgactggc aagactggcg acgccgctca gatagtcatc
     2401 ggagccggta taaatatggc catgaggcgc gtcgaggaat ccgtggtgaa ccaaggctgg
     2461 atcaccttgc aagaggccgg tattaacctg gatagaaaca ccctggccgc tatgctgata
     2521 cgcgaactgc gcgccgccct ggaactgttc gagcaggagg gactcgctcc atatctgagt
     2581 cgctgggaaa aattggataa cttcataaac cgacccgtga agctgatcat cggcgacaag
     2641 gaaattttcg gaatctctcg cggcatcgat aaacaggggg cattgctgct cgagcaggat
     2701 gggattatta agccatggat gggcggggaa atctccctta gatctgcaga gaagtgatcg
     2761 aggatatcgc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     2821 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     2881 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     2941 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3001 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3061 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3121 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3181 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3241 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     3301 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3481 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     3721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3841 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4501 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4741 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4801 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4921 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4981 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5041 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5101 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5161 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5221 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5281 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5341 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5401 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5461 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5521 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5581 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5641 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5701 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5761 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5821 aagtgccacc tg
//
