LOCUS       sequence-467501-        5972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes ALIBi N-terminal fragment, residues 1-195, 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 5972)
  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 5972)
  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_N195_eMagBF_V5_NES
FEATURES             Location/Qualifiers
     source          1..5972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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             2814..2855
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2994..3013)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3059..3114
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3113..3132)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3475..3491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3475..3491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3488..3510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3499..3515
                     /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(3523..3529,3530..3547,3548..3553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3568..3589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3648..3843
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3694..3829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3849..3983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3886..3905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3940..3959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4051..4068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4222..4810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4302..4321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4981..5772,5773..5841))
                     /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     5604..5623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5842..5946)
                     /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 gccrgggggg gggggggggg gggrggggsg
      541 gggsggggsg rggsggrgag 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 gkgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg 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 tctcgaatga
     1741 aggacaacac tgttccgctt aagctgatag ccctcctggc caatggagag ttccacagcg
     1801 gtgagcagct cggtgagact ctcgggatgt ctcgcgccgc tattaacaag cacattcaga
     1861 cgttgcgcga ttggggagtc gacgtgttca cagtacctgg caaagggtac tcactccctg
     1921 agcctatcca gttgctgaac gcaaagcaga ttctgggaca actggacggt gggagtgttg
     1981 ccgtcctccc tgtgatcgac tccacgaacc agtacttgct ggaccgcatc ggcgagctta
     2041 aaagcggaga tgcttgtatc gctgaatacc agcaggccgg gcgcggcaga cggggccgga
     2101 agtggttctc tccttttggc gcaaatctgt acctttctat gttctggagg ctggaacagg
     2161 gaccagcagc cgcaatcggc ttgtccctgg tcattgggat tgtgatggcc gaagttctca
     2221 gaaagctggg agctgataag gtgagggtta aatggcctaa cgatctgtat ttgcaggacc
     2281 ggaaactggc cggaatactg gtggagctga ctggcaagac tggcagtgga tccggcggca
     2341 gtggatccgg cggacatacc ctctacgcgc cggggggtta tgacatcatg ggttacctca
     2401 gacagatcag aaaccggccg aacccacaag tggagctggg acccgtcgac ctctcctgcg
     2461 ccctcgtgct gtgtgacctt aagcagaagg acacccctgt ggtgtacgcc tccgaagcat
     2521 tcctggagat gaccgggtac aacagacacg aagtgctggg acggaactgc cgcttcctgc
     2581 aatccccgga tggaatggtg aagcctaagt caacccgcaa atacgtggac tccaacacta
     2641 tcttcaccat gaagaaggcc attgaccgca atgctgaggt gcaagtggaa gtggtgaact
     2701 tcaagaagaa cggacagcgc ttcgtcaact tcctgactat gattcccgtg cgggacgaaa
     2761 ccggcgaata ccggtacagc atcgggtttc agtgcgagac tgagggatcc tctggtaagc
     2821 ctatccctaa ccctctcctc ggtctcgatt ctacgggcag tagcctgcag aagaagctgg
     2881 aggagctgga actgtgatcg aggatatcgc ggccgcactc ctcaggtgca ggctgcctat
     2941 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3001 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3061 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3121 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3181 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     3241 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     3301 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     3361 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     3421 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     3481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     3661 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3781 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3841 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3901 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3961 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     4021 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4081 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4141 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4201 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4261 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4321 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4381 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4441 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4501 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4561 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4621 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4681 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4741 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4921 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4981 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5041 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5101 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5161 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5221 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5281 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5341 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5401 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5461 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5521 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5581 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5641 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5701 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5761 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5821 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5881 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5941 tccgcgcaca tttccccgaa aagtgccacc tg
//
