LOCUS       sequence-467516-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes full-length BirA fused to a nuclear export sequence and V5. 
            Used as a positive control in ALIBi experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  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 5855)
  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_BirA
FEATURES             Location/Qualifiers
     source          1..5855
                     /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             1776..1817
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1818..2777
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(2877..2896)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2942..2997
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2996..3015)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3358..3374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3358..3374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3371..3393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3382..3398
                     /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(3406..3412,3413..3430,3431..3436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3451..3472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3531..3726
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3577..3712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3732..3866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3769..3788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3823..3842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3934..3951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4105..4693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4185..4204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4864..5655,5656..5724))
                     /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     5487..5506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5725..5829)
                     /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 tccatgctgc
     1741 agaagaagct ggaggagctg gaactgggct cttccggtaa gcctatccct aaccctctcc
     1801 tcggtctcga ttctacgaag gacaacactg ttccgcttaa gctgatagcc ctcctggcca
     1861 atggagagtt ccacagcggt gagcagctcg gtgagactct cgggatgtct cgcgccgcta
     1921 ttaacaagca cattcagacg ttgcgcgatt ggggagtcga cgtgttcaca gtacctggca
     1981 aagggtactc actccctgag cctatccagt tgctgaacgc aaagcagatt ctgggacaac
     2041 tggacggtgg gagtgttgcc gtcctccctg tgatcgactc cacgaaccag tacttgctgg
     2101 accgcatcgg cgagcttaaa agcggagatg cttgtatcgc tgaataccag caggccgggc
     2161 gcggcagacg gggccggaag tggttctctc cttttggcgc aaatctgtac ctttctatgt
     2221 tctggaggct ggaacaggga ccagcagccg caatcggctt gtccctggtc attgggattg
     2281 tgatggccga agttctcaga aagctgggag ctgataaggt gagggttaaa tggcctaacg
     2341 atctgtattt gcaggaccgg aaactggccg gaatactggt ggagctgact ggcaagactg
     2401 gcgacgccgc tcagatagtc atcggagccg gtataaatat ggccatgagg cgcgtcgagg
     2461 aatccgtggt gaaccaaggc tggatcacct tgcaagaggc cggtattaac ctggatagaa
     2521 acaccctggc cgctatgctg atacgcgaac tgcgcgccgc cctggaactg ttcgagcagg
     2581 agggactcgc tccatatctg agtcgctggg aaaaattgga taacttcata aaccgacccg
     2641 tgaagctgat catcggcgac aaggaaattt tcggaatctc tcgcggcatc gataaacagg
     2701 gggcattgct gctcgagcag gatgggatta ttaagccatg gatgggcggg gaaatctccc
     2761 ttagatctgc agagaagtga tcgaggatat cgcggccgca ctcctcaggt gcaggctgcc
     2821 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     2881 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     2941 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3001 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3061 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     3121 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     3181 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     3241 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     3301 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     3361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     3541 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3601 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3661 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3721 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3781 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3841 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     3901 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3961 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4021 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4081 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4141 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4201 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4261 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4321 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4381 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4441 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4501 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4561 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4621 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4681 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4741 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4801 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4861 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4921 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4981 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5041 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5101 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5161 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5221 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5281 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5341 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5401 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5461 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5521 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5581 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5641 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5701 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5761 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5821 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
