LOCUS       sequence-468094-        7997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively encodes RPS2-smGFP-V5 (sfGFP-based spaghetti monster 
            fluorescent protein embedding V5 epitope tags), for targeting the 
            40S subunit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7997)
  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 7997)
  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: pBX_RPS2_smGFP_V5
FEATURES             Location/Qualifiers
     source          1..7997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1797..2813
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2737..2759
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2821..2840
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3812..3853
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3863..3904
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4436..4477
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4487..4528
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4547..4588
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4598..4639
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4853..4894
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4904..4945
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4955..4996
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5039..5056)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5045..5269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5316..5332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5340..5356
                     /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(5364..5370,5371..5388,5389..5394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5409..5430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5696..5758
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5809..5829)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5809..5827)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5848..5864)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5872..5888
                     /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(5896..5902,5903..5920,5921..5926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5941..5962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6079..6096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6250..6838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6330..6349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7009..7800,7801..7869))
                     /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     7632..7651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7870..7974)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     1441 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     1501 gggggggggg ggcgcgcgcc gggggggggg gggggggggg gggggggggg gggggggggg
     1561 gggggggggg gggggggggc ccaggggggg ggggggggcc ccaaagtttt ttttttggcg
     1621 gggggggggg gggggggggc cccctaaaaa agaagccggc ggcggggggg ggggggcgcg
     1681 gggcgccgcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
     1741 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
     1801 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
     1861 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     1921 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
     1981 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
     2041 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
     2101 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
     2161 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     2221 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
     2281 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
     2341 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     2401 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
     2461 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     2521 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     2581 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     2641 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     2701 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     2761 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     2821 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcaca ccatggcgga
     2881 tgacgccggt gcagcgggag ggcccggagg acccgggggc ccaggattag gtggccgcgg
     2941 cggcttccgc ggaggattcg gcagcggtct taggggccgt ggtcgtggcc gaggccgtgg
     3001 ccgtggtcga ggccgcgggg ctcgtggagg taaagctgaa gacaaggagt ggatccccgt
     3061 caccaagctg ggccgtctgg ttaaggacat gaagatcaag tccttggagg agatctacct
     3121 gttctccctg cccattaagg agtctgagat cattgatttc ttcctgggtg cgtccctaaa
     3181 ggatgaggtt ctgaaaatca tgccagtgca gaagcagact cgggctggcc agcggaccag
     3241 gttcaaggct ttcgtcgcta ttggggacta caatggtcac gttggtcttg gtgttaagtg
     3301 ctccaaggag gttgccactg ccatccgagg ggccatcatc ttggccaagc tttccatcgt
     3361 ccctgtgcgg agaggctact gggggaacaa gattggcaag ccccacactg ttccatgcaa
     3421 ggtgacaggc cgctgtggct ctgtgctggt gcgtctcatc cctgccccca gaggcactgg
     3481 cattgtctct gctcctgtgc ccaagaagct cctgatgatg gccggtatag atgactgcta
     3541 cacttcagcc agaggctgca ctgccaccct gggcaacttt gctaaggcca cctttgatgc
     3601 catctccaag acttacagct acctgactcc cgacctctgg aaagagactg tcttcaccaa
     3661 gtctccttat caggaattca cggatcatct tgtgaaaacc cacaccagag tctctgttca
     3721 gaggacccag gctccagctg tggctaccac aggatcgggc ggtaagccta tccaaaatcc
     3781 actgttgggg ctggatagca ctcaacagca aggaaaacca atacccaatc cactactggg
     3841 ccttgattca acgcagcagc agggtaaacc gatcccgaat cctcttcttg gcctcgatag
     3901 caccggtcaa ggggttagta agggagaaga actcttcaca ggtgttgtcc cgatacttgt
     3961 ggaactcgat ggggacgtca atggccataa gtttagcgtg agaggtgagg gcgagggcga
     4021 cgccacgaac ggtaagctga cacttaaatt tatatgcact acaggaaaac tccccgtgcc
     4081 ttggcccact ctcgtgacaa ccctcactta cggcgttcaa tgtttttcaa gatatcctga
     4141 tcacatgaag cagcatgact tttttaaatc cgcgatgccc gaaggatacg tccaagaacg
     4201 aactataagt tttaaggacg atggtactta taaaactcgc gcagaggtaa agttcgaagg
     4261 ggatactctt gttaacagga ttgaactcaa aggaatcgat ttcaaagagg acgggaatat
     4321 tcttgggcac aagctggagt acaactttaa cagccacaac gtgtacatca ccgctgacaa
     4381 gcaaaaaaat ggtatcaagg cgaactttaa gatacggcac aatgtagagg gtggcggaaa
     4441 gcccatccct aatccgctgc tgggtctgga tagtacgcag caacaaggta agccaatccc
     4501 taacccattg ctcggactcg actctaccgg tgggcaacaa ggtggtggaa agcctattcc
     4561 aaacccgctg cttggactgg atagcacgca acagcagggg aagccgattc ctaatcctct
     4621 cctggggttg gactccacag gcggggatgg aagtgtgcag cttgcggacc attatcagca
     4681 gaacactcca attggggacg gccccgtatt gcttcctgac aatcattatc tcagtactca
     4741 gagtgttctc tctaaagacc ccaatgagaa gcgagatcac atggtgctcc ttgagtttgt
     4801 gaccgccgca ggaattactc ttggaatgga cgagttgtat aaggggcaag gaggcaagcc
     4861 aatacctaat cccctcctgg ggctcgattc tacgcagcag caaggtaaac ccatacccaa
     4921 ccctcttttg ggactggact caactcagca gcaaggaaaa cctataccga acccgttgct
     4981 gggtctcgat tctacttgaa aacgatgatg catcgatgat ctagagctcg ctgatcagcc
     5041 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     5101 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     5161 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     5221 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     5281 gaaagaacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5341 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5401 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5461 tcgggaaacc tgtcgtgcca gcggatccat tcatgaatga attcatgtcg acatactagt
     5521 taaaagtttt gttactttat agaagaaatt ttgagttttt gttttttttt aataaataaa
     5581 taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa atataataaa
     5641 acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata aaacacatgc
     5701 gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt
     5761 aatctagtat acgcgtatgc ggccgcttaa ttaatccagc ttttgttccc tttagtgagg
     5821 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5881 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5941 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6661 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7981 tccccgaaaa gtgccac
//
