LOCUS       sequence-468093-        7681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively encodes RPL7a-smRuby2-HA (mRuby2-based spaghetti 
            monster fluorescent protein embedding HA epitope tags), for 
            targeting the 60S subunit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  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 7681)
  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_RPL7a_smRuby2_HA
FEATURES             Location/Qualifiers
     source          1..7681
                     /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"
     gap             1664..1673
                     /estimated_length=10
     intron          1799..2815
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2739..2761
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2823..2842
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3685..3711
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3715..3741
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3745..3771
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4279..4305
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4312..4338
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4351..4377
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4594..4620
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4624..4650
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4654..4680
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4723..4740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4729..4953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5000..5016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5024..5040
                     /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(5048..5054,5055..5072,5073..5078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5093..5114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5380..5442
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5493..5513)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5493..5511)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5532..5548)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5556..5572
                     /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(5580..5586,5587..5604,5605..5610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5625..5646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5763..5780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5934..6522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6014..6033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6693..7484,7485..7553))
                     /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     7316..7335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7554..7658)
                     /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 gggcggcccc gggggggggg gggggggggg gggggggggg gggggggggg
     1561 gggggggggg gggggggggc gcgcccaggg gggggggggg ggggcggctt cttttttttg
     1621 cggcggcgcg ggggcggggg gcgccgcaaa aaggcgagtg cggnnnnnnn nnnggcgggg
     1681 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     1741 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     1801 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     1861 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     1921 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
     1981 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
     2041 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
     2101 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     2161 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     2221 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     2281 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     2341 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     2401 gccgcctcgg gccggggggg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     2461 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     2521 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     2581 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     2641 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     2701 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     2761 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     2821 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattca caccatgccc
     2881 aaagggaaga aggccaaggg gaagaaggtg gccccggccc ccgccgtcgt caagaagcag
     2941 gaggccaaga aggtggtcaa ccctctgttc gagaagaggc ccaagaactt cggcattgga
     3001 caggacatcc agcccaaaag agatttaacg cgcttcgtca aatggccccg ctacatcaga
     3061 ctgcagcggc aaagagctat cctctacaag cggctcaaag tccctcctgc cattaaccag
     3121 ttcacccagg ccctggacag gcagacagct actcagctgc ttaagcttgc ccacaagtac
     3181 aggccagaga caaagcaaga gaagaagcaa aggctactgg cccgtgctga gaagaaagct
     3241 gctggcaaag gcgacgtccc aactaagaga ccacctgtcc tccgagcagg agtcaataca
     3301 gtcaccacct tggtggagaa caagaaggct cagctggtgg tgattgccca tgacgtagac
     3361 cccattgagc tggtggtttt cctacctgct ctgtgtcgaa agatgggggt gccctactgc
     3421 atcatcaagg gaaaggccag gctggggcac ctggtccaca ggaagacatg caccaccgtt
     3481 gccttcacac aggttaactc ggaagacaag ggtgctctgg ctaagctggt ggaagctatt
     3541 aggaccaatt ataatgacag atatgacgag atccgtcgcc actggggagg caacgtcctg
     3601 ggtcctaagt ctgtggctcg aattgccaag ctggaaaaag caaaggctaa agaactcgcc
     3661 actaaattgg gtggatcggg cgtttatccc tatgatgtgc cggattatgc tggataccca
     3721 tacgatgtgc ctgattatgc aggatatccg tacgatgtgc cagattacgc gggtggaaat
     3781 tctcttataa aagaaaatat gaggatgaag gtggttctgg agggctcagt caatgggcac
     3841 cagttcaaat gcactggaga aggagaaggt aatccttata tgggcaccca aacaatgagg
     3901 atcaaggtta ttgaaggtgg accgttgccc tttgctttcg atatcctggc gacgagtttt
     3961 atgtacggaa gtcggacttt tataaagtat ccaaaaggca ttcctgattt tttcaagcaa
     4021 tcctttccgg aaggctttac atgggagaga gtgacccggt acgaggatgg cggtgtgatc
     4081 acagtcatgc aagatacgag cttggaagac ggatgcttgg tgtaccacgc ccaagtacga
     4141 ggagttaatt tcccatctaa cggcgccgtc atgcaaaaga aaactaaagg atgggagccc
     4201 aacactgaga tgatgtatcc tgcagatggc ggtttgagag gctatactca catggcgctt
     4261 aaagtagacg gcggaggtta tccttatgat gtacctgatt atgctggtgg ctacccttac
     4321 gatgtaccag actacgccgg tggcggcggt tatccgtacg acgtgccaga ctacgcagga
     4381 gggcaccctt atgacgtgcc cgactacgcg ggaggacacc tttcatgttc tttcgttacc
     4441 acttatcgct ctaagaagac cgtgggcaat ataaaaatgc ccggaataca tgcagtggac
     4501 cataggctcg aacggctcga agaatctgat aacgaaatgt ttgttgttca acgcgaacac
     4561 gccgtggcta agttcgcagg tctcggaggc ggctatcctt acgacgtccc tgactacgca
     4621 ggatacccgt acgatgtgcc cgattacgca ggttacccgt atgatgtacc ggactatgcg
     4681 taaaaacgat gatgcatcga tgatctagag ctcgctgatc agcctcgact gtgccttcta
     4741 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4801 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4861 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4921 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga acctgcagcc
     4981 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5161 gccagcggat ccattcatga atgaattcat gtcgacatac tagttaaaag ttttgttact
     5221 ttatagaaga aattttgagt ttttgttttt ttttaataaa taaataaaca taaataaatt
     5281 gtttgttgaa tttattatta gtatgtaagt gtaaatataa taaaacttaa tatctattca
     5341 aattaataaa taaacctcga tatacagacc gataaaacac atgcgtcaat tttacgcatg
     5401 attatcttta acgtacgtca caatatgatt atctttctag ggttaatcta gtatacgcgt
     5461 atgcggccgc ttaattaatc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     5521 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5581 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5641 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7681 c
//
