LOCUS       sequence-371400-        7217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses RPL13 with C-terminal tags ECFP and SNAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7217)
  AUTHORS   Vester K, Preussner M, Holton N, Feng S, Schultz C, Heyd F, Wahl MC
  TITLE     Recruitment of a splicing factor to the nuclear lamina for its 
            inactivation.
  JOURNAL   Commun Biol. 2022 Jul 22;5(1):736. doi: 10.1038/s42003-022-03689-y.
  PUBMED    35869234
REFERENCE   2  (bases 1 to 7217)
  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: "Commun 
            Biol."; date: "2022-07-22"; volume: "5(1):736. doi"; pages: " 
            10.1038/s42003-022-03689-y"
COMMENT     Sequence Label: RPL13-ECFP-SNAP
FEATURES             Location/Qualifiers
     source          1..7217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1563..1565,1566..1568,1569..2279)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1608..1629)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1869..1888)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2216..2237
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2337..2882
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          2993..3222
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3205..3224
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3295..3869,3870..3872,3873..3881)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3462..3479)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3689..3708
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3904..4707
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3967..3986)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4577..4596
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4837..4958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4874..4893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5060..5076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5060..5076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5073..5095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5084..5100
                     /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(5108..5114,5115..5132,5133..5138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5153..5174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5291..5308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5462..6050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5542..5561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6221..7012,7013..7081))
                     /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     6844..6863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7082..7186)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgtcgctagc accatggcgc ccagccggaa tggcatggtc ttgaagcccc
      961 acttccacaa ggactggcag cggcgcgtgg ccacgtggtt caaccagccg gcccgtaaga
     1021 tccgcagacg taaggcccgg caagccaagg cgcgccgcat cgccccgcgc cccgcgtcgg
     1081 gtcccatccg gcccatcgtg cgctgcccca cggttcggta ccacacgaag gtgcgcgccg
     1141 gccgcggctt cagcctggag gagctcaggg tggccggcat tcacaagaag gtggcccgga
     1201 ccatcggcat ttctgtggat ccgaggaggc ggaacaagtc cacggagtcc ctgcaggcca
     1261 acgtgcagcg gctgaaggag taccgctcca aactcatcct cttccccagg aagccctcgg
     1321 cccccaagaa gggagacagt tctgctgaag aactgaaact ggccacccag ctgaccggac
     1381 cggtcatgcc cgtccggaac gtctataaga aggagaaagc tcgagtcatc actgaggaag
     1441 agaagaattt caaagccttc gctagtctcc gtatggcccg tgccaacgcc cggctcttcg
     1501 gcatacgggc aaaaagagcc aaggaagccg cagaacagga tgttgaaaag aaaaaagcta
     1561 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1621 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1681 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1741 ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc gaccacatga
     1801 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1861 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1921 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1981 acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac aagcagaaga
     2041 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2101 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2161 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2221 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2281 ccggactcag atctcgagct caagcttcga attctgcagg taccggtgaa ttcaccatgg
     2341 acaaagactg cgaaatgaag cgcaccaccc tggatagccc tctgggcaag ctggaactgt
     2401 ctgggtgcga acagggcctg caccgtatca tcttcctggg caaaggaaca tctgccgccg
     2461 acgccgtgga agtgcctgcc ccagccgccg tgctgggcgg accagagcca ctgatgcagg
     2521 ccaccgcctg gctcaacgcc tactttcacc agcctgaggc catcgaggag ttccctgtgc
     2581 cagccctgca ccacccagtg ttccagcagg agagctttac ccgccaggtg ctgtggaaac
     2641 tgctgaaagt ggtgaagttc ggagaggtca tcagctacag ccacctggcc gccctggccg
     2701 gcaatcccgc cgccaccgcc gccgtgaaaa ccgccctgag cggaaatccc gtgcccattc
     2761 tgatcccctg ccaccgggtg gtgcagggcg acctggacgt ggggggctac gagggcgggc
     2821 tcgccgtgaa agagtggctg ctggcccacg agggccacag actgggcaag cctgggctgg
     2881 gtcctgcagg cggatccgcg tttaaactcg aggttaatta atgagcggcc gcatagataa
     2941 ctgatccagt gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac
     3001 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag
     3061 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca
     3121 gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac
     3181 acttgagtga caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc
     3241 aactgcaggt cgagcatgca tctagggcgg ccaattccgc ccctctcccc cccccccctc
     3301 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     3361 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     3421 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     3481 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     3541 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     3601 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     3661 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     3721 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     3781 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     3841 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggataat tcctgcagcc
     3901 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3961 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4021 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4081 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4141 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4201 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4261 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4321 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4381 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4441 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4501 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4561 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4621 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4681 tatcgccttc ttgacgagtt cttctgaggg gatcaattct ctagataact gatcataatc
     4741 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4801 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4861 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4921 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg cattctagtt
     4981 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5041 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5101 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5161 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5221 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5281 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5341 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5401 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5461 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5521 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5581 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5641 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5701 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5761 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5821 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5881 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5941 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6481 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6541 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6601 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6661 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6721 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6781 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6841 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6901 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6961 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7021 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7081 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7141 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7201 aagtgccacc tgacgtc
//
