LOCUS       sequence-429917-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP P2A RFP control reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Inglis AJ, Guna A, Galvez-Merchan A, Pal A, Esantsi TK, Keys HR, 
            Frenkel EM, Oania R, Weissman JS, Voorhees RM
  TITLE     Coupled protein quality control during nonsense-mediated mRNA decay.
  JOURNAL   J Cell Sci. 2023 May 15;136(10):jcs261216. doi: 10.1242/jcs.261216. 
            Epub 2023 May 23.
  PUBMED    37218462
REFERENCE   2  (bases 1 to 6536)
  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: "J Cell 
            Sci."; date: "2023-05-15"; volume: "136(10):jcs261216. doi"; pages: 
            " 10.1242/jcs.261216. Epub 2023 May 23"
COMMENT     Sequence Label: GPR_control
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..88)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        260..639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        640..843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     794..814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    845..863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    866..884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     894..918
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1731..1787
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2513..2530)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2519..2743
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2876..2895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3027..3074
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3082..4104
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3082..4104
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4234..4355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4271..4290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4325..4344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4404..4420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4404..4420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4417..4439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4428..4444
                     /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(4452..4458,4459..4476,4477..4482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4497..4518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4635..4652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4806..5394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4886..4905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5565..6356,6357..6425))
                     /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     6188..6207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6426..6530)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
       61 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
      121 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
      181 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
      241 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
      301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      601 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      661 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      721 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      781 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      841 gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt
      901 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      961 caccgggacc gatccagcct ccggactcta gcgtttaaac ttaagcttac ggtaccatgg
     1021 tgtctaaagg tgaagaactt tttacgggtg tcgtccccat cctggttgaa ttggacggag
     1081 acgtgaatgg ccataaattt tcagtgagcg gtgaaggaga gggagacgcc acttacggta
     1141 aacttacctt gaaatttatc tgcactaccg ggaaactgcc cgtaccctgg ccgacgctcg
     1201 taacaacgct tacctacggc gtgcaatgtt ttagcagata cccagaccat atgaagcagc
     1261 acgatttttt caagagtgca atgcctgagg ggtacgtaca ggaaaggacg atctttttca
     1321 aagacgatgg taattataaa acgcgggctg aggttaaatt tgaaggagac acgctcgtga
     1381 atcgaataga gctgaaggga atcgacttca aggaagacgg taacatcctg ggccataaac
     1441 tcgaatataa ttacaattca cacaacgtct atatcatggc agataaacag aagaatggca
     1501 tcaaagtcaa ttttaagatc cggcataata tcgaggacgg atcagtacag cttgccgacc
     1561 attaccaaca gaatacaccg ataggagacg gtcccgtcct cttgccagat aatcactatt
     1621 tgagcacaca atccgctctt tctaaagatc cgaatgaaaa gcgcgatcat atggtcttga
     1681 aagagttcgt cactgccgcc ggcataaccg gcggatccgc cggaagcgga gctactaact
     1741 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctggc accggttcca
     1801 acatggcgat tatcaaggaa ttcatgaggt tcaaagttca catggaggga tctgtaaacg
     1861 gccacgagtt cgaaatcgag ggagaggggg agggtcgacc atatgaagga acacagaccg
     1921 ctaaattgaa ggttactaaa ggtggaccac ttcctttcgc atgggacatt cttagccccc
     1981 agtttatgta tggttccaag gcgtatgtca aacatcccgc agacattcct gattacctta
     2041 aactgagttt tccagagggg ttcaaatggg agcgcgtcat gaactttgaa gatgggggcg
     2101 ttgttacagt gactcaggac tcttctttgc aagatggaga gtttatatat aaggtgaaac
     2161 ttcgcggaac caacttccca agcgatggcc cagtcatgca gaagaaaact atgggttggg
     2221 aagccagtag cgagcggatg tacccagagg acggcgcact taagggtgag attaagcagc
     2281 ggctgaaact caaagacgga ggccattatg acgcggaagt taaaacgacg tacaaagcta
     2341 aaaagcctgt ccaattgcca ggcgcgtaca atgtcaatat taaattggac attaccagtc
     2401 acaacgaaga ttacacaata gtggagcaat acgaacgcgc agagggacgc cattcaacag
     2461 ggtaagcggc cgctactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2521 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2581 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2641 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2701 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2761 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2821 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2881 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2941 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3001 gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa
     3061 agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt
     3121 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     3181 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     3241 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     3301 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt
     3361 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     3421 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     3481 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     3541 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     3601 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     3661 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     3721 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     3781 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     3841 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     3901 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     3961 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     4021 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     4081 actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc
     4141 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4201 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4261 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4321 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4381 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4441 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4501 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4561 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5701 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5761 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5821 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5881 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5941 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6001 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6061 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6121 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6181 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6241 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6301 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6361 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6421 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatt
//
