LOCUS       sequence-423967-        6168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Blue Fluorescent Protein - P2A - short length 
            protein kinase R from chinook salmon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  AUTHORS   Chaumont L, Peruzzi M, Huetz F, Raffy C, Le Hir J, Minke J, Boudinot
            P, Collet B
  TITLE     Salmonid Double-stranded RNA-Dependent Protein Kinase Activates 
            Apoptosis and Inhibits Protein Synthesis.
  JOURNAL   J Immunol. 2024 Jul 26:ji2400076. doi: 10.4049/jimmunol.2400076.
  PUBMED    39058317
REFERENCE   2  (bases 1 to 6168)
  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 Immunol. 
            2024 Jul 26:ji2400076. doi: 10.4049/jimmunol.2400076."
COMMENT     Sequence Label: pcDNA3.1-Zeo-BFP-P2A-otPKR-SL
FEATURES             Location/Qualifiers
     source          1..6168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..84)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        256..635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        636..839
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     790..810
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     884..903
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        884..902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1650..1706
                     /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(2197..2214)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2203..2427
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2473..2901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2560..2579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2770..2791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2910..2930)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2915..3244
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3095..3230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3157..3176
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3292..3339
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3358..3732
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3862..3983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3899..3918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3953..3972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4032..4048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4032..4048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4045..4067)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4056..4072
                     /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(4080..4086,4087..4104,4105..4110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4125..4146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4263..4280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4434..5022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4514..4533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5193..5984,5985..6053))
                     /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     5816..5835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6054..6158)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
       61 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
      121 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
      181 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      241 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      301 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      361 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      421 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      481 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      541 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      601 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      661 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      721 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      781 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      841 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
      901 gggagaccca agctggctag cgtttaaacg ggccctctag actcgagatg agcgagctga
      961 ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca
     1021 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg
     1081 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg
     1141 gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc
     1201 ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta
     1261 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga
     1321 acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg
     1381 agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg
     1441 tgggcgggag ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta
     1501 agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg
     1561 ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc
     1621 ctagcaaact ggggcacgat ggaagcggag ctactaactt cagcctgctg aagcaggctg
     1681 gagacgtgga ggagaaccct ggacctatgg attccacaaa ctacatctcc atactgtgcg
     1741 agtatgctca gagacaacgt cagatctctg acatcaagtt tgaggaggtt ggaacagaag
     1801 gaccggatca cctgaaaacc ttcactctga gggtggtgat taaaggccat gcctatccta
     1861 acggggttgg aaagaacaag aaagaggcca aacagaatgc tgctaaacat gctctggctg
     1921 gcatgcctcc tgtccgagca gttggccgtc tgggatggag aaagcagagg tctggaatga
     1981 tgtcagaagc cagagttttc ctccccagtt caacacgcag ttcaacttag agaacaaggt
     2041 gatagagtcg atgctgtgtg ccaacccaga ggatagacca gatgccaggc agctgaagat
     2101 aaaactgatt gagtgttctt gtgttctgac cagagatcaa aacttccatc aagacaacag
     2161 aacagtctaa aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
     2221 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2281 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2341 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2401 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2461 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2521 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2581 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2641 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2701 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2761 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2821 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2881 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2941 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3061 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3121 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3181 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3241 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     3301 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     3361 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     3421 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     3481 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     3541 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     3601 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     3661 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     3721 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3781 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3841 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3961 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4021 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4081 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4141 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4201 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4321 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4381 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4441 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4501 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4561 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4621 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4681 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4741 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4801 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4861 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4921 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4981 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5041 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5101 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6061 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccc
//
