LOCUS       sequence-423960-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Blue Fluorescent Protein - P2A - full length 
            protein kinase R from chinook salmon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7891)
  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 7891)
  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-FL
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(116..184,185..976)
                     /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     complement(334..353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1147..1735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1636..1655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1889..1906
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2023..2044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2059..2064,2065..2082,2083..2089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2097..2113
                     /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)."
     primer_bind     2102..2124
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2121..2137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2121..2137
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2186..2307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2197..2216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2251..2270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2437..2811)
                     /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"
     promoter        complement(2830..2877)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2925..3254)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2939..3074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2993..3012)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3239..3259
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3268..3696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3378..3399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3590..3609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3742..3966
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3955..3972
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             5354..5365
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(6186..6242)
                     /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(6989..7008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6990..7008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7053..7256)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7082..7102)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7257..7636
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7808..7827
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
       61 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      301 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      421 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
      661 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      721 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      961 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1921 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1981 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2041 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2101 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     2161 agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
     2221 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2281 accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc
     2341 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     2401 gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag tcctgctcct cggccacgaa
     2461 gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc
     2521 gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca
     2581 ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac
     2641 cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa
     2701 gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc
     2761 ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtttagtt
     2821 cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat tgtcaacacg
     2881 tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca aaagcctagg
     2941 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc ggcctcggcc
     3001 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     3061 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     3121 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     3181 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     3241 cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa atcagctcat
     3301 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3361 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3421 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3481 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3541 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3601 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3661 cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc
     3721 tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt
     3781 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     3841 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     3901 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     3961 gcacagtcga ggctgatcag cggtttaaac ttaagctttt agactgttct gttgtcttga
     4021 tggaagtttt gatctctggt cagaacacaa gaacactcaa tcagttttat cttcagctgc
     4081 ctggcatctg gtctatcctc tgggttggca cacagcatcg actctatcac cttgttctct
     4141 aagttgaact gcgtgttgaa ctggggagga aaactctggc ttctgacatc attccagacc
     4201 tctgctttct ccatcccaga caggttccag agcagttcga agtagatcag acccaaagca
     4261 aagatgtcca cctttctgtc ataggaggtc tggttcctct gttcgggggc catgtaagac
     4321 ttggttcctg tctttttcgt tctctccagc aggttctcat cgttgtcgtt atcttctgcc
     4381 gtcaccagac caaagtcccc aatctttacc tctcccttcc cctctccgtc actcataccg
     4441 aacatgatgt tagcaggctt taggtctctg tgtaggagtt tcttggagtg gatgtattct
     4501 accccgttta ctatctgctg ggtgatgtgt agactctcct ccctgcgttt gggtttccga
     4561 tgagcgttcc tctcgtcgat ccacactttt agagtcctct tgtcacatag ttccatctgg
     4621 atgtacagga actcagagga cgaactgctg cctgaatctg aagtggtgtc gctctcagag
     4681 ttggtgtcgc atcggtacgc ggtgtcctcc agccaggcgg tgtagtagcg tactatgttg
     4741 gggtgctgaa gatctgctaa ggcaccaacc tcccgcttag ctttcccttt actcagtact
     4801 atcttcacgg caaaatactt ctgctccagt tcccgtcgcg ccttataaac gttgccaaag
     4861 cctcctttgc caatcttctc tattgaatca aactctgata aaaacctaga tttaattggt
     4921 tggttggatg tccttccact tgacaaggtt cctaaattgt tgccttttaa atcgggacct
     4981 tcctgtttgg tcttgaccgg tgacatcagg aagttaggtg ctagtcttat tttaggcttc
     5041 acatttacag ggctttcaac cttgggggag ctgacgttaa cggagtctgc gaatatgatg
     5101 gactcactgg tacttcctgg aatactcaag gatgatggtg ttgcatcctg ggattcccat
     5161 gtactggagg gggttagaga ggatagtcct ccatcttctg aaactgttga tctgcaagac
     5221 acctggctgt tccagtcaga ctgttcttga agagcagacc atgctaactg agcagcttgt
     5281 tgtttggcct gtttggctgt cttacccaaa ccatttgggt attctctctg gtcaatcaat
     5341 actttataaa caaattgagg gtcgtgtgac ggtccgctct tctccaccag tttaaagtca
     5401 ggaaaccgtt tggttttctg acagtagtgg ttgagtatac ctatgaagtt ggtctcctct
     5461 gttgtaatga agctgtatcg tgctggaaca ttctgttctg ggctcatgtt catgtgttca
     5521 accttctcac tgaggaccac aacctttgga ttcaattcct ccttgtgttt tccagtggtg
     5581 ccattgcagt tttcatctgc agtccctgta gtctgactac ggccacataa ctccagatag
     5641 atctgcattg ctgcttcctc cttagcctcc ttcttcgtgt tcccaaagcc ctctgggtat
     5701 tctttctcac ctaccacata cctacagcat tgtgaggtgt tgttgggtcc aacacgcgtt
     5761 tcctccaaag ccttcagaga cagcttgttc ttctggctgt gttcattcag ccagcacacg
     5821 tagttgggct gggagataac tgctcggaca ggaggggctg gagactgagc cggtgacgga
     5881 ggacatgaaa ccgatgcatc cttctgctct gtcgtctccg ccactccagc cagagcatgt
     5941 tcatccttct gctctgtcgt ctccatcatg ccagccagag catgtttagc agcattctgt
     6001 ttggcctctt tcttgttctt tccaaccccg ttaggatagg catggccttt aatcaccacc
     6061 ctcagagtga aggttttcag gtgatccggt ccttctgttc caacctcctc aaacttgatg
     6121 tcagagatct gacgttgtct ctgagcatac tcgcacagta tggagatgta gtttgtggaa
     6181 tccataggtc cagggttctc ctccacgtct ccagcctgct tcagcaggct gaagttagta
     6241 gctccgcttc catcgtgccc cagtttgcta gggaggtcgc agtatctggc cactgccacc
     6301 tcgtgctgct cgacgtaggt ctcgttgttg gcctccttga ttctttccag tctgtagtcc
     6361 acatagtaga cgccaggcat cttgaggttc ttagcgggtt tcttggatct atatgtggtc
     6421 ttgatgtttg cgatcagatg gctcccgccc acgagcttca gggccatgtc gtttctgcct
     6481 tccaggccgc cgtcagcggg gtacagcgtc tcggtgaagg cctcccagcc gagtgttttc
     6541 ttctgcatca cagggccgtt ggatgtgaag ttcacccctc tgatcttgac gttgtagatg
     6601 aggcagccgt cctggaggct ggtgtcctgg gtagcggtca gcacgccccc gtcttcgtat
     6661 gtggtgactc tctcccatgt gaagccctca gggaaggact gcttgaagaa gtcggggatg
     6721 ccctgggtgt ggttgatgaa ggtcttgctg ccgtagagga agctagtagc caggatgtcg
     6781 aaggcgaagg ggagagggcc gccctcgacc accttgattc tcatggtctg ggtgccctcg
     6841 tagggcttgc cttcgccctc ggatgtgcac ttgaagtgat ggttgtccac ggtgccctcc
     6901 atgtacagct tcatgtgcat gttctcctta atcagctcgc tcatctcgag tctagagggc
     6961 ccgtttaaac gctagccagc ttgggtctcc ctatagtgag tcgtattaat ttcgataagc
     7021 cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac ctcccaccgt
     7081 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     7141 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     7201 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     7261 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     7321 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     7381 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     7441 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     7501 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     7561 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     7621 gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc
     7681 ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa
     7741 attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt
     7801 aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc
     7861 ccgatccgtc gacgtcaggt ggcacttttc g
//
