LOCUS       sequence-423973-        6826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Blue Fluorescent Protein - P2A - medium length 
            protein kinase R from chinook salmon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6826)
  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 6826)
  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-ML
FEATURES             Location/Qualifiers
     source          1..6826
                     /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(2855..2872)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2861..3085
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3131..3559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3218..3237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3428..3449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3568..3588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3573..3902
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3753..3888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3815..3834
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3950..3997
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4016..4390
                     /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    4520..4641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4557..4576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4611..4630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4690..4706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4690..4706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4703..4725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4714..4730
                     /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(4738..4744,4745..4762,4763..4768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4783..4804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4921..4938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5092..5680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5172..5191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5851..6642,6643..6711))
                     /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     6474..6493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6712..6816)
                     /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 gcatgatgga gacgacagag cagaaggatg aacatgctct ggctggagtg gcggagacga
     1981 cagagcagaa ggatgcatcg gtttcatgtc ctccgtcacc ggctcagtct ccagcccctc
     2041 ctgtccgagc agttatctcc cagcccaact acgtgtgctg gctgaatgaa cacagccaga
     2101 agaacaagct gtctctgaag gctttggagg aaacgcgtgt tggacccaac aacacctcac
     2161 aatgctgtag ttcaggcagc agttcgtcct ctgagttcct gtacatccag atggaactat
     2221 gtgacaagag gactctaaaa gtgtggatcg acgagaggaa cgctcatcgg aaacccaaac
     2281 gcagggagga gagtctacac atcacccagc agatagtaaa cggggtagaa tacatccact
     2341 ccaagaaact cctacacaga gacctaaagc ctgctaacat catgttcggt atgagtgacg
     2401 gagaggggaa gggagaggta aagattgggg actttggtct ggtgacggca gaagataacg
     2461 acaacgatga gaacctgctg gagagaacga aaaagacagg aaccaagtct tacatggccc
     2521 ccgaacagag gaaccagacc tcctatgaca gaaaggtgga catctttgct ttgggtctga
     2581 tctacttcga actgctctgg aacctgtctg ggatggagaa agcagaggtc tggaatgatg
     2641 tcagaagcca gagttttcct ccccagttca acacgcagtt caacttagag aacaaggtga
     2701 tagagtcgat gctgtgtgcc aacccagagg atagaccaga tgccaggcag ctgaagataa
     2761 aactgattga gtgttcttgt gttctgacca gagatcaaaa cttccatcaa gacaacagaa
     2821 cagtctaaaa gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     2881 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2941 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3001 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3061 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3121 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3181 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3241 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3301 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3361 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3421 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3481 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3541 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3781 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3841 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3901 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     3961 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     4021 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     4081 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     4141 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     4201 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     4261 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     4321 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     4381 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4501 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4621 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5521 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5761 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5821 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5881 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5941 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6001 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6061 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6121 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6181 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6241 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6301 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6361 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6421 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6481 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6541 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6601 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6661 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6721 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6781 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccc
//
