LOCUS       sequence-430350-        6917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 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 6917)
  TITLE     Collet Lab Plasmids 2024
REFERENCE   2  (bases 1 to 6917)
  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"
COMMENT     Sequence Label: pcDNA3.1-Pur-BFP-P2A-ppViperin
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    130..251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(167..186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     221..240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(300..316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(300..316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(313..335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    324..340
                     /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(348..354,355..372,373..378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    393..414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(531..548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(702..1290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(782..801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1461..2252,2253..2321))
                     /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     2084..2103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2322..2426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2501..2520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2692..3071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3072..3275
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3226..3246
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3320..3339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3320..3338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4086..4142
                     /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(5207..5224)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5213..5437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5483..5911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5570..5589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5780..5801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5920..5940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5925..6254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6105..6240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6167..6186
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6310..6909
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
       61 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
      121 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
      181 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
      241 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
      301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1141 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     2461 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     2521 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     2581 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     2641 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     2701 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     2761 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2821 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2881 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2941 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     3001 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     3061 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     3121 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     3181 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     3241 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     3301 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
     3361 taaacgggcc ctctagactc gagatgagcg agctgattaa ggagaacatg cacatgaagc
     3421 tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca
     3481 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
     3541 tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca
     3601 cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag
     3661 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
     3721 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga
     3781 tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg
     3841 gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa
     3901 acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct
     3961 actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc
     4021 agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg cacgatggaa
     4081 gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac
     4141 ctatgttgat gccattgtgt ttcaaggacg tccacagctt cttttcagcc ctgttgagat
     4201 ggattttgat gatggtatca ggcacactgg tgtctcttgg gatgatcagt cgtccaaaga
     4261 ttcgcaccag agagcagaaa gaggcatcca gcgcacaggt gaccactcca agcagtgtga
     4321 actaccattt tacccggcag tgcaattaca aatgtggctt ttgcttccac actgcgaaga
     4381 cctcgttcgt cttgcctatt gaagaggcaa agcgagggtt acgacttctg aaagaagcag
     4441 gaatggaaaa aatcaacttt tcaggtggag agccctttct tcatgagaga ggctcttttc
     4501 tgggagagtt ggtccgatac tgcaaacagg agttgctgct tccgagcgtc agcatcgtta
     4561 gtaatggcag tctgatcaaa gaatcctggt ttcagaaata cggtgactac ttggacattc
     4621 ttgcagtatc ttgcgatagt tttaacgaag acaccaataa agtcattggc cgaggtcagg
     4681 gcaagaagag ccatttagac aatctgcata aagtttgttc ctggtgccgg gactacaagg
     4741 tggctttcaa aatcaactcc gtgatcaaca cctacaatgt ggacgaagat atgacagagc
     4801 agatcactgc tctgaaccca gtgcgctgga aggtcttcca gtgtctgcta attgatggtg
     4861 aaaacgctgg ggagaacagc ctccgcgagg cagaaaaatt tgtcattagt gagcagcaat
     4921 tccaagactt cctggaccgc cataagagcg tcaagtgtct ggttccagag tctaatcaaa
     4981 agatgagaga ctcttacctg attcttgatg aatatatgcg cttcttggat tgccgagagg
     5041 ggaggaaaga tccgtcaaag tccgttctgg atgttggtgt ggaagaggcc atcaagttca
     5101 gtggttttga tgagaagatg ttcctcataa gaggggggaa atatgtgtgg agcaaggaag
     5161 atatgaaact ggagtggtga aagcttaagt ttaaaccgct gatcagcctc gactgtgcct
     5221 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5281 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     5341 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     5401 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     5461 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5881 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6001 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6061 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6181 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6241 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     6301 tgcctgagga tgaccgagta caagcctacc gtgcgcctgg ccactcgcga tgatgtgccc
     6361 cgcgccgtcc gcactctggc cgccgctttc gccgactacc ccgctacccg gcacaccgtg
     6421 gaccccgacc ggcacatcga gcgtgtgaca gagttgcagg agctgttcct gacccgcgtc
     6481 gggctggaca tcggcaaggt gtgggtagcc gacgacggcg cggccgtggc cgtgtggact
     6541 acccccgaga gcgttgaggc cggcgccgtg ttcgccgaga tcggcccccg aatggccgag
     6601 ctgagcggca gccgcctggc cgcccagcag caaatggagg gcctgcttgc cccccatcgt
     6661 cccaaggagc ctgcctggtt tctggccact gtaggagtga gccccgacca ccagggcaag
     6721 ggcttgggca gcgccgtcgt gttgcccggc gtagaggccg ccgaacgcgc cggtgtgccc
     6781 gcctttctcg aaacaagcgc accaagaaac cttccattct acgagcgcct gggcttcacc
     6841 gtgaccgccg atgtcgaggt gcccgaggga cctaggacct ggtgtatgac acgaaaacct
     6901 ggcgcctaag gcgcgcc
//
