LOCUS       sequence-298554-        6181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6181)
  AUTHORS   Cho YS, Challa S, Moquin D, Genga R, Ray TD, Guildford M, Chan FK
  TITLE     Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates 
            programmed necrosis and virus-induced inflammation.
  JOURNAL   Cell. 2009 Jun 12;137(6):1112-23.
  PUBMED    19524513
REFERENCE   2  (bases 1 to 6181)
  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: "Cell."; 
            date: "2009-06-12"; volume: "137(6)"; pages: "1112-23"
COMMENT     Sequence Label: mRIP3 GFP RHIM mut
FEATURES             Location/Qualifiers
     source          1..6181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2824..2833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2830..2832,2833..2835,2836..3549)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2830..2832,2833..2835,2836..3546)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2875..2896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3136..3155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3483..3504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3672..3793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3709..3728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3763..3782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3800..4255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3937..3958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4149..4168
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4282..4386
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4388..4745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4388..4579
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4411..4431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4596..4731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4780..5574
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4780..5574
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4834..4853)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5444..5463
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5762..5781)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5806..5853
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctact
     1321 atgtcttctg tcaagttatg gcctactggt gcgtcagcgg ttcctctggt gagccgtgaa
     1381 gaactgaaga agctggagtt tgtgggcaaa ggagggttcg gagtcgtgtt ccgggcacac
     1441 cacagaacat ggaaccatga tgtagcagtc aagatcgtga actcgaagaa gatatcctgg
     1501 gaggtgaagg ctatggttaa tcttcgtaat gagaacgttc tgctcctgct gggggtcact
     1561 gaggacctcc agtgggactt cgtgtccggg caggctctgg tgacaagatt catggagaat
     1621 ggctccctcg cagggctgct gcaacccgag tgccctcggc cctggccact cctctgtcgc
     1681 ctgctgcagg aagtggtgct ggggatgtgc tacctacaca gcttggaccc tccgctcctg
     1741 caccgggacc tcaagccctc taacattctg ctggatccag agctccacgc caagctagca
     1801 gattttggcc tgtccacgtt tcagggaggg tcccagtcag ggtcaggatc aggatcagga
     1861 tccagggact ctgggggcac cctagcgtac ttggacccag agctgttatt taaagtcaac
     1921 ctgaaggctt ctaaagcgag tgatgtctac agctttggga tcctcgtgtg ggcagtgctg
     1981 gctggcagag aagctgagtt ggtagacaag acttcactaa tccgggaaac agtgtgtgac
     2041 aggcagagtc gtcctccact gacagagctg cctccaggta gccctgagac tcccggcttg
     2101 gaaaaactga aggagttaat gattcattgc tggggttccc agtccgaaaa caggccatcc
     2161 ttccaggact gcgaaccaaa aaccaatgaa gtttacaatc tggtaaagga caaggtagat
     2221 gctgctgtct ccgaggtaaa gcattatctg tctcagcaca gaagcagtgg cagaaacttg
     2281 tctgccagag agccaagcca aagaggcaca gaaatggatt gcccgaggga aaccatggtt
     2341 tctaaaatgc tggaccgcct gcatttggag gaaccctccg gaccagttcc tggaaaatgt
     2401 cctgagaggc aagcacagga cacatcagtt gggcctgcca caccagcaag gacatcttct
     2461 gaccccgtgg ctggcactcc tcagattcca catactttac ccttcagagg cacaacacct
     2521 gggccagtct ttactgagac tcccggtcct cacccccaaa ggaatcaggg agatggaaga
     2581 cacggcactc cttggtatcc ctggacccca ccgaatccaa tgacagggcc accggctctc
     2641 gtcttcaaca actgttctga agtggcggct gcggcctaca actccttggt agcaccacca
     2701 agaactactg cctcaagttc ggccaagtat gaccaagcac agttcggcag gggtaggggc
     2761 tggcagccct tccacaagcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     2821 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2881 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2941 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3001 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3061 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3121 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3181 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3241 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3301 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3361 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3421 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3481 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3541 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     3601 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3661 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3721 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3781 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3841 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3901 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3961 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4021 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4081 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4141 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4201 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4381 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4441 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4501 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4561 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4621 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4681 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4741 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4801 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4861 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4921 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4981 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5041 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5101 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5161 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5221 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5281 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5341 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5401 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5461 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5521 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5581 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5641 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5701 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5761 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5821 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5881 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5941 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6001 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6181 c
//
