LOCUS       sequence-298555-        6283 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 6283)
  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 6283)
  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: hRIP3 GFP D160N
FEATURES             Location/Qualifiers
     source          1..6283
                     /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      2926..2935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2932..2934,2935..2937,2938..3651)
                     /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(2932..2934,2935..2937,2938..3648)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2977..2998)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3238..3257)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3585..3606
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3774..3895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3811..3830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3865..3884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3902..4357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4039..4060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4251..4270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4384..4488
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4490..4847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4490..4681
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4513..4533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4698..4833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4882..5676
                     /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             4882..5676
                     /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(4936..4955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5546..5565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5864..5883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5908..5955
                     /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 cagatctgtc
     1321 gacatgtcgt gcgtcaagtt atggcccagc ggtgcccccg cccccttggt gtccatcgag
     1381 gaactggaga accaggagct cgtcggcaaa ggcgggttcg gcacagtgtt ccgggcgcaa
     1441 cataggaagt ggggctacga tgtggcggtc aagatcgtaa actcgaaggc gatatccagg
     1501 gaggtcaagg ccatggcaag tctggataac gaattcgtgc tgcgcctaga aggggttatc
     1561 gagaaggtga actgggacca agatcccaag ccggctctgg tgactaaatt catggagaac
     1621 ggctccttgt cggggctgct gcagtcccag tgccctcggc cctggccgct cctttgccgc
     1681 ctgctgaaag aagtggtgct tgggatgttt tacctgcacg accagaaccc ggtgctcctg
     1741 caccgggacc tcaagccgtc caacgtcctg ctggacccag agctgcacgt caagctggca
     1801 aattttggcc tgtccacatt tcagggaggc tcacagtcag ggacagggtc cggggagcca
     1861 gggggcaccc tgggctactt ggccccagaa ctgtttgtta acgtaaaccg gaaggcctcc
     1921 acagccagtg acgtctacag cttcgggatc ctaatgtggg cagtgcttgc tggaagagaa
     1981 gttgagttgc caaccgaacc atcactcgtg tacgaagcag tgtgcaacag gcagaaccgg
     2041 ccttcattgg ctgagctgcc ccaagccggg cctgagactc ccggcttaga aggactgaag
     2101 gagctaatgc agctctgctg gagcagtgag cccaaggaca gaccctcctt ccaggaatgc
     2161 ctaccaaaaa ctgatgaagt cttccagatg gtggagaaca atatgaatgc tgctgtctcc
     2221 acggtaaagg atttcctgtc tcagctcagg agcagcaata ggagattttc tatcccagag
     2281 tcaggccaag gagggacaga aatggatggc tttaggagaa ccatagaaaa ccagcactct
     2341 cgtaatgatg tcatggtttc tgagtggcta aacaaactga atctagagga gcctcccagc
     2401 tctgttccta aaaaatgccc gagccttacc aagaggagca gggcacaaga ggagcaggtt
     2461 ccacaagcct ggacagcagg cacatcttca gattcgatgg cccaacctcc ccagactcca
     2521 gagacctcaa ctttcagaaa ccagatgccc agccctacct caactggaac accaagtcct
     2581 ggaccccgag ggaatcaggg ggctgagaga caaggcatga actggtcctg caggaccccg
     2641 gagccaaatc cagtaacagg gcgaccgctc gttaacatat acaactgctc tggggtgcaa
     2701 gttggagaca acaactactt gactatgcaa cagacaactg ccttgcccac atggggcttg
     2761 gcaccttcgg gcaaggggag gggcttgcag caccccccac cagtaggttc gcaagaaggc
     2821 cctaaagatc ctgaagcctg gagcaggcca cagggttggt ataatcatag cgggaagctt
     2881 cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtgagc
     2941 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3001 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3061 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3121 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3181 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3241 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3301 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3361 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3421 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3481 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3541 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3601 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc
     3661 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3721 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3781 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3841 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3901 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3961 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4021 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4081 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4141 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4201 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4261 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4321 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     4381 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4441 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     4501 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4561 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4621 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4681 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4741 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4801 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4861 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4921 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4981 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5041 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5101 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5161 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5221 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5281 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5341 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5401 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5461 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5521 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5581 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5641 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5701 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccacagcc gccttctatg
     5761 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5821 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5881 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5941 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     6001 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     6061 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     6121 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
