LOCUS       sequence-228101-        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 wt
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2216..2225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2222..2224,2225..2227,2228..2941)
                     /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(2222..2224,2225..2227,2228..2938)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2267..2288)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2528..2547)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2875..2896
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3064..3185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3155..3174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3192..3647)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3329..3350)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3541..3560
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3780..4137
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3780..3971
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3803..3823)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3988..4123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4172..4966
                     /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             4172..4966
                     /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(4226..4245)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4836..4855
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5154..5173)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5198..5245
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5574..6162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6063..6082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctgtc gacatgtcgt gcgtcaagtt atggcccagc ggtgcccccg cccccttggt
      661 gtccatcgag gaactggaga accaggagct cgtcggcaaa ggcgggttcg gcacagtgtt
      721 ccgggcgcaa cataggaagt ggggctacga tgtggcggtc aagatcgtaa actcgaaggc
      781 gatatccagg gaggtcaagg ccatggcaag tctggataac gaattcgtgc tgcgcctaga
      841 aggggttatc gagaaggtga actgggacca agatcccaag ccggctctgg tgactaaatt
      901 catggagaac ggctccttgt cggggctgct gcagtcccag tgccctcggc cctggccgct
      961 cctttgccgc ctgctgaaag aagtggtgct tgggatgttt tacctgcacg accagaaccc
     1021 ggtgctcctg caccgggacc tcaagccgtc caacgtcctg ctggacccag agctgcacgt
     1081 caagctggca gattttggcc tgtccacatt tcagggaggc tcacagtcag ggacagggtc
     1141 cggggagcca gggggcaccc tgggctactt ggccccagaa ctgtttgtta acgtaaaccg
     1201 gaaggcctcc acagccagtg acgtctacag cttcgggatc ctaatgtggg cagtgcttgc
     1261 tggaagagaa gttgagttgc caaccgaacc atcactcgtg tacgaagcag tgtgcaacag
     1321 gcagaaccgg ccttcattgg ctgagctgcc ccaagccggg cctgagactc ccggcttaga
     1381 aggactgaag gagctaatgc agctctgctg gagcagtgag cccaaggaca gaccctcctt
     1441 ccaggaatgc ctaccaaaaa ctgatgaagt cttccagatg gtggagaaca atatgaatgc
     1501 tgctgtctcc acggtaaagg atttcctgtc tcagctcagg agcagcaata ggagattttc
     1561 tatcccagag tcaggccaag gagggacaga aatggatggc tttaggagaa ccatagaaaa
     1621 ccagcactct cgtaatgatg tcatggtttc tgagtggcta aacaaactga atctagagga
     1681 gcctcccagc tctgttccta aaaaatgccc gagccttacc aagaggagca gggcacaaga
     1741 ggagcaggtt ccacaagcct ggacagcagg cacatcttca gattcgatgg cccaacctcc
     1801 ccagactcca gagacctcaa ctttcagaaa ccagatgccc agccctacct caactggaac
     1861 accaagtcct ggaccccgag ggaatcaggg ggctgagaga caaggcatga actggtcctg
     1921 caggaccccg gagccaaatc cagtaacagg gcgaccgctc gttaacatat acaactgctc
     1981 tggggtgcaa gttggagaca acaactactt gactatgcaa cagacaactg ccttgcccac
     2041 atggggcttg gcaccttcgg gcaaggggag gggcttgcag caccccccac cagtaggttc
     2101 gcaagaaggc cctaaagatc ctgaagcctg gagcaggcca cagggttggt ataatcatag
     2161 cgggaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac
     2221 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2281 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2341 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2401 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2461 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2521 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2581 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2641 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2701 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2761 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2821 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2881 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2941 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     3001 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3061 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3121 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3181 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3241 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3301 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3361 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3421 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3481 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3541 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3601 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3781 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3841 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3961 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4021 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4081 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4141 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4201 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4261 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4321 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4381 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4441 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4501 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4561 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4621 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4681 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4741 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4801 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4861 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4921 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4981 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5041 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5101 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5161 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5221 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5281 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5341 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5401 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6241 cctgattctg tggataaccg tattaccgcc atgcattagt tat
//
