LOCUS       sequence-343216-        8972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8972)
  TITLE     Sun Lab Overexpression Plasmids
REFERENCE   2  (bases 1 to 8972)
  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: pCLXSN(GFP)-mTRAF2
FEATURES             Location/Qualifiers
     source          1..8972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(62..64,65..67,68..781)
                     /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(62..64,65..67,68..778)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(107..128)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(368..387)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     715..736
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             887..1480
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          1825..1890
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2020..2040
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2312..2446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2349..2368)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2403..2422
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2904..3038
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3103..3120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3274..3862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3354..3373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4033..4824,4825..4893))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4656..4675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4894..4998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5066..5084
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     5244..5263
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5408..5787
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5788..5991
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5942..5962
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5992..6167
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    6230..6429
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6630..7046
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6976..6998
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7014..7030
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             7084..7110
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(8641..8661)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8646..8972,1..3)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8826..8961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaagcttg ggctgcaggt cgaggcggat ctgatcaaga gacaggatga gcatcgatac
       61 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      121 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      181 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      241 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      301 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      361 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      421 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      481 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      541 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      601 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      661 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
      721 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
      781 agatgcatcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
      841 gttcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
      901 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
      961 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     1021 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     1081 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     1141 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     1201 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     1261 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     1321 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     1381 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     1441 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     1501 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc
     1561 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1621 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     1681 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat ctagctaggt
     1741 agctagagga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta
     1801 cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa
     1861 ctactgattc taattgtttg tgtattttag attccaacct atggaactga tgaatgggag
     1921 cagtggtgga atgcctttaa tgaggaaaac ctgttttgct cagaagaaat gccatctagt
     1981 gatgatgagg ctactgctga ctctcaacat tctactcctc caaaaaagaa gagaaaggta
     2041 gaagacccca aggactttcc ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt
     2101 aatagaactc ttgcttgctt tgctatttac accacaaagg aaaaagctgc actgctatac
     2161 aagaaaatta tggaaaaata tttgatgtat agtgccttga ctagagatca taatcagcca
     2221 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2281 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2341 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2401 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcagct tcagaagatg
     2461 gcggagggcc tccaacacag taattttcct cccgactctt aaaatagaaa atgtcaagtc
     2521 agttaagcag gaagtgacta actgaccgca gctggccgtg cgacatcctc ttttaattag
     2581 ttgctaggca actgccctcc agagggcagt gtggttttgc aagaggaagc aaaaagcctc
     2641 tccacccagg cctagaatgt ttccacccaa tcattactat gacaacagct gtttttttta
     2701 gtattaagca gaggccgggg gcccctggcc tccgcttact ctggagaaaa agaagagagg
     2761 cattgtagag gcttccagag gcaacttgtc aaaacaggac tgcttctatt tctgtcacac
     2821 tgtctggccc tgtcacaagg tccagcacct ccataccccc tttaataagc agtttgggaa
     2881 cgggtgcggg tcttactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2941 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3001 agctattcca gaagtagtga ggaggctttt ttggaggctg cattaatgaa tcggccaacg
     3061 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3181 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3241 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4141 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4321 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     4381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4621 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4861 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4981 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5041 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5101 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5161 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5221 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5281 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     5341 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5401 gcgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     5461 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5521 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5581 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     5641 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     5701 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5761 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     5821 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     5881 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     5941 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcaataaaag
     6001 agcccacaac ccctcactcg gcgcgccagt cttccgatag actgcgtcgc ccgggtaccc
     6061 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggtctcgct gttccttggg
     6121 agggtctcct ctgagtgatt gactacccac gacgggggtc tttcatttgg gggctcgtcc
     6181 gggatttgga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag
     6241 caacttatct gtgtctgtcc gattgtctag tgtctatgtt tgatgttatg cgcctgcgtc
     6301 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     6361 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     6421 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     6481 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     6541 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     6601 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     6661 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     6721 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     6781 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     6841 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     6901 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     6961 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     7021 cagccctcac tccttctcta ggcgccggaa ttcagatctg gtaccacgcg taccatggat
     7081 atctatccat atgacgttcc agattatgct gctcagatcg atatggctgc agccagtgtg
     7141 acttcccctg gctccctaga actgctacag cctggcttct ccaagaccct cctggggacc
     7201 aggttagaag ccaagtacct ctgttcagcc tgcaaaaaca tcctgcggag gcctttccag
     7261 gcccagtgtg ggcaccgcta ctgctccttc tgcctgacca gcatcctcag ctctgggccc
     7321 cagaactgtg ctgcctgtgt ctatgaaggc ctgtatgaag aaggcatttc tattttagag
     7381 agtagttcgg cctttccaga taacgctgcc cgcagagagg tggagagcct gccagctgtc
     7441 tgtcccaatg atggatgcac ttggaagggg accttgaaag aatacgagag ctgccacgaa
     7501 ggactttgcc cattcctgct gacggagtgt cctgcatgta aaggcctggt ccgcctcagc
     7561 gagaaggagc accacactga gcaggaatgc cccaaaagga gcctgagctg ccagcactgc
     7621 agagcaccct gtagccacgt ggacctggag gtacactatg aggtctgccc caagtttccc
     7681 ttaacctgtg atggctgtgg caagaagaag atccctcggg agacgtttca ggaccatgtt
     7741 agagcatgca gcaaatgccg ggttctctgc agattccaca ccgttggctg ttcagagatg
     7801 gtggagactg agaacctgca ggatcatgag ctgcagcggc tacgggaaca cctagcccta
     7861 ctgctgagct cattcttgga ggcccaagcc tctccaggaa ccttgaacca ggtggggcca
     7921 gagctactcc agcggtgcca gattttggag cagaagatag caacctttga gaacattgtc
     7981 tgcgtcttga accgtgaagt agagagggta gcagtgactg cagaggcttg tagccggcag
     8041 caccggctag accaggacaa gattgaggcc ctgagtaaca aggtgcaaca gctggagagg
     8101 agcatcggcc tcaaggacct ggccatggct gacctggagc agaaggtctc cgagttggaa
     8161 gtatccacct atgatggggt cttcatctgg aagatctctg acttcaccag aaagcgtcag
     8221 gaagccgtag ctggccggac accagctatc ttctccccag ccttctacac aagcagatat
     8281 ggctacaaga tgtgtctacg agtctacttg aatggcgacg gcactgggcg gggaactcat
     8341 ctgtctctct tcttcgtggt gatgaaaggc cccaatgatg ctctgttgca gtggcctttt
     8401 aatcagaagg taacattgat gttgctggac cataacaacc gggagcatgt gatcgacgca
     8461 ttcaggcccg atgtaacctc gtcctccttc cagaggcctg tcagtgacat gaacatcgcc
     8521 agtggctgcc ccctcttctg ccctgtgtcc aagatggagg ccaagaattc ctatgtgcgg
     8581 gatgatgcga tcttcatcaa agctattgtg gacctaacag gactctaggg atccggctgt
     8641 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     8701 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     8761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     8821 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     8881 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     8941 gaggaggctt ttttggaggc ctaggctttt gc
//
