LOCUS       sequence-9520-ex        7770 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 7770)
  AUTHORS   Li S, Wang L, Dorf ME
  TITLE     PKC phosphorylation of TRAF2 mediates IKKalpha/beta recruitment and 
            K63-linked polyubiquitination.
  JOURNAL   Mol Cell. 2009 Jan 16. 33(1):30-42.
  PUBMED    19150425
REFERENCE   2  (bases 1 to 7770)
  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: "Mol Cell. 
            2009 Jan 16. 33(1):30-42."
COMMENT     Sequence Label: hTRAF2T117ApLPCX-HA-Flag/P875
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1566..2165
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(1566..1585)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2062..2082
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2340..2643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2644..2847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2798..2818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2907..2933
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4437..4460
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             4509..5102
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5205..5227)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5355..5388
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5448..5481
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5501..5520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5601..5618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5772..6360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5852..5871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6531..7322,7323..7391))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7154..7173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7392..7496)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7564..7582
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattgatccc ccgggctgca ggtcggccgc
     1501 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac
     1561 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     1621 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     1681 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     1741 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     1801 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     1861 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     1921 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     1981 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     2041 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2101 ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg
     2161 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct
     2221 ccgaccgaag ccgacccggg cggccccgcc gaccccgcac ccgcccccga ggcccaccga
     2281 ctctagtaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     2341 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2401 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2461 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     2521 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     2581 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     2641 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     2701 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     2761 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     2821 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     2881 cggactcaga tctcgacgcc accatgtacc catacgatgt tccagattac gctcttgctg
     2941 cagctagcgt gaccccccct ggctccctgg agttgctaca gcccggcttc tccaagaccc
     3001 tcctggggac caagctggaa gccaagtacc tgtgctccgc ctgcagaaac gtcctccgca
     3061 ggcccttcca ggcgcagtgt ggccaccggt actgctcctt ctgcctggcc agcatcctca
     3121 gctctgggcc tcagaactgt gctgcctgtg ttcacgaggg catatatgaa gaaggcattt
     3181 ctattttaga aagcagttcg gccttcccag ataatgctgc ccgcagggag gtggagagcc
     3241 tgccggccgt ctgtcccagt gatggatgca cctggaaggg ggccctgaaa gaatacgaga
     3301 gctgccacga aggccgctgc ccgctcatgc tgaccgaatg tcccgcgtgc aaaggcctgg
     3361 tccgccttgg tgaaaaggag cgccacctgg agcacgagtg cccggagaga agcctgagct
     3421 gccggcattg ccgggcaccc tgctgcggag cagacgtgaa ggcgcaccac gaggtctgcc
     3481 ccaagttccc cttaacttgt gacggctgcg gcaagaagaa gatcccccgg gagaagtttc
     3541 aggaccacgt caagacttgt ggcaagtgtc gagtcccttg cagattccac gccatcggct
     3601 gcctcgagac ggtagagggt gagaaacagc aggagcacga ggtgcagtgg ctgcgggagc
     3661 acctggccat gctactgagc tcggtgctgg aggcaaagcc cctcttggga gaccagagcc
     3721 acgcggggtc agagctcctg cagaggtgcg agagcctgga gaagaagacg gccacttttg
     3781 agaacattgt ctgcgtcctg aaccgggagg tggagagggt ggccatgact gccgaggcct
     3841 gcagccggca gcaccggctg gaccaagaca agattgaagc cctgagtagc aaggtgcagc
     3901 agctggagag gagcattggc ctcaaggacc tggcgatggc tgacttggag cagaaggtct
     3961 tggagatgga ggcatccacc tacgatgggg tcttcatctg gaagatctca gacttcgcca
     4021 ggaagcgcca ggaagctgtg gctggccgca tacccgccat cttctcccca gccttctaca
     4081 ccagcaggta cggctacaag atgtgtctgc gtatctacct gaacggcgac ggcaccgggc
     4141 gaggaacaca cctgtccctc ttctttgtgg tgatgaaggg cccgaatgac gccctgctgc
     4201 ggtggccctt caaccagaag gtgaccttaa tgctgctcga ccagaataac cgggagcacg
     4261 tgattgacgc cttcaggccc gacgtgactt catcctcttt tcagaggcca gtcaacgaca
     4321 tgaacatcgc aagcggctgc cccctcttct gccccgtctc caagatggag gcaaagaatt
     4381 cctacgtgcg ggacgatgcc atcttcatca aggccattgt ggacctgaca gggctcgatt
     4441 acaaggatga cgacgataag tagatcgata aaataaaaga ttttatttag tctccagaaa
     4501 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     4561 tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca
     4621 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     4681 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     4741 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag
     4801 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     4861 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     4921 cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga
     4981 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     5041 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     5101 tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg
     5161 aggtaagctg gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5221 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5281 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     5341 agcggagtgt actgataact tcgtataatg tatgctatac gaagttatta ggtctgaaga
     5401 ggagtttacg tccagccaag ctagggccgc gatccggaac ccttaatata acttcgtata
     5461 atgtatgcta tacgaagtta tcagtactgg cttaactatg cggcatcaga gcagattgta
     5521 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5581 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5641 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6181 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6841 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7141 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7381 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7441 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7501 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7561 aaaaataggc gtatcacgag gccctttcgt cttcaagaat tgctagcaat tgctagcaat
     7621 tgctagcaat tcataccaga tcaccgaaaa ctgtcctcca aatgtgtccc cctcacactc
     7681 ccaaattcgc gggcttctgc ctcttagacc actctaccct attccccaca ctcaccggag
     7741 ccaaagccgc ggcccttccg tttctttgct
//
