LOCUS       sequence-337647-        6463 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 6463)
  AUTHORS   Zhu G, Wu CJ, Zhao Y, Ashwell JD
  TITLE     Optineurin negatively regulates TNFalpha- induced NF-kappaB 
            activation by competing with NEMO for ubiquitinated RIP.
  JOURNAL   Curr Biol. 2007 Aug 21. 17(16):1438-43.
  PUBMED    17702576
REFERENCE   2  (bases 1 to 6463)
  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: "Curr Biol. 
            2007 Aug 21. 17(16):1438-43."
COMMENT     Sequence Label: pDEST17-hOPTN
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             499..510
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1932..1956)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2009..2027)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2023..2070
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2436..2441,2442..2458,2459..2464))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2491..2509)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2577..2681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2682..2750,2751..3542)
                     /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     complement(2900..2919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3713..4301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4202..4221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4455..4472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4487..4626
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4552..4571)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4712..4734
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4728..4919)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6381..6400
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggctccgcgg ccgccccctt
      181 cacctcccat caacctctca gctgcctcac tgaaaaggag gacagcccca gtgaaagcac
      241 aggaaatgga cccccccacc tggcccaccc aaacctggac acgtttaccc cggaggagct
      301 gctgcagcag atgaaagagc tcctgaccga gaaccaccag ctgaaagaag ccatgaagct
      361 aaataatcaa gccatgaaag ggagatttga ggagctttcg gcctggacag agaaacagaa
      421 ggaagaacgc cagttttttg agatacagag caaagaagca aaagagcgtc taatggcctt
      481 gagtcatgag aatgagaaat tgaaggaaga gcttggaaaa ctaaaaggga aatcagaaag
      541 gtcatctgag gaccccactg atgactccag gcttcccagg gccgaagcgg agcaggaaaa
      601 ggaccagctc aggacccagg tggtgaggct acaagcagag aaggcagacc tgttgggcat
      661 cgtgtctgaa ctgcagctca agctgaactc cagcggctcc tcagaagatt cctttgttga
      721 aattaggatg gctgaaggag aagcagaagg gtcagtaaaa gaaatcaagc atagtcctgg
      781 gcccacgaga acagtctcca ctggcacggc attgtctaaa tataggagca gatctgcaga
      841 tggggccaag aattacttcg aacatgagga gttaactgtg agccagctcc tgctgtgcct
      901 aagggaaggg aatcagaagg tggagagact tgaagttgca ctcaaggagg ccaaagaaag
      961 agtttcagat tttgaaaaga aaacaagtaa tcgttctgag attgaaaccc agacagaggg
     1021 gagcacagag aaagagaatg atgaagagaa aggcccggag actgttggaa gcgaagtgga
     1081 agcactgaac ctccaggtga catctctgtt taaggagctt caagaggctc atacaaaact
     1141 cagcgaagct gagctaatga agaagagact tcaagaaaag tgtcaggccc ttgaaaggaa
     1201 aaattctgca attccatcag agttgaatga aaagcaagag cttgtttata ctaacaaaaa
     1261 gttagagcta caagtggaaa gcatgctatc agaaatcaaa atggaacagg ctaaaacaga
     1321 ggatgaaaag tccaaattaa ctgtgctaca gatgacacac aacaagcttc ttcaagaaca
     1381 taataatgca ttgaaaacaa ttgaggaact aacaagaaaa gagtcagaaa aagtggacag
     1441 ggcagtgctg aaggaactga gtgaaaaact ggaactggca gagaaggctc tggcttccaa
     1501 acagctgcaa atggatgaaa tgaagcaaac cattgccaag caggaagagg acctggaaac
     1561 catgaccatc ctcagggctc agatggaagt ttactgttct gattttcatg ctgaaagagc
     1621 agcgagagag aaaattcatg aggaaaagga gcaactggca ttgcagctgg cagttctgct
     1681 gaaagagaat gatgctttcg aagacggagg caggcagtcc ttgatggaga tgcagagtcg
     1741 tcatggggcg agaacaagtg actctgacca gcaggcttac cttgttcaaa gaggagctga
     1801 ggacagggac tggcggcaac agcggaatat tccgattcat tcctgcccca agtgtggaga
     1861 ggttctgcct gacatagaca cgttacagat tcacgtgatg gattgcatca tttaaaaggg
     1921 tgggcgcgcc gacccagctt tcttgtacaa agtggttgat tcgaggctgc taacaaagcc
     1981 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2041 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatcca
     2101 caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag
     2161 caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa
     2221 ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg
     2281 gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc
     2341 cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg
     2401 actgcgttag caatttaact gtgataaact accgcattaa agcttatcga tgataagctg
     2461 tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2521 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2581 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2641 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2701 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2761 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2821 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2881 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2941 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3001 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3061 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3121 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3181 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     3241 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3301 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3361 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3421 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3481 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3541 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3601 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3661 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3781 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3841 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3901 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3961 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4021 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4081 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4141 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4201 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4261 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4321 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4381 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4441 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4501 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     4561 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     4621 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4681 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4741 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4801 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4861 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4921 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     4981 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     5041 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     5101 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     5161 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     5221 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     5281 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     5341 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     5401 ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     5461 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     5521 gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
     5581 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg
     5641 ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa
     5701 tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt
     5761 catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga
     5821 gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc
     5881 ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     5941 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     6001 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     6061 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     6121 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     6181 gatcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     6241 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     6301 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     6361 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     6421 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcg
//
