LOCUS       40924_33787        6218 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pOPINJ, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R, 
            Rahman N, Stuart DI, Owens RJ.
  TITLE     A versatile ligation-independent cloning method suitable for 
            high-throughput expression screening applications
  JOURNAL   Nucleic Acids Res. 35 (6), E45 (2007)
  PUBMED    17317681
REFERENCE   2  (bases 1 to 6218)
  AUTHORS   Berrow NS.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JAN-2007) Oxford Protein Production Facility, Oxford 
            University, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K.
REFERENCE   3  (bases 1 to 6218)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6218)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2007"; volume: "35"; issue: "6"; pages: "E45"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JAN-2007) Oxford Protein Production Facility, Oxford University,
            Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K."
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     181..1008
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     enhancer        1140..1443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1449..1726
                     /label=chicken beta-actin promoter
     promoter        2150..2168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2169..2193
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        2209..2318
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     regulatory      2318..2326
                     /regulatory_class="ribosome_binding_site"
     CDS             2339..2356
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2357..3010
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     misc_feature    3033..3047
                     /label=5' in-fusion site (after KnpI digest)
                     /note="5' in-fusion site (after KnpI digest)"
     protein_bind    3093..3114
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3129..3159
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3167..3183
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             3203..3376
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    3385..3399
                     /label=3' in-fusion site (after HindIII digest)
                     /note="3' in-fusion site (after HindIII digest)"
     CDS             3406..3423
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    3578..3633
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      3721..3768
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3781..4486
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(4495..4528)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4696..5283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5299..6156)
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
       61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
      121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
      181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
      241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
      301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
      361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
      421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
      481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
      541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
      601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
      661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
      721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
      781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
      841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
      901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
      961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggccg
     1021 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctct
     1081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     1141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     1201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     1261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1441 atgcatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     1501 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     1561 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
     1621 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     1681 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     1741 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     1801 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctccttcgg gctgtaatta
     1861 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
     1921 ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc
     1981 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     2041 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     2101 ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt aatacgactc
     2161 actatagggg aattgtgagc ggataacaat tccccggagt taatccggga cctttaattc
     2221 aacccaacac aatatattat agttaaataa gaattattat caaatcattt gtatattaat
     2281 taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata ccatggcaca
     2341 tcaccatcac catcacatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     2401 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     2461 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     2521 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     2581 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     2641 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     2701 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     2761 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     2821 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     2881 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     2941 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     3001 tcctccaaaa tcggatctga gcagcggtct ggaagttctg tttcagggta ccgtcgaccc
     3061 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     3121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     3181 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     3241 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3301 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3361 cagcctgaat ggcgaatggc gccaagcttt ctagaccagt ttaaacatca ccatcaccat
     3421 cactaagtga ttaacctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     3481 atgccctggc tcacaaatac cactgagatc gatctttttc cctctgccaa aaattatggg
     3541 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     3601 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     3661 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgtaa
     3721 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaagcatgc
     3781 ggaggaaatt ctccttgaag tttccctggt gttcaaagta aaggagtttg caccagacgc
     3841 acctctgttc actggtccgg cgtattaaaa cacgatacat tgttattagt acatttatta
     3901 agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt ttaataattc
     3961 attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc
     4021 tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt ttcgattagt
     4081 ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat tttcgctcaa
     4141 cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat tcgtttgtgt
     4201 tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg tatttctttc
     4261 atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaatagag cttggacata
     4321 tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc
     4381 gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat taattgtgtc
     4441 ggctaacacg tccgcgatca aatttgtagt tgagcttttt ggaattgcga tcgcataact
     4501 tcgtatagca tacattatac gaagttataa gctcggaacg ctgcgctcgg tcgttcggct
     4561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4681 cgcgttgctg gcgtttttcc ataggctccg ccccctgacg agcatcacaa aaatcgacgc
     4741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4861 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     4921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5101 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5281 caagaagatc ctttgttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgtc cgcgcgtt
//
