LOCUS       V006151                 5983 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006151
VERSION     V006151
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5983)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     A pipeline to generate recombinant antibodies to human target
            proteins involved in epigenetic mechanisms
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5983)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2012) Structural Genomics Consortium, University
            of Toronto, Mars Centre, South Tower, 101 College St, Room 700,
            Toronto, Ontario M5G 1L7, Canada
REFERENCE   3  (bases 1 to 5983)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5983)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-NOV-2012) Structural Genomics Consortium, University of Toronto,
            Mars Centre, South Tower, 101 College St, Room 700, Toronto, Ontario
            M5G 1L7, Canada"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..242
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             255..523
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     sig_peptide     556..615
                     /label="IL-2 signal sequence"
                     /note="human interleukin-2 signal sequence"
     misc_feature    616..753
                     /note="partial coding sequence human Immunoglobulin light
                     chain, kappa type"
     misc_feature    694..708
                     /label="5'-In-Fusion cloning site"
                     /note="5'-In-Fusion cloning site"
     protein_bind    complement(765..798)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        813..1258
                     /label="sacB promoter"
                     /note="sacB promoter and control region"
     CDS             1259..2677
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
     misc_feature    2737..2757
                     /note="partial coding sequence human Immunoglobulin light
                     chain, kappa type"
     misc_feature    2742..2757
                     /label="3'-In-Fusion cloning site"
                     /note="3'-In-Fusion cloning site"
     CDS             2761..3078
                     /label="hIg-kappa-CL"
                     /note="Human immunoglobulin kappa light chain constant
                     region"
     polyA_signal    complement(3118..3239)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     polyA_signal    complement(3343..3737)
                     /label="beta-globin poly(A)"
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(3749..4168)
                     /gene="bsr"
                     /label="Blasticidin-S deaminase"
                     /note="Blasticidin-S deaminase from Bacillus cereus.
                     Accession#: P33967"
     regulatory      complement(4218..4260)
                     /label="EM2KC bacterial promoter"
                     /note="EM2KC bacterial promoter"
                     /regulatory_class="promoter"
     regulatory      complement(4378..4640)
                     /label="human ferritin light chain mammalian cell promoter"
                     /note="human ferritin light chain mammalian cell promoter"
                     /regulatory_class="promoter"
     enhancer        complement(4748..5051)
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     polyA_signal    5827..5875
                     /label="poly(A) signal"
                     /note="synthetic polyadenylation signal"
     misc_feature    5886..5977
                     /label="pause site"
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
       61 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
      121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      241 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
      301 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
      361 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
      421 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
      481 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
      541 ctacctgaga tcaacatgta caggatgcaa ctcctgtctt gcattgcact aagtcttgca
      601 cttgtcacga attcagatat ccagatgacc cagtccccga gctccctgtc cgcctctgtg
      661 ggcgataggg tcaccatcac ctgccgtgcc agtcagtccg tgtccagcgc tgtagcctgg
      721 tatcaacaga aaccaggaaa agctccgaag cttctgaaag atccataact tcgtatagca
      781 tacattatac gaagttatgc ggccgcgacg tccacatata cctgccgttc actattattt
      841 agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac tttatgccca
      901 tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt ttttagttct
      961 ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga ggaaaataga
     1021 ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt
     1081 tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg
     1141 tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa
     1201 caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgat
     1261 gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg cactgctggc
     1321 aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata aggaaacata
     1381 cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc aaaaaaatga
     1441 aaaatatcaa gttcctgagt tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg
     1501 cctggacgtt tgggacagct ggccattaca aaacgctgac ggcactgtcg caaactatca
     1561 cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg acacatcgat
     1621 ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa acgctggccg
     1681 cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag accaaacaca
     1741 agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat tctacactga
     1801 tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta acgtatcagc
     1861 atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct ttgacggtga
     1921 cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca gctcaggcga
     1981 caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat acttagtatt
     2041 tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat ttaacaaagc
     2101 atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc tgcaaagcga
     2161 taaaaaacgc acggctgagt tagcaaacgg cgctctcggt atgattgagc taaacgatga
     2221 ttacacactg aaaaaagtga tgaaaccgct gattgcatct aacacagtaa cagatgaaat
     2281 tgaacgcgcg aacgtcttta aaatgaacgg caaatggtac ctgttcactg actcccgcgg
     2341 atcaaaaatg acgattgacg gcattacgtc taacgatatt tacatgcttg gttatgtttc
     2401 taattcttta actggcccat acaagccgct gaacaaaact ggccttgtgt taaaaatgga
     2461 tcttgatcct aacgatgtaa cctttactta ctcacacttc gctgtacctc aagcgaaagg
     2521 aaacaatgtc gtgattacaa gctatatgac aaacagagga ttctacgcag acaaacaatc
     2581 aacgtttgcg cctagcttcc tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga
     2641 cagcatcctt gaacaaggac aattaacagt taacaaataa aaacgcaaaa gaaaatgccg
     2701 atatcctatt ggcattgacg tcaggtggca cttttcggta ccaaggtgga gatcaaacga
     2761 actgtggctg caccatctgt cttcatcttc ccgccatctg attcacagtt gaaatctgga
     2821 actgcctctg ttgtgtgcct gctgaataac ttctatccca gagaggccaa agtacagtgg
     2881 aaggtggata acgccctcca atcgggtaac tcccaggaga gtgtcacaga gcaggacagc
     2941 aaggacagca cctacagcct cagcagcacc ctgacgctga gcaaagcaga ctacgaaaaa
     3001 cataaagtct acgcctgcga agtcacccat cagggcctga gctcgcccgt cacaaagagc
     3061 ttcaacaggg gagagtgtta gagacaaagg tcctgagagc tagctggcca gacatgataa
     3121 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3181 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3241 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     3301 aaagcaagta aaacctctac aaatgtggta tggaattaat tctaaaatac agcatagcaa
     3361 aactttaacc tccaaatcaa gcctctactt gaatcctttt ctgagggatg aataaggcat
     3421 aggcatcagg ggctgttgcc aatgtgcatt agctgtttgc agcctcacct tctttcatgg
     3481 agtttaagat atagtgtatt ttcccaaggt ttgaactagc tcttcatttc tttatgtttt
     3541 aaatgcactg acctcccaca ttcccttttt agtaaaatat tcagaaataa tttaaataca
     3601 tcattgcaat gaaaataaat gttttttatt aggcagaatc cagatgctca aggcccttca
     3661 taatatcccc cagtttagta gttggactta gggaacaaag gaacctttaa tagaaattgg
     3721 acagcaagaa agcgagcttc tagctttagt tcctggtgta cttgaggggg atgagttcct
     3781 caatggtggt tttgaccagc ttgccattca tctcaatgag cacaaagcag tcaggagcat
     3841 agtcagagat gagctctctg cacatgccac aggggctgac caccctgatg gatctgtcca
     3901 cctcatcaga gtaggggtgc ctgacagcca caatggtgtc aaagtccttc tgcccgttgc
     3961 tcacagcaga cccaatggca atggcttcag cacagacagt gaccctgcca atgtaggcct
     4021 caatgtggac agcagagatg atctccccag tcttggtcct gatggccgcc ccgacatggt
     4081 gcttgttgtc ctcatagagc atggtgatct tctcagtggc gacctccacc agctccagat
     4141 cctgctgaga gatgttgaag gtcttcatga tggctcctcc tgtcaggaga ggaaagagaa
     4201 gaaggttagt acaattgcta tagtgagttg tattatacta tgcttatgat taattgtcaa
     4261 actagggctg cagggttcat agtgccactt ttcctgcact gccccatctc ctgcccaccc
     4321 tttcccaggc atagacagtc agtgacttac caaactcaca ggagggagaa ggcagaagct
     4381 tgagacagac ccgcgggacc gccgaactgc gaggggacgt ggctagggcg gcttctttta
     4441 tggtgcgccg gccctcggag gcagggcgct cggggaggcc tagcggccaa tctgcggtgg
     4501 caggaggcgg ggccgaaggc cgtgcctgac caatccggag cacataggag tctcagcccc
     4561 ccgccccaaa gcaaggggaa gtcacgcgcc tgtagcgcca gcgtgttgtg aaatgggggc
     4621 ttgggggggt tggggccctg actagtcaaa acaaactccc attgacgtca atggggtgga
     4681 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     4741 gcatcatcat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4801 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4861 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4921 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4981 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     5041 gttatgtaac gcctgcaggt taattaagaa catgtgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5761 gaacgaaaac tcacgttaag ggattttggt catggctagt taattaacat ttaaatcagc
     5821 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     5881 taactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     5941 gtccccagtg caagtgcagg tgccagaaca tttctctatc gaa
//
