LOCUS       40924_20721        6510 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pFUSE-HEAVY, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  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  JOURNAL   Unpublished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 6510)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-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 6510)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-NOV-2012) Structural Genomics Consortium, University of Toronto,
            Mars Centre, South Tower, 101 College St, Room 700, Toronto, Ontario
            M5G 1L7, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6510
                     /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     573..632
                     /label=IL-2 signal sequence
                     /note="human interleukin-2 signal sequence"
     misc_feature    633..642
                     /note="partial coding sequence human immunoglobulin heavy
                     chain variable region"
     protein_bind    complement(657..690)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        705..1150
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1151..2569
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    2634..2652
                     /note="partial coding sequence human immunoglobulin heavy
                     chain variable region"
     misc_feature    2634..2648
                     /label=3'-In-Fusion cloning site
                     /note="3'-In-Fusion cloning site"
     misc_feature    2653..2949
                     /note="human immunoglobulin gamma 1 heavy chain constant 1 
                     region"
     misc_feature    2950..2994
                     /label=human immunoglobulin gamma 1 hinge region
                     /note="human immunoglobulin gamma 1 hinge region"
     misc_feature    2995..3324
                     /note="mouse immunoglobulin gamma 1 heavy chain constant 2 
                     region"
     misc_feature    3325..3645
                     /note="mouse immunoglobulin gamma 1 heavy chain constant 3 
                     region"
     polyA_signal    complement(3675..3796)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    complement(3900..4294)
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4311..4682)
                     /label=BleoR
                     /note="antibiotic-binding protein"
     regulatory      complement(4732..4787)
                     /label=EM2KC bacterial promoter
                     /note="EM2KC bacterial promoter"
                     /regulatory_class="promoter"
     regulatory      complement(4905..5167)
                     /label=human ferritin light chain mammalian cell promoter
                     /note="human ferritin light chain mammalian cell promoter"
                     /regulatory_class="promoter"
     enhancer        complement(5275..5578)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5658..6246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    6354..6402
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    6413..6504
                     /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 tcaccggcga aggagggcca ccatgtacag gatgcaactc ctgtcttgca
      601 ttgcactaag tcttgcactt gtcacgaatt cggaggttca gcgctctgaa agatccataa
      661 cttcgtatag catacattat acgaagttat gcggccgcga cgtccacata tacctgccgt
      721 tcactattat ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca
      781 actttatgcc catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga
      841 ttttttagtt ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa
      901 gaggaaaata gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa
      961 atcgcgcggg tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg
     1021 tatactttgg cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt
     1081 gccatcttca aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga
     1141 gacatgaacg atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac
     1201 cgcactgctg gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata
     1261 taaggaaaca tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca
     1321 gcaaaaaaat gaaaaatatc aagttcctga gttcgattcg tccacaatta aaaatatctc
     1381 ttctgcaaaa ggcctggacg tttgggacag ctggccatta caaaacgctg acggcactgt
     1441 cgcaaactat cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga
     1501 tgacacatcg atttacatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa
     1561 aaacgctggc cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa
     1621 agaccaaaca caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt
     1681 attctacact gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt
     1741 taacgtatca gcatcagaca gctctttgaa catcaacggt gtagaggatt ataaatcaat
     1801 ctttgacggt gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta
     1861 cagctcaggc gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa
     1921 atacttagta tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt
     1981 atttaacaaa gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact
     2041 tctgcaaagc gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga
     2101 gctaaacgat gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt
     2161 aacagatgaa attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt acctgttcac
     2221 tgactcccgc ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct
     2281 tggttatgtt tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt
     2341 gttaaaaatg gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc
     2401 tcaagcgaaa ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc
     2461 agacaaacaa tcaacgtttg cgcctagctt cctgctgaac atcaaaggca agaaaacatc
     2521 tgttgtcaaa gacagcatcc ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa
     2581 aagaaaatgc cgatatccta ttggcattga cgtcaggtgg cacttttcgc atgcctggtc
     2641 accgtctcct cggcctccac caagggtcca tcggtcttcc ccctggcacc ctcctccaag
     2701 agcacctctg ggggcacagc ggccctgggc tgcctggtca aggactactt ccccgaaccg
     2761 gtgacggtgt cgtggaactc aggcgccctg accagcggcg tgcacacctt cccggctgtc
     2821 ctacagtcct caggactcta ctccctcagc agcgtggtga ccgtgccctc cagcagcttg
     2881 ggcacccaga cctacatctg caacgtgaat cacaagccca gcaacaccaa ggtcgacaag
     2941 aaagttgagc ccaaatcttg tgacaaaact cacacatgcc ctccatgtcc agcccctgaa
     3001 cttctgggag gaccttctgt cttcatcttc cccccaaagc ccaaggatgt gctcaccatt
     3061 actctgactc ctaaggtcac gtgtgttgtg gtagacatca gcaaggatga tcccgaggtc
     3121 cagttcagct ggtttgtaga tgatgtggag gtgcacacag ctcagacgca accccgggag
     3181 gagcagttca acagcacttt ccgctcagtc agtgaacttc ccatcatgca ccaggactgg
     3241 ctcaatggca aggagttcaa atgcagggtc aacagtgcag ctttccctgc ccccatcgag
     3301 aaaaccatct ccaaaaccaa aggcagaccg aaggctccac aggtgtacac cattccacct
     3361 cccaaggagc agatggccaa ggataaagtc agtctgacct gcatgataac agacttcttc
     3421 cctgaagaca ttactgtgga gtggcagtgg aatgggcagc cagcggagaa ctacaagaac
     3481 actcagccca tcatggacac agatggctct tacttcgtct acagcaagct caatgtgcag
     3541 aagagcaact gggaggcagg aaatactttc acctgctctg tgttacatga gggcctgcac
     3601 aaccaccata ctgagaagag cctctcccac tctcctggta aatgatccca gtgtccctag
     3661 ctggccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3721 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3781 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3841 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg aattaattct
     3901 aaaatacagc atagcaaaac tttaacctcc aaatcaagcc tctacttgaa tccttttctg
     3961 agggatgaat aaggcatagg catcaggggc tgttgccaat gtgcattagc tgtttgcagc
     4021 ctcaccttct ttcatggagt ttaagatata gtgtattttc ccaaggtttg aactagctct
     4081 tcatttcttt atgttttaaa tgcactgacc tcccacattc cctttttagt aaaatattca
     4141 gaaataattt aaatacatca ttgcaatgaa aataaatgtt ttttattagg cagaatccag
     4201 atgctcaagg cccttcataa tatcccccag tttagtagtt ggacttaggg aacaaaggaa
     4261 cctttaatag aaattggaca gcaagaaagc gagcttctag cttatcctca gtcctgctcc
     4321 tctgccacaa agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac
     4381 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg
     4441 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg
     4501 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc
     4561 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac
     4621 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc
     4681 atgatggctc ctcctgtcag gagaggaaag agaagaaggt tagtacaatt gctatagtga
     4741 gttgtattat actatgcaga tatactatgc caatgattaa ttgtcaaact agggctgcag
     4801 ggttcatagt gccacttttc ctgcactgcc ccatctcctg cccacccttt cccaggcata
     4861 gacagtcagt gacttaccaa actcacagga gggagaaggc agaagcttga gacagacccg
     4921 cgggaccgcc gaactgcgag gggacgtggc tagggcggct tcttttatgg tgcgccggcc
     4981 ctcggaggca gggcgctcgg ggaggcctag cggccaatct gcggtggcag gaggcggggc
     5041 cgaaggccgt gcctgaccaa tccggagcac ataggagtct cagccccccg ccccaaagca
     5101 aggggaagtc acgcgcctgt agcgccagcg tgttgtgaaa tgggggcttg ggggggttgg
     5161 ggccctgact agtcaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     5221 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcatcatggt
     5281 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     5341 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     5401 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     5461 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     5521 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcc
     5581 tgcaggttaa ttaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6301 cgttaaggga ttttggtcat ggctagttaa ttaacattta aatcagcggc cgcaataaaa
     6361 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgtaa ctaacatacg
     6421 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     6481 gtgcaggtgc cagaacattt ctctatcgaa
//
