LOCUS       40924_1389        6083 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector HygR-5' EGFP reporter, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  AUTHORS   Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y, 
            Konishi H.
  TITLE     A system for the measurement of gene targeting efficiency in human 
            cell lines using an antibiotic resistance-GFP fusion gene
  JOURNAL   BioTechniques 53 (3), 141-152 (2012)
  PUBMED    22963476
REFERENCE   2  (bases 1 to 6083)
  AUTHORS   Konishi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JAN-2012) Aichi Medical University School of Medicine,
            1-1 Yazako-Karimta, Nagakute, Aichi 480-1195, Japan
REFERENCE   3  (bases 1 to 6083)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6083)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "2012"; volume: "53"; issue: "3"; pages: 
            "141-152"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-JAN-2012) Aichi Medical University School of Medicine, 1-1 
            Yazako-Karimta, Nagakute, Aichi 480-1195, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      936..945
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             957..1973
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     CDS             2025..2531
                     /label=VN173
                     /note="N-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
     polyA_signal    2673..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2865..3883
                     /note="partial SV40 largeT; an internal portion of SV40
                     largeT exon 2"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3950..3966)
                     /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        complement(3974..4004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4019..4040)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4328..4916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5090..5947)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5948..6052)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcctagccac catggataga tccggaaagc
      961 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     1021 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     1081 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     1141 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     1201 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     1261 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     1321 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     1381 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     1441 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     1501 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     1561 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     1621 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     1681 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     1741 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     1801 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     1861 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     1921 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaaccaccac
     1981 caccaccaga ccggtcgtac gtctccggat tcgaattcgc caccatggtg agcaagggcg
     2041 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2101 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2161 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2221 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2281 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2341 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2401 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2461 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2521 tcaactcgag gatccgcggc cgcatagata actgatccag ataactgatc ataatcagcc
     2581 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2641 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2761 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt cgagtgcatt ctagttgtgg
     2821 tttgtccaaa ctcatcaatg tatcttatca tgtctgtact cgagcacaca ggcatagagt
     2881 gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg
     2941 ggttaataag gaatatttga tgtatagtgc cttgactaga gatccatttt ctgttattga
     3001 ggaaagtttg ccaggtgggt taaaggagca tgattttaat ccagaagaag cagaggaaac
     3061 taaacaagtg tcctggaagc ttgtaacaga gtatgcaatg gaaacaaaat gtgatgatgt
     3121 gttgttattg cttgggatgt acttggaatt tcagtacagt tttgaaatgt gtttaaaatg
     3181 tattaaaaaa gaacagccca gccactataa gtaccatgaa aagcattatg caaatgctgc
     3241 tatatttgct gacagcaaaa accaaaaaac catatgccaa caggctgttg atactgtttt
     3301 agctaaaaag cgggttgata gcctacaatt aactagagaa caaatgttaa caaacagatt
     3361 taatgatctt ttggatagga tggatataat gtttggttct acaggctctg ctgacataga
     3421 agaatggatg gctggagttg cttggctaca ctgtttgttg cccaaaatgg attcagtggt
     3481 gtatgacttt ttaaaatgca tggtgtacaa cattcctaaa aaaagatact ggctgtttaa
     3541 aggaccaatt gatagtggta aaactacatt agcagctgct ttgcttgaat tatgtggggg
     3601 gaaagcttta aatgttaatt tgcccttgga caggctgaac tttgagctag gagtagctat
     3661 tgaccagttt ttagtagttt ttgaggatgt aaagggcact ggaggggagt ccagagattt
     3721 gccttcaggt cagggaatta ataacctgga caatttaagg gattatttgg atggcagtgt
     3781 taaggtaaac ttagaaaaga aacacctaaa taaaagaact caaatatttc cccctggaat
     3841 agtcaccatg aatgagtaca gtgtgcctaa aacactgcag gcctcgagta ccgtcgacct
     3901 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3961 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4021 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4081 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4141 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4501 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     4561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4741 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5701 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5761 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5821 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5881 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5941 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6001 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6061 cccgaaaagt gccacctgac gtc
//
