LOCUS       40924_2428        7746 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector p15TV-L, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7746)
  AUTHORS   Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C.
  TITLE     Expression vectors for high-throughput In-Fusion cloning
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7746)
  AUTHORS   Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2007) Structural Genomics Consortium, University 
            of Toronto, 100 College Street, Toronto, ON M5G1L5, Canada
REFERENCE   3  (bases 1 to 7746)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7746)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2007) Structural Genomics Consortium, University of Toronto,
            100 College Street, Toronto, ON M5G1L5, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label=AmpR promoter
     CDS             209..1066
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1240..1828
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2014..2156)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2261..2449)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3727..3748)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3764..4843)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4844..4921)
                     /label=lacI promoter
     promoter        5230..5248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /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)."
     RBS             5288..5310
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5329..5346
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             5359..5379
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    complement(5407..5440)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5455..5900
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             5901..7319
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    7395..7409
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     terminator      7487..7534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(7709..7737)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3721 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3841 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3901 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3961 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4021 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     4081 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4141 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4201 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4261 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4321 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4381 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4441 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4501 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4561 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4621 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4681 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4741 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4801 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4861 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4921 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     4981 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     5221 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     5281 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat
     5341 catcacagca gcggcagaga aaacttgtat ttccagggcc atatgagttc tcctcctgaa
     5401 agatccataa cttcgtatag catacattat acgaagttat gcggccgcga cgtccacata
     5461 tacctgccgt tcactattat ttagtgaaat gagatattat gatattttct gaattgtgat
     5521 taaaaaggca actttatgcc catgcaacag aaactataaa aaatacagag aatgaaaaga
     5581 aacagataga ttttttagtt ctttaggccc gtagtctgca aatcctttta tgattttcta
     5641 tcaaacaaaa gaggaaaata gaccagttgc aatccaaacg agagtctaat agaatgaggt
     5701 cgaaaagtaa atcgcgcggg tttgttactg ataaagcagg caagacctaa aatgtgtaaa
     5761 gggcaaagtg tatactttgg cgtcacccct tacatatttt aggtcttttt ttattgtgcg
     5821 taactaactt gccatcttca aacaggaggg ctggaagaag cagaccgcta acacagtaca
     5881 taaaaaagga gacatgaacg atgaacatca aaaagtttgc aaaacaagca acagtattaa
     5941 cctttactac cgcactgctg gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc
     6001 aaaagccata taaggaaaca tacggcattt cccatattac acgccatgat atgctgcaaa
     6061 tccctgaaca gcaaaaaaat gaaaaatatc aagttcctga gttcgattcg tccacaatta
     6121 aaaatatctc ttctgcaaaa ggcctggacg tttgggacag ctggccatta caaaacgctg
     6181 acggcactgt cgcaaactat cacggctacc acatcgtctt tgcattagcc ggagatccta
     6241 aaaatgcgga tgacacatcg atttacatgt tctatcaaaa agtcggcgaa acttctattg
     6301 acagctggaa aaacgctggc cgcgtcttta aagacagcga caaattcgat gcaaatgatt
     6361 ctatcctaaa agaccaaaca caagaatggt caggttcagc cacatttaca tctgacggaa
     6421 aaatccgttt attctacact gatttctccg gtaaacatta cggcaaacaa acactgacaa
     6481 ctgcacaagt taacgtatca gcatcagaca gctctttgaa catcaacggt gtagaggatt
     6541 ataaatcaat ctttgacggt gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg
     6601 aaggcaacta cagctcaggc gacaaccata cgctgagaga tcctcactac gtagaagata
     6661 aaggccacaa atacttagta tttgaagcaa acactggaac tgaagatggc taccaaggcg
     6721 aagaatcttt atttaacaaa gcatactatg gcaaaagcac atcattcttc cgtcaagaaa
     6781 gtcaaaaact tctgcaaagc gataaaaaac gcacggctga gttagcaaac ggcgctctcg
     6841 gtatgattga gctaaacgat gattacacac tgaaaaaagt gatgaaaccg ctgattgcat
     6901 ctaacacagt aacagatgaa attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt
     6961 acctgttcac tgactcccgc ggatcaaaaa tgacgattga cggcattacg tctaacgata
     7021 tttacatgct tggttatgtt tctaattctt taactggccc atacaagccg ctgaacaaaa
     7081 ctggccttgt gttaaaaatg gatcttgatc ctaacgatgt aacctttact tactcacact
     7141 tcgctgtacc tcaagcgaaa ggaaacaatg tcgtgattac aagctatatg acaaacagag
     7201 gattctacgc agacaaacaa tcaacgtttg cgcctagctt cctgctgaac atcaaaggca
     7261 agaaaacatc tgttgtcaaa gacagcatcc ttgaacaagg acaattaaca gttaacaaat
     7321 aaaaacgcaa aagaaaatgc cgatatccta ttggcattga cgtcaggtgg cacttttcga
     7381 ggagatcatg cacatgatga cgaagcttgc ggccgcactc gaggatccgg ctgctaacaa
     7441 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     7501 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     7561 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
     7621 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
     7681 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
     7741 tgagaa
//
