LOCUS       40924_4994        3828 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pATE, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3828)
  AUTHORS   Buth SA, Shneider MM, Scholl D, Leiman PG.
  TITLE     Structure and Analysis of R1 and R2 Pyocin Receptor-Binding Fibers
  JOURNAL   Viruses 10 (8), E427 (2018)
  PUBMED    30110933
REFERENCE   2  (bases 1 to 3828)
  AUTHORS   Shneider M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUL-2018) Laboratory of Molecular Engineering, IBCH 
            RAN, Miklukho-Maklaya str, Moscow 117997, Russia
REFERENCE   3  (bases 1 to 3828)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3828)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Viruses"; 
            date: "2018"; volume: "10"; issue: "8"; pages: "E427"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUL-2018) Laboratory of Molecular Engineering, IBCH RAN, 
            Miklukho-Maklaya str, Moscow 117997, Russia"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             79..101
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             120..137
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             147..167
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    171..217
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             223..267
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      319..366
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(589..1245)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1246..1348)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1874..2418)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(2609..2630)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2646..3725)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3726..3803)
                     /label=lacI promoter
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atatcccatg ggcagcagcc
      121 atcatcatca tcatcacagc agcggcgaga atctgtattt ccagggcagc ggatccgaat
      181 tcgagctccg tcgacaagct tgcggccgca ctcgagtctg gtaaagaaac cgctgctgcg
      241 aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct
      301 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      361 tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
      421 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
      481 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
      541 cgtagcacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
      601 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
      661 gacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
      721 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
      781 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
      841 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
      901 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
      961 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     1021 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     1081 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     1141 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     1201 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     1261 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     1321 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     1381 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     1441 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat
     1501 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag
     1561 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
     1621 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     1681 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     1741 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     1801 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     1861 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     1921 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt cccctggcgg
     1981 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     2041 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     2101 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     2161 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     2221 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     2281 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     2341 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     2401 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca
     2461 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     2521 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     2581 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     2641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2701 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     2761 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     2821 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     2881 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     2941 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     3001 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     3061 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     3121 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     3181 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     3241 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     3301 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     3361 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     3421 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     3481 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     3541 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     3601 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     3661 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     3721 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     3781 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatg
//
