LOCUS       40924_16455        4451 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pEamTA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4451)
  AUTHORS   Reisinger C, Kern A, Fesko K, Schwab H.
  TITLE     An efficient plasmid vector for expression cloning of large numbers 
            of PCR fragments in Escherichia coli
  JOURNAL   Appl. Microbiol. Biotechnol. 77 (1), 241-244 (2007)
  PUBMED    17786428
REFERENCE   2  (bases 1 to 4451)
  AUTHORS   Reisinger C, Kern A, Fesko K, Schwab H.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2007) 1 Research Centre Applied Biocatalysis, 
            Petersgasse 14/4, Graz, Styria A-8010, Austria
REFERENCE   3  (bases 1 to 4451)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4451)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2007"; volume: "77"; issue: "1"; 
            pages: "241-244"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2007) 1 Research Centre Applied Biocatalysis, Petersgasse 
            14/4, Graz, Styria A-8010, Austria"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             40..62
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    69
                     /label=single 3'-T overhang in linearized vector
                     /note="single 3'-T overhang in linearized vector"
     misc_feature    517
                     /label=single 3'-T overhang in linearized vector
                     /note="single 3'-T overhang in linearized vector"
     terminator      746..832
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      924..951
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        971..1062
                     /label=AmpR promoter
     CDS             1063..1920
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2094..2682
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2996..4075)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTSKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4076..4153)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        4383..4411
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4419..4435
                     /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)."
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agaaataatt ttgtttaact ttaagaagga
       61 gatgaccata tgtcgacaat ccgacctgac gacaaagcaa tagacgccgc cgcaaggcat
      121 tacggcatca ctctcgacaa aacagcccgg ctcgagtggc cggcactgat cgacggagca
      181 ctgggctcct acgacgtcct ggtgatgcca acgctgccct acgtcgcatc cgaattgccg
      241 gcgaaggacg tagatcgtgc aaccttcatc accaaggctc tcgggatgat cgccaacacg
      301 gcaccattcg acgtgaccgg acatccgtcc ctgtccgttc cggccggcct ggtgaacggg
      361 cttccggtcg gaatgatgat caccggcaga cacttcgacg atgcgacagt ccttcgtgtc
      421 ggacgcgcat tcgaaaagct tcgcggcgcg tttccgacgc cggccgaacg cgcctccaac
      481 tctgcaccac aactcagccc cgcctagtcc tgacgcactg tcagacaaca agcatgcaag
      541 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
      601 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
      661 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
      721 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
      781 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
      841 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
      901 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
      961 acaaactctt tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1021 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1081 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1141 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1201 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1261 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     1321 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1381 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1441 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1501 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1561 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     1621 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1681 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1741 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1801 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg atcccaggca
     1861 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1921 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1981 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2041 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2221 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2461 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     2521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2761 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2821 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     2881 tttctcctta cgcatctgtg cggtatttca caccgcacga acgccagcaa gacgtagccc
     2941 agcgcgtcgg ccagcttgca attcgcgcta acttacatta attgcgttgc gctcactgcc
     3001 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3061 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     3121 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3181 tttgccccag caggcgaaaa tcctgtttgc tggtggttaa cggcgggata taacatgagc
     3241 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     3301 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     3361 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     3421 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     3481 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     3541 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     3601 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     3661 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     3721 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     3781 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     3841 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     3901 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     3961 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4021 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     4081 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc
     4141 attcgatggt gtcaacgtaa atgccgcttc gccttcgcgc gcgaattgca agctgatccg
     4201 ggcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
     4261 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     4321 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     4381 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     4441 cacaggaaac a
//
