LOCUS       Exported                5453 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pQTEV-PSMD10
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  AUTHORS   Bussow K, Scheich C, Sievert V, Harttig U, Schultz J, Simon B, Bork 
            P, Lehrach H, Heinemann U
  TITLE     Structural genomics of human proteins--target selection and 
            generation of a public catalogue of expression clones.
  JOURNAL   Microb Cell Fact. 2005 Jul 5. 4():21.
  PUBMED    15998469
REFERENCE   2  (bases 1 to 5453)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5453
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             121..141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             166..186
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      910..1004
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1048..1707
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1114..1133)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1772..1858
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1951..3033)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2995..3014
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3034..3111)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3195..3217)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3303..3443
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3358..3377
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3458..3475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3629..4217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3709..3728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4388..5248)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5011..5030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5249..5353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5421..5439
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
      121 catcaccatc accatcacca tagcgattac gacatcccca ctactgagaa tctttatttt
      181 cagggatccg aggggtgtgt gtctaaccta atggtctgca acctggccta cagcgggaag
      241 ctggaagagt tgaaggagag tattctggcc gataaatccc tggctactag aactgaccag
      301 gacagcagaa ctgcattgca ctgggcatgc tcagctggac atacagaaat tgttgaattt
      361 ttgttgcaac ttggagtgcc agtgaatgat aaagacgatg caggttggtc tcctcttcat
      421 attgcggctt ctgctggccg ggatgagatt gtaaaagccc ttctgggaaa aggtgctcaa
      481 gtgaatgctg tcaatcaaaa tggctgtact cccttacatt atgcagcttc gaaaaacagg
      541 catgagatcg ctgtcatgtt actggaaggc ggggctaatc cagatgctaa ggaccattat
      601 gaggctacag caatgcaccg ggcagcagcc aagggtaact tgaagatgat tcatatcctt
      661 ctgtactaca aagcatccac aaacatccaa gacactgagg gtaacactcc tctacactta
      721 gcctgtgatg aggagagagt ggaagaagca aaactgctgg tgtcccaagg agcaagtatt
      781 tacattgaga ataaagaaga aaagacaccc ctgcaagtgg ccaaaggtgg cctgggttta
      841 atactcaaga gaatggtgga aggttaagcg gccgctcgac ctgcagccaa gcttaattag
      901 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
      961 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg
     1021 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1081 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1141 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1201 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1261 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1321 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1381 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1441 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1501 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1561 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1621 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1681 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     1741 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1801 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1861 gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg tgcctaatga
     1921 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1981 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2041 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     2101 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     2161 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     2221 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     2281 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     2341 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     2401 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     2461 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     2521 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     2581 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     2641 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     2701 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     2761 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     2821 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     2881 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     2941 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     3001 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     3061 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cggaatttcg
     3121 ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc gcgtttcggt
     3181 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3241 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3301 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     3361 catcagagca gattgtactg agagtgcacc acatgcggtg tgaaataccg cacagatgcg
     3421 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4141 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4201 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4261 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4321 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4381 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4441 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4501 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4561 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4621 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4681 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4741 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4801 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4861 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4921 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4981 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5041 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5101 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5161 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5221 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5281 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5341 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5401 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
