LOCUS       pQTEV-STX10                5513 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pQTEV-STX10
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  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 5513)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..5513
                     /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      970..1064
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1108..1767
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1174..1193)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1832..1918
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2011..3093)
                     /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     3055..3074
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3094..3171)
                     /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(3255..3277)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3363..3503
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3418..3437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3518..3535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3689..4277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3769..3788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4448..5308)
                     /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     5071..5090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5309..5413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5481..5499
                     /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 cagggatcct ctctcgaaga cccctttttt gtagtccgag gcgaggtgca gaaggcggtg
      241 aacacggccc gcgggctgta ccagcgctgg tgcgagctcc tgcaggaaag cgcggcggtc
      301 ggacgcgagg agctggactg gacgaccaat gagctgcgga atggcctgcg cagcatcgag
      361 tgggacctcg aggacctgga agagaccatc ggtatagtgg aagccaaccc aggcaagttc
      421 aagctcccag ccggggacct gcaggagaga aaggtgttcg tggagcggat gcgagaggca
      481 gtccaggaaa tgaaggacca tatggtcagc ccaacagccg tagcattttt ggagaggaat
      541 aacagagaga tactcgcagg caagccagct gcccagaagt cacccagcga cctgctggat
      601 gccagcgcag tctcggccac atctcgctac atcgaggagc agcaggccac acagcagctg
      661 atcatggatg aacaggatca acagctggag atggtgtctg ggagcatcca ggttctgaag
      721 cacatgtccg gccgcgttgg agaagagctg gacgagcagg gcatcatgct ggatgccttc
      781 gcccaagaga tggaccacac ccagtcccgc atggacgggg tcctcaggaa gttggccaaa
      841 gtatcccaca tgacgagtgg tgagtcccct caggggaggg gtcagtcctg gtgggggcag
      901 gttgtaggtg gtaccctctc cccgtgagcg gccgctcgac ctgcagccaa gcttaattag
      961 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1021 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg
     1081 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1141 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1201 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1261 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1321 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1381 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1441 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1501 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1561 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1621 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1681 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1741 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     1801 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1861 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1921 gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg tgcctaatga
     1981 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2041 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2101 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     2161 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     2221 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     2281 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     2341 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     2401 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     2461 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     2521 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     2581 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     2641 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     2701 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     2761 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     2821 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     2881 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     2941 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     3001 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     3061 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     3121 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cggaatttcg
     3181 ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc gcgtttcggt
     3241 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3301 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3361 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     3421 catcagagca gattgtactg agagtgcacc acatgcggtg tgaaataccg cacagatgcg
     3481 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//