LOCUS Exported 5369 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pQTEV- PRDX2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5369) 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 5369) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5369 /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 826..920 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS 964..1623 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1030..1049) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 1688..1774 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(1867..2949) /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 2911..2930 /label=LacI-R /note="LacI, reverse primer" promoter complement(2950..3027) /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(3111..3133) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3219..3359 /label=bom /note="basis of mobility region from pBR322" primer_bind 3274..3293 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3374..3391) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3545..4133) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3625..3644) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4304..5164) /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 4927..4946 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5165..5269) /gene="bla" /label=AmpR promoter primer_bind 5337..5355 /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 cctccggtaa cgcgcgcatc ggaaagccag cccctgactt caaggccaca 241 gcggtggttg atggcgcctt caaagaggtg aagctgtcgg actacaaagg gaagtacgtg 301 gtcctctttt tctaccctct ggacttcacc tttgtgtgcc ccaccgagat catcgcgttc 361 agcaaccgtg cagaggactt ccgcaagctg ggctgtgaag tgctgggcgt ctcggtggac 421 tctcagttca cccacctggc ttggatcaac accccccgga aagagggagg cttgggcccc 481 ctgaacatcc ccctgcttgc tgacgtgacc agacgcttgt ctgaggatta cggcgtgctg 541 aaaacagatg agggcattgc ctacaggggc ctctttatca tcgatggcaa gggtgtcctt 601 cgccagatca ctgttaatga tttgcctgtg ggacgctccg tggatgaggc tctgcggctg 661 gtccaggcct tccagtacac agacgagcat ggggaagttt gtcccgctgg ctggaagcct 721 ggcagtgaca cgattaagcc caacgtggat gacagcaagg aatatttctc caaacacaat 781 taggcggccg ctcgacctgc agccaagctt aattagctga gcttggactc ctgttgatag 841 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg 901 cgttttttat tggtgagaat ccaagctagc ttggcgagat tttcaggagc taaggaagct 961 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa 1021 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg 1081 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt 1141 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac 1201 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact 1261 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata 1321 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt 1381 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac 1441 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa 1501 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc 1561 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg 1621 taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac tctctagctt 1681 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg 1741 tttgtcggtg aacgctctcc tgagtaggac aaatccgccc tctagattac gtgcagtcga 1801 tgataagctg tcaaacatga gaattgtgcc taatgagtga gctaacttac attaattgcg 1861 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 1921 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac 1981 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa 2041 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg 2101 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac 2161 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac 2221 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga 2281 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata 2341 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag 2401 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc 2461 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg 2521 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat 2581 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac 2641 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt 2701 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat 2761 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc 2821 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac 2881 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac 2941 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg 3001 aaaggttttg caccattcga tggtgtcgga atttcgggca gcgttgggtc ctggccacgg 3061 gtgcgcatga tctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 3121 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 3181 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt 3241 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag 3301 tgcaccacat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc 3361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 3421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 3481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 3541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 3601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 3661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 3721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 3781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 3841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 3901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 3961 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 4021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 4141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 4201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 4261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 4321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 4381 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 4441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 4501 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 4561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 4621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 4681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 4741 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 4801 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 4861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 4921 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 4981 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 5041 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 5101 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 5161 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 5221 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 5281 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 5341 ggcgtatcac gaggcccttt cgtcttcac //