LOCUS Exported 6311 bp ds-DNA circular SYN 03-MAR-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6311) AUTHORS LS fuer Angewandte Physik LMU - Lukas Milles TITLE Direct Submission JOURNAL Exported Aug 28, 2017 from SnapGene 4.0.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6311 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 12..467 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 167..189 /label=fw to7000thiol CDS complement(560..1375) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 1497..2085 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2515..2706) /codon_start=1 /gene="rop" /product="Rop protein" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3515..4597) /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" promoter complement(4598..4675) /gene="lacI" /label=lacI promoter primer_bind 4960..4979 /label=T7 FW promoter 4984..5002 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 4989..5011 /label=T7FW#2 protein_bind 5003..5027 /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)." primer_bind complement(5015..5074) /label=REV pET28a w/o N Tags HOT primer_bind 5040..5074 /label=#676 FW w/o N-tag pet28a /note="#676 FW w/o N-tag pet28a" primer_bind complement(5043..5073) /label=#349 REV-w/o N-Tags MCS Met /note="#349 REV-w/o N-Tags MCS Met" primer_bind complement(5044..5082) /label=rv ATG to MATyb primer_bind complement(5045..5074) /label=REV N-StrepII MCS primer_bind complement(5045..5074) /label=REV HIS primer_bind complement(5048..5075) /label=REV ybbR primer_bind complement(5049..5080) /label=rv noN-tags MGT primer_bind complement(5049..5080) /label=rv noN-tags1 primer_bind complement(5057..5080) /label=REV-w/o N-Tags MCS RBS 5058..5063 CDS 5072..6163 /codon_start=1 /label=Feature 19 /translation="MGTEQGSNVNHLIKVTDQSITEGYDDSDGIIKAHDAENLIYDVTF EVDDKVKSGDTMTVNIDKNTVPSDLTDSFAIPKIKDNSGEIIATGTYDNTNKQITYTFT DYVDKYENIKAHLKLTSYIDKSKVPNNNTKLDVEYKTALSSVNKTITVEYQKPNENRTA NLQSMFTNIDTKNHTVEQTIYINPLRYSAKETNVNISGNGDEGSTIIDDSTIIKVYKVG DNQNLPDSNRIYDYSEYEDVTNDDYAQLGNNNDVNINFGNIDSPYIIKVISKYDPNKDD YTTIQQTVTMQTTINEYTGEFRTASYDNTIAFSTSSGQGQGDLPPEKTELKLPRSRHHH HHHGSLEVLFQGPDSLEFIASKLA" CDS 5072..5074 /codon_start=1 /label=ATG /translation="M" primer_bind 5077..5098 /label=fw SdrGN2 to y CDS 5081..6058 /codon_start=1 /label=sdrG /translation="EQGSNVNHLIKVTDQSITEGYDDSDGIIKAHDAENLIYDVTFEVD DKVKSGDTMTVNIDKNTVPSDLTDSFAIPKIKDNSGEIIATGTYDNTNKQITYTFTDYV DKYENIKAHLKLTSYIDKSKVPNNNTKLDVEYKTALSSVNKTITVEYQKPNENRTANLQ SMFTNIDTKNHTVEQTIYINPLRYSAKETNVNISGNGDEGSTIIDDSTIIKVYKVGDNQ NLPDSNRIYDYSEYEDVTNDDYAQLGNNNDVNINFGNIDSPYIIKVISKYDPNKDDYTT IQQTVTMQTTINEYTGEFRTASYDNTIAFSTSSGQGQGDLPPEKT" CDS 5087..5533 /codon_start=1 /label=N2 /translation="GSNVNHLIKVTDQSITEGYDDSDGIIKAHDAENLIYDVTFEVDDK VKSGDTMTVNIDKNTVPSDLTDSFAIPKIKDNSGEIIATGTYDNTNKQITYTFTDYVDK YENIKAHLKLTSYIDKSKVPNNNTKLDVEYKTALSSVNKTITVEY" primer_bind complement(5372..5395) /label=rv SdrG L379C misc_feature 5396..5398 /label=L379C primer_bind 5399..5427 /label=fw Sdrg L379C CDS 5534..6052 /codon_start=1 /label=N3 /translation="QKPNENRTANLQSMFTNIDTKNHTVEQTIYINPLRYSAKETNVNI SGNGDEGSTIIDDSTIIKVYKVGDNQNLPDSNRIYDYSEYEDVTNDDYAQLGNNNDVNI NFGNIDSPYIIKVISKYDPNKDDYTTIQQTVTMQTTINEYTGEFRTASYDNTIAFSTSS GQGQGDLPPE" primer_bind complement(5978..6001) /label=rv sdrg nobeta primer_bind complement(6008..6028) /label=rv sdrg truncbeta primer_bind complement(6037..6058) /label=rv QC SdrG B12del CDS 6053..6058 /codon_start=1 /label=sdrG CnA B1 /translation="KT" primer_bind 6057..6081 /label=fw CT link primer_bind complement(6059..6081) /label=REV-CHis zu StrepII primer_bind 6059..6081 /label=FW-C-Tags MCS CDS 6083..6100 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6083..6100) /label=rv HIS to STOP CDS 6107..6130 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 6131..6163 /codon_start=1 /label=ybbR Tag /translation="DSLEFIASKLA" primer_bind 6131..6150 /label=FW ybbR primer_bind 6150..6179 /label=FW QC Cys CTerm primer_bind complement(6164..6187) /label=rv w/o CTags MCS#2 primer_bind 6164..6186 /label=FW-w/o C-Tags MCS primer_bind complement(6164..6181) /label=REV w/o C Tags primer_bind complement(6217..6235) /label=rv to7000 primer_bind complement(6221..6239) /label=T7REV#2 terminator 6239..6286 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(6286..6309) /label=T7 REV ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc 3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa 3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc 3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac 3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca 3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta 3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca 3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa 3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt 3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg 3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca 3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta 3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg 4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat 4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct 4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg 4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat 4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc 4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca 4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg 4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt 4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg 4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct 4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga 4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg 4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc 4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg 4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg 4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga 4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct gtagaaataa 5041 ttttgtttaa ctttaagaag gagatataca tatgggtacc gaacagggta gcaatgtgaa 5101 ccatctgatt aaagttaccg atcagagcat taccgaaggc tatgatgata gtgatggtat 5161 cattaaagca cacgatgccg aaaacctgat ttatgatgtt acctttgagg tggatgataa 5221 agtgaaaagc ggtgatacca tgaccgtgaa cattgataaa aacaccgttc cgagcgatct 5281 gaccgatagc tttgcaattc cgaaaatcaa agataacagc ggtgaaatta ttgccaccgg 5341 cacctatgat aataccaaca aacaaatcac ctacaccttc accgattacg tggataaata 5401 cgaaaacatc aaagcccatc tgaaactgac cagctatatc gataaaagca aagtgccgaa 5461 caacaacacc aaactggatg ttgagtataa aaccgcactg agcagcgtga ataaaaccat 5521 taccgttgaa tatcagaaac cgaacgaaaa tcgtaccgca aatctgcaga gcatgtttac 5581 caatatcgat accaaaaatc ataccgtcga acagaccatt tatatcaatc cgctgcgtta 5641 tagcgccaaa gaaaccaatg tgaatattag cggcaatggt gatgaaggca gcaccattat 5701 tgatgatagc accatcatca aagtgtacaa agtgggcgat aatcagaatc tgccggatag 5761 caatcgcatc tatgattaca gcgaatatga agatgtgacc aacgatgatt atgcacagct 5821 gggtaataat aacgacgtga atatcaactt cggcaatatt gatagcccgt atattatcaa 5881 agtgatcagc aaatacgacc cgaacaaaga tgattacacc accattcagc agaccgttac 5941 catgcagacc accattaatg aatataccgg tgaatttcgt accgccagct atgataacac 6001 aattgcattt agcaccagca gcggtcaggg tcagggtgat ctgcctccgg aaaaaaccga 6061 gctcaagctt cctaggtcta ggcaccacca ccaccaccac ggatccctgg aagttctgtt 6121 ccagggtccg gactctctgg aattcatcgc ttctaaactg gcttaactcg agtaagatcc 6181 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact 6241 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac 6301 tatatccgga t //