LOCUS pDEST-ColdII 6600 bp DNA circular 30-APR-2014 FEATURES Location/Qualifiers source join(2560..6600,1..294) /dnas_title="derived form pColdII" /organism="Expression vector pColdII" /mol_type="other DNA" /db_xref="taxon:289230" /lab_host="Escherichia coli" /note="expresses an open reading frame as a fusion to a His6 containing translation enhancing element. This plasmid is derived from pUC118." promoter 15..81 /gene="cspA" misc_feature 82..121 /gene="lacO" /note="synthetic lac operator sequence" 5'UTR 122..255 /gene="modified cspA 5'UTR" /note="modified cspA 5'UTR" misc_feature 256..270 /gene="TEE" /note="TEE (translation enhancing element)" misc_feature 271..288 /gene="His6" /note="His6 tag" 3'UTR 2561..2705 /gene="cspA 3'UTR" /note="cspA 3'UTR" misc_feature complement(2904..3377) /note="M13 intergenic region" gene 3564..4424 /gene="Amp" CDS 3564..4424 /dnas_title="beta-lactamase" /gene="Amp" /codon_start=1 /transl_table=11 /product="beta-lactamase" /protein_id="BAD35136.1" /db_xref="GI:51090251" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 4584..5198 /note="ColE1 origin" gene complement(5355..6437) /gene="lacI" CDS complement(5355..6437) /dnas_title="lactose operon repressor" /gene="lacI" /codon_start=1 /transl_table=11 /product="lactose operon repressor" /protein_id="BAD35137.1" /db_xref="GI:51090252" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT QTASPRALADSLMQLARQVSRLESGQ" misc_recomb 295..419 /note="attR1" misc_recomb 303..309 /note="attR1 break" CDS complement(848..1153) /gene="ccdB" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSD KVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" CDS complement(1495..2154) /note="Cm resistance" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG A" misc_recomb 2435..2559 /note="attR2" misc_recomb 2545..2551 /note="attR2 break" ORIGIN 1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa 61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat 121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt 181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt 241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat ggacACAAGT 301 TTGTACAAAA AAGCTGAACG AGAAACGTAA AATGATATAA ATATCAATAT ATTAAATTAG 361 ATTTTGCATA AAAAACAGAC TACATAATAC TGTAAAACAC AACATATCCA GTCACTATGA 421 ATCAACTACT TAGATGGTAT TAGTGACCTG TAGTCGACCG ACAGCCTTCC AAATGTTCTT 481 CGGGTGATGC TGCCAACTTA GTCGACCGAC AGCCTTCCAA ATGTTCTTCT CAAACGGAAT 541 CGTCGTATCC AGCCTACTCG CTATTGTCCT CAATGCCGTA TTAAATCATA AAAAGAAATA 601 AGAAAAAGAG GTGCGAGCCT CTTTTTTGTG TGACAAAATA AAAACATCTA CCTATTCATA 661 TACGCTAGTG TCATAGTCCT GAAAATCATC TGCATCAAGA ACAATTTCAC AACTCTTATA 721 CTTTTCTCTT ACAAGTCGTT CGGCTTCATC TGGATTTTCA GCCTCTATAC TTACTAAACG 781 TGATAAAGTT TCTGTAATTT CTACTGTATC GACCTGCAGA CTGGCTGTGT ATAAGGGAGC 841 CTGACATTTA TATTCCCCAG AACATCAGGT TAATGGCGTT TTTGATGTCA TTTTCGCGGT 901 GGCTGAGATC AGCCACTTCT TCCCCGATAA CGGAGACCGG CACACTGGCC ATATCGGTGG 961 TCATCATGCG CCAGCTTTCA TCCCCGATAT GCACCACCGG GTAAAGTTCA CGGGAGACTT 1021 TATCTGACAG CAGACGTGCA CTGGCCAGGG GGATCACCAT CCGTCGCCCG GGCGTGTCAA 1081 TAATATCACT CTGTACATCC ACAAACAGAC GATAACGGCT CTCTCTTTTA TAGGTGTAAA 1141 CCTTAAACTG CATTTCACCA GTCCCTGTTC TCGTCAGCAA AAGAGCCGTT CATTTCAATA 1201 AACCGGGCGA CCTCAGCCAT CCCTTCCTGA TTTTCCGCTT TCCAGCGTTC GGCACGCAGA 1261 CGACGGGCTT CATTCTGCAT GGTTGTGCTT ACCAGACCGG AGATATTGAC ATCATATATG 1321 CCTTGAGCAA CTGATAGCTG TCGCTGTCAA CTGTCACTGT AATACGCTGC TTCATAGCAC 1381 ACCTCTTTTT GACATACTTC GGGTATACAT ATCAGTATAT ATTCTTATAC CGCAAAAATC 1441 AGCGCGCAAA TACGCATACT GTTATCTGGC TTTTAGTAAG CCGGATCCAC GCGATTACGC 1501 CCCGCCCTGC CACTCATCGC AGTACTGTTG TAATTCATTA AGCATTCTGC CGACATGGAA 1561 GCCATCACAG ACGGCATGAT GAACCTGAAT CGCCAGCGGC ATCAGCACCT TGTCGCCTTG 1621 CGTATAATAT TTGCCCATGG TGAAAACGGG GGCGAAGAAG TTGTCCATAT TGGCCACGTT 1681 TAAATCAAAA CTGGTGAAAC TCACCCAGGG ATTGGCTGAG ACGAAAAACA TATTCTCAAT 1741 AAACCCTTTA GGGAAATAGG CCAGGTTTTC ACCGTAACAC GCCACATCTT GCGAATATAT 1801 GTGTAGAAAC TGCCGGAAAT CGTCGTGGTA TTCACTCCAG AGCGATGAAA ACGTTTCAGT 1861 TTGCTCATGG AAAACGGTGT AACAAGGGTG AACACTATCC CATATCACCA GCTCACCGTC 1921 TTTCATTGCC ATACGGAATT CCGGATGAGC ATTCATCAGG CGGGCAAGAA TGTGAATAAA 1981 GGCCGGATAA AACTTGTGCT TATTTTTCTT TACGGTCTTT AAAAAGGCCG TAATATCCAG 2041 CTGAACGGTC TGGTTATAGG TACATTGAGC AACTGACTGA AATGCCTCAA AATGTTCTTT 2101 ACGATGCCAT TGGGATATAT CAACGGTGGT ATATCCAGTG ATTTTTTTCT CCATTTTAGC 2161 TTCCTTAGCT CCTGAAAATC TCGATAACTC AAAAAATACG CCCGGTAGTG ATCTTATTTC 2221 ATTATGGTGA AAGTTGGAAC CTCTTACGTG CCGATCAACG TCTCATTTTC GCCAAAAGTT 2281 GGCCCAGGGC TTCCCGGTAT CAACAGGGAC ACCAGGATTT ATTTATTCTG CGAAGTGATC 2341 TTCCGTCACA GGTATTTATT CGGCGCAAAG TGCGTCGGGT GATGCTGCCA ACTTAGTCGA 2401 CTACAGGTCA CTAATACCAT CTAAGTAGTT GATTCATAGT GACTGGATAT GTTGTGTTTT 2461 ACAGTATTAT GTAGTCTGTT TTTTATGCAA AATCTAATTT AATATATTGA TATTTATATC 2521 ATTTTACGTT TCTCGTTCAG CTTTCTTGTA CAAAGTGGTc tctctgctta aaagcacaga 2581 atctaagatc cctgccattt ggcggggatt tttttatttg ttttcaggaa ataaataatc 2641 gatcgcgtaa taaaatctat tattattttt gtgaagaata aatttgggtg caatgagaat 2701 gcgcaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 2761 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 2821 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca 2881 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt 2941 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta 3001 taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 3061 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 3121 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact 3181 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 3241 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 3301 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta 3361 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac 3421 cgcatcaggc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 3481 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 3541 aataatattg aaaaaggaag agtATGAGTA TTCAACATTT CCGTGTCGCC CTTATTCCCT 3601 TTTTTGCGGC ATTTTGCCTT CCTGTTTTTG CTCACCCAGA AACGCTGGTG AAAGTAAAAG 3661 ATGCTGAAGA TCAGTTGGGT GCACGAGTGG GTTACATCGA ACTGGATCTC AACAGCGGTA 3721 AGATCCTTGA GAGTTTTCGC CCCGAAGAAC GTTTTCCAAT GATGAGCACT TTTAAAGTTC 3781 TGCTATGTGG CGCGGTATTA TCCCGTATTG ACGCCGGGCA AGAGCAACTC GGTCGCCGCA 3841 TACACTATTC TCAGAATGAC TTGGTTGAGT ACTCACCAGT CACAGAAAAG CATCTTACGG 3901 ATGGCATGAC AGTAAGAGAA TTATGCAGTG CTGCCATAAC CATGAGTGAT AACACTGCGG 3961 CCAACTTACT TCTGACAACG ATCGGAGGAC CGAAGGAGCT AACCGCTTTT TTGCACAACA 4021 TGGGGGATCA TGTAACTCGC CTTGATCGTT GGGAACCGGA GCTGAATGAA GCCATACCAA 4081 ACGACGAGCG TGACACCACG ATGCCTGTAG CAATGGCAAC AACGTTGCGC AAACTATTAA 4141 CTGGCGAACT ACTTACTCTA GCTTCCCGGC AACAATTAAT AGACTGGATG GAGGCGGATA 4201 AAGTTGCAGG ACCACTTCTG CGCTCGGCCC TTCCGGCTGG CTGGTTTATT GCTGATAAAT 4261 CTGGAGCCGG TGAGCGTGGG TCTCGCGGTA TCATTGCAGC ACTGGGGCCA GATGGTAAGC 4321 CCTCCCGTAT CGTAGTTATC TACACGACGG GGAGTCAGGC AACTATGGAT GAACGAAATA 4381 GACAGATCGC TGAGATAGGT GCCTCACTGA TTAAGCATTG GTAActgtca gaccaagttt 4441 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 4501 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 4561 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 4621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 4681 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 4741 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 4801 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 4861 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 4921 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 4981 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 5041 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 5101 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 5161 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 5221 tttgctggcc ttttgctcac atagtcatgc cccgcgccca ccggaaggag ctgactgggt 5281 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat 5341 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5401 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt 5461 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt 5521 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt 5581 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc 5641 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg 5701 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga 5761 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga 5821 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc 5881 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta 5941 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat 6001 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga 6061 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag 6121 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg 6181 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca 6241 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa 6301 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc 6361 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat 6421 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc 6481 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg 6541 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc //