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
//