LOCUS       62056_7835        5913 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5913)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..81
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    84..100
                     /label=lac operator
                     /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)."
     5'UTR           122..255
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     CDS             256..270
                     /codon_start=1
                     /label=TEE
                     /note="translation enhancing element for E. coli (Qing et
                     al., 2004)"
                     /translation="MNHKV"
     CDS             271..288
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             289..300
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             319..1803
                     /codon_start=1
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             1813..1830
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     3'UTR           1874..2018
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(2217..2672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2772..2876
                     /label=AmpR promoter
     CDS             2877..3734
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3908..4496
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(4634..4655)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4671..5750)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5751..5828)
                     /label=lacI promoter
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 cgaaggtagg
      301 catatggagc tcggtaccat gaacaaagaa attttggctg tagttgaagc cgtatccaat
      361 gaaaaggcgc tacctcgcga gaagattttc gaagcattgg aaagcgcgct ggcgacagca
      421 acaaagaaaa aatatgaaca agagatcgac gtccgcgtac agatcgatcg caaaagcggt
      481 gattttgaca ctttccgtcg ctggttagtt gttgatgaag tcacccagcc gaccaaggaa
      541 atcacccttg aagccgcacg ttatgaagat gaaagcctga acctgggcga ttacgttgaa
      601 gatcagattg agtctgttac ctttgaccgt atcactaccc agacggcaaa acaggttatc
      661 gtgcagaaag tgcgtgaagc cgaacgtgcg atggtggttg atcagttccg tgaacacgaa
      721 ggtgaaatca tcaccggcgt ggtgaaaaaa gtaaaccgcg acaacatctc tctggatctg
      781 ggcaacaacg ctgaagccgt gatcctgcgc gaagatatgc tgccgcgtga aaacttccgc
      841 cctggcgacc gcgttcgtgg cgtgctctat tccgttcgcc cggaagcgcg tggcgcgcaa
      901 ctgttcgtca ctcgttccaa gccggaaatg ctgatcgaac tgttccgtat tgaagtgcca
      961 gaaatcggcg aagaagtgat tgaaattaaa gcagcggctc gcgatccggg ttctcgtgcg
     1021 aaaatcgcgg tgaaaaccaa cgataaacgt atcgatccgg taggtgcttg cgtaggtatg
     1081 cgtggcgcgc gtgttcaggc ggtgtctact gaactgggtg gcgagcgtat cgatatcgtc
     1141 ctgtgggatg ataacccggc gcagttcgtg attaacgcaa tggcaccggc agacgttgct
     1201 tctatcgtgg tggatgaaga taaacacacc atggacatcg ccgttgaagc cggtaatctg
     1261 gcgcaggcga ttggccgtaa cggtcagaac gtgcgtctgg cttcgcaact gagcggttgg
     1321 gaactcaacg tgatgaccgt tgacgacctg caagctaagc atcaggcgga agcgcacgca
     1381 gcgatcgaca ccttcaccaa atatctcgac atcgacgaag acttcgcgac tgttctggta
     1441 gaagaaggct tctcgacgct ggaagaattg gcctatgtgc cgatgaaaga gctgttggaa
     1501 atcgaaggcc ttgatgagcc gaccgttgaa gcactgcgcg agcgtgctaa aaatgcactg
     1561 gccaccattg cacaggccca ggaagaaagc ctcggtgata acaaaccggc tgacgatctg
     1621 ctgaaccttg aaggggtaga tcgtgatttg gcattcaaac tggccgcccg tggcgtttgt
     1681 acgctggaag atctcgccga acagggcatt gatgatctgg ctgatatcga agggttgacc
     1741 gacgaaaaag ccggagcact gattatggct gcccgtaata tttgctggtt cggtgacgaa
     1801 gcgggtggaa gtctggtccc ccggggcagc ggatccgaat tcaagcttgt cgacctgcag
     1861 tctagatagg taatctctgc ttaaaagcac agaatctaag atccctgcca tttggcgggg
     1921 atttttttat ttgttttcag gaaataaata atcgatcgcg taataaaatc tattattatt
     1981 tttgtgaaga ataaatttgg gtgcaatgag aatgcgcagg ccctttcgtc tcgcgcgttt
     2041 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2101 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2161 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat
     2221 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2281 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagatagg
     2341 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2401 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     2461 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2521 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2581 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2641 cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt
     2701 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgtcaggt ggcacttttc
     2761 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2821 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2881 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2941 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3001 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3061 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3121 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3181 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3241 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3301 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3361 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3421 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3481 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3541 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3601 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3661 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3721 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4021 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatagtca
     4561 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     4621 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4681 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4741 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4801 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     4861 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4921 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     4981 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     5041 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     5101 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     5161 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     5221 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     5281 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5341 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5401 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5461 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5521 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5581 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     5641 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5701 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5761 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     5821 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5881 tagtaggttg aggccgttga gcaccgccgc cgc
//
