LOCUS       62056_7830        5898 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 5898)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5898
                     /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             289..1773
                     /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             1783..1800
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             1825..1842
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     3'UTR           1859..2003
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(2202..2657)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2757..2861
                     /label=AmpR promoter
     CDS             2862..3719
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3893..4481
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(4619..4640)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4656..5735)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5736..5813)
                     /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 catatggagc tcggtaccat gaacaaagaa
      301 attttggctg tagttgaagc cgtatccaat gaaaaggcgc tacctcgcga gaagattttc
      361 gaagcattgg aaagcgcgct ggcgacagca acaaagaaaa aatatgaaca agagatcgac
      421 gtccgcgtac agatcgatcg caaaagcggt gattttgaca ctttccgtcg ctggttagtt
      481 gttgatgaag tcacccagcc gaccaaggaa atcacccttg aagccgcacg ttatgaagat
      541 gaaagcctga acctgggcga ttacgttgaa gatcagattg agtctgttac ctttgaccgt
      601 atcactaccc agacggcaaa acaggttatc gtgcagaaag tgcgtgaagc cgaacgtgcg
      661 atggtggttg atcagttccg tgaacacgaa ggtgaaatca tcaccggcgt ggtgaaaaaa
      721 gtaaaccgcg acaacatctc tctggatctg ggcaacaacg ctgaagccgt gatcctgcgc
      781 gaagatatgc tgccgcgtga aaacttccgc cctggcgacc gcgttcgtgg cgtgctctat
      841 tccgttcgcc cggaagcgcg tggcgcgcaa ctgttcgtca ctcgttccaa gccggaaatg
      901 ctgatcgaac tgttccgtat tgaagtgcca gaaatcggcg aagaagtgat tgaaattaaa
      961 gcagcggctc gcgatccggg ttctcgtgcg aaaatcgcgg tgaaaaccaa cgataaacgt
     1021 atcgatccgg taggtgcttg cgtaggtatg cgtggcgcgc gtgttcaggc ggtgtctact
     1081 gaactgggtg gcgagcgtat cgatatcgtc ctgtgggatg ataacccggc gcagttcgtg
     1141 attaacgcaa tggcaccggc agacgttgct tctatcgtgg tggatgaaga taaacacacc
     1201 atggacatcg ccgttgaagc cggtaatctg gcgcaggcga ttggccgtaa cggtcagaac
     1261 gtgcgtctgg cttcgcaact gagcggttgg gaactcaacg tgatgaccgt tgacgacctg
     1321 caagctaagc atcaggcgga agcgcacgca gcgatcgaca ccttcaccaa atatctcgac
     1381 atcgacgaag acttcgcgac tgttctggta gaagaaggct tctcgacgct ggaagaattg
     1441 gcctatgtgc cgatgaaaga gctgttggaa atcgaaggcc ttgatgagcc gaccgttgaa
     1501 gcactgcgcg agcgtgctaa aaatgcactg gccaccattg cacaggccca ggaagaaagc
     1561 ctcggtgata acaaaccggc tgacgatctg ctgaaccttg aaggggtaga tcgtgatttg
     1621 gcattcaaac tggccgcccg tggcgtttgt acgctggaag atctcgccga acagggcatt
     1681 gatgatctgg ctgatatcga agggttgacc gacgaaaaag ccggagcact gattatggct
     1741 gcccgtaata tttgctggtt cggtgacgaa gcgggtggaa gtctggtccc ccggggcagc
     1801 ggatccgaat tcaagcttgt cgaccatcat catcatcatc actaatctag ataggtaatc
     1861 tctgcttaaa agcacagaat ctaagatccc tgccatttgg cggggatttt tttatttgtt
     1921 ttcaggaaat aaataatcga tcgcgtaata aaatctatta ttatttttgt gaagaataaa
     1981 tttgggtgca atgagaatgc gcaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2041 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2101 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2161 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     2221 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2281 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc
     2341 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2401 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     2461 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2521 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2581 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2641 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     2701 tgcgtaagga gaaaataccg catcaggcgt caggtggcac ttttcgggga aatgtgcgcg
     2761 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4501 ttttacggtt cctggccttt tgctggcctt ttgctcacat agtcatgccc cgcgcccacc
     4561 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4621 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4681 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4741 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4801 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4861 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4921 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4981 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5041 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5101 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     5161 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     5221 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     5281 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5341 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5401 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     5461 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     5521 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5581 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     5641 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5701 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5761 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     5821 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     5881 gttgagcacc gccgccgc
//
