LOCUS       62056_7805        4998 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 4998)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4998
                     /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"
     3'UTR           959..1103
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1302..1757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1857..1961
                     /label=AmpR promoter
     CDS             1962..2819
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2993..3581
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3719..3740)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3756..4835)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4836..4913)
                     /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 tcggtacctc tagcggtgct cagtacgaag atggtaaaca gtacactacc
      361 ctggaaaaac cggtagctgg cgcgccgcaa gtgctggagt ttttctcttt cttcagcccg
      421 cattcctatc agtttgaaga agttctgcat atttctgata atgtgaagaa aaaactgccg
      481 gaaggcgtga agatgactaa ataccacgtc aacttcatgg gtggtgacct gggcaaagat
      541 ctgactcagg catgggctgt ggcgatggcg ctgggcgtgg aagacaaagt gactgttccg
      601 ctgtttgaag gcgtacagaa aacccagacc attcgttctg cttctgatat ccgcgatgta
      661 tttatcaacg caggtattaa aggtgaagag tacgacgcgg cgtggaacag cttcgtggtg
      721 aaatctctgg tcgctcagca ggaaaaagct gcagctgacg tgcaattgcg tggcgttccg
      781 gcgatgtttg ttaacggtaa atatcagctg aatccgcagg gtatggatac cagcaatatg
      841 gatgtttttg ttcagcagta tgctgataca gtgaaatatc tgtccgagaa aaaaggtgat
      901 gaagatgaag atagcggatc cgaattcaag cttgtcgacc tgcagtctag ataggtaatc
      961 tctgcttaaa agcacagaat ctaagatccc tgccatttgg cggggatttt tttatttgtt
     1021 ttcaggaaat aaataatcga tcgcgtaata aaatctatta ttatttttgt gaagaataaa
     1081 tttgggtgca atgagaatgc gcaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     1141 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     1201 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     1261 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     1321 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1381 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc
     1441 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1501 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     1561 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1621 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1681 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1741 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     1801 tgcgtaagga gaaaataccg catcaggcgt caggtggcac ttttcgggga aatgtgcgcg
     1861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3601 ttttacggtt cctggccttt tgctggcctt ttgctcacat agtcatgccc cgcgcccacc
     3661 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3721 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3781 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3841 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3901 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3961 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4021 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4081 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     4141 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     4201 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4261 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4321 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4381 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4441 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4501 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4561 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4621 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4681 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4741 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4801 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4861 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     4921 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     4981 gttgagcacc gccgccgc
//
