LOCUS       62056_7840        5025 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 5025)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5025
                     /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             850..873
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     CDS             883..900
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             907..918
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     3'UTR           986..1130
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(1329..1784)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1884..1988
                     /label=AmpR promoter
     CDS             1989..2846
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3020..3608
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(3746..3767)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3783..4862)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4863..4940)
                     /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 atcatcacat ggcaaatatt
      301 accgttttct ataacgaaga cttccagggt aagcaggtcg atctgccgcc tggcaactat
      361 acccgcgccc agttggcggc gctgggcatc gagaataata ccatcagctc ggtgaaggtg
      421 ccgcctggcg tgaaggctat cctgtaccag aacgatggtt tcgccggcga ccagatcgaa
      481 gtggtggcca atgccgagga gttgggcccg ctgaataata acgtctccag catccgcgtc
      541 atctccgtgc ccgtgcagcc gcgcatggca aatattaccg ttttctataa cgaagacttc
      601 cagggtaagc aggtcgatct gccgcctggc aactataccc gcgcccagtt ggcggcgctg
      661 ggcatcgaga ataataccat cagctcggtg aaggtgccgc ctggcgtgaa ggctatcctc
      721 taccagaacg atggtttcgc cggcgaccag atcgaagtgg tggccaatgc cgaggagctg
      781 ggtccgctga ataataacgt ctccagcatc cgcgtcatct ccgtgccggt gcagccgagg
      841 tccgcgggtc tggaagttct gttccagggg ccctccgcgg gtctggtgcc acgcggtagt
      901 ggtggtatcg aaggtaggca tatggagctc ggtaccctcg agggatccga attcaagctt
      961 gtcgacctgc agtctagata ggtaatctct gcttaaaagc acagaatcta agatccctgc
     1021 catttggcgg ggattttttt atttgttttc aggaaataaa taatcgatcg cgtaataaaa
     1081 tctattatta tttttgtgaa gaataaattt gggtgcaatg agaatgcgca ggccctttcg
     1141 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1201 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1261 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     1321 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1381 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     1441 gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1501 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1561 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1621 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1681 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     1741 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     1801 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgtcag
     1861 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1921 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1981 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2041 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2101 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2161 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2221 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2281 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2341 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2401 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2461 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2521 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2581 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2641 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2701 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2761 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2821 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2881 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2941 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3661 ctcacatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3721 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3901 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3961 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4021 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     4081 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4141 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     4201 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4261 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4321 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4381 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4441 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4501 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4561 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4621 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4681 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4741 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4801 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4861 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4921 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4981 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgc
//
