LOCUS       62056_18705        4349 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 4349)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    21..45
                     /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)."
     RBS             60..82
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             104..130
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     CDS             134..154
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             723..767
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      819..866
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1090..1746)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1747..1849)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(2375..2920)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(3111..3132)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3148..4227)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4228..4305)
                     /label=lacI promoter
ORIGIN
        1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt
       61 ttgtttaact ttaataagga gatataccat ggcatcgtac taccatcacc atcaccatca
      121 ccatcaccat cacgaaaacc tgtattttca gggcagcaac aggagactgc agcagaccca
      181 ggcccaggtg gatgaggtgg tggacatcat gagggtgaat gtggacaagg tcctggagcg
      241 agaccagaag ctatcggaac tggatgatcg ctaggatgcc ctccaggcag gggcctccca
      301 gtttgaaaca agtgcagcca agctcaagcg caaatactgg tagctcgaca agcttgcggc
      361 cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat
      421 taatacgact cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta
      481 gttaagtata agaaggagat atacatatgg ccctcagtga gatcgagacc aggcacagtg
      541 agatcatcaa gttggagaac agcatccggg agctacacga tatgttcatg gacatggcca
      601 tgctggtgga gagccagggg gagatgattg acaggatcga gtacaatgtg gaacacgctg
      661 tggactacgt ggagagggcc gtgtctgaca ccaagaaggc cgtcaagtaa ctcgagtctg
      721 gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag
      781 cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      841 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
      901 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
      961 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     1021 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     1081 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1141 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1201 cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat
     1261 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1321 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1381 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1441 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1501 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag
     1561 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1621 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1681 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1741 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1801 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     1861 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     1921 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     1981 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     2041 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     2101 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     2161 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     2221 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     2281 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     2341 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     2401 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     2461 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     2521 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     2581 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     2641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     2701 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     2761 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     2821 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     2881 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     2941 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     3001 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     3061 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     3121 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3181 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     3241 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     3301 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     3361 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     3421 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     3481 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     3541 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     3601 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     3661 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     3721 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     3781 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     3841 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     3901 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     3961 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     4021 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     4081 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     4141 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     4201 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     4261 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     4321 ctcccttatg cgactcctgc attaggaaa
//
