LOCUS       62056_3750        6817 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 6817)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(653..760)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(773..791)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(812..828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(836..852)
                     /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)."
     promoter        complement(860..890)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(905..926)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1214..1802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2022..2124
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2125..2781
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             4541..5563
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     terminator      5736..6303
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus
                     nidulans trpC gene"
     promoter        complement(6689..6794)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
      721 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
      781 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      841 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      961 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1021 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1081 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1141 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1621 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     1681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcgaccgaa
     1921 taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat
     1981 accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt
     2041 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttgtcgaga
     2101 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     2161 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     2221 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     2281 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     2341 ttacgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     2401 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     2461 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     2521 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     2581 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     2641 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     2701 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     2761 gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct
     2821 ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat
     2881 tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt
     2941 ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc
     3001 agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcctt tttttctgat
     3061 caaaagtgct catcattgga aaaccgagat gcgccgcgtg cggctgctgg agatggcgga
     3121 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     3181 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     3241 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     3301 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     3361 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     3421 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     3481 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgag
     3541 ctttgaagtt gctgcaagct ggcttcaagc catcccatcc gaatgtgatg gatgcgttct
     3601 ttctgggccg ttgcgacttt ggggatcgtc tttcccgcgc ccttggttgg aggccctgtc
     3661 tccggtgtcc cttgtccctt ccaggcaagc gagcgaggtc cattcagatg gtgctccatc
     3721 agcgttggct ttccgtctcc attggctctt ggcaattcgg tcagcggggc tgactgcctc
     3781 aggtggggca gtgctagtgt gtgtaccgac ccgcaggatt ggtgctttgc ccagagctct
     3841 acagaatagc gcgcgcatcc atatgttagt tctgcaattt tcttgtatcg gtgctgtgac
     3901 tcatacttcc ccctttggct ggccttgcgg caaccaataa gaacgcacag tgaaatcttg
     3961 cgggtgggga gtggatccat ggcgcctgca ttggcttggg gacgcgcact gtcgcacact
     4021 tccatctgac ctttcagaag ggtttcgtgg tgggcaagga ccaaccggtt gcgcggccgt
     4081 gcgtgggtgc ctcgcccggc actgccaggg ccactgcagt ggcagtttgc tgcctgatac
     4141 aaaatccttc cctccgccca gttttccctc tttgaccttc ctttctcttc tctgcaacca
     4201 aatccaccct atcaaaccaa aacagtatct cgaccgaggt atcaacctga atcagcaaca
     4261 tcgtagccag catttgtctc cgtctctgca gaaccagcga gttgcaaaca ttatccaggc
     4321 aacagggcac caactcactt cttcggcttt caccaatcgg tacagctctt ctcagaactc
     4381 gcgtccgcaa cagttctacg cttcctcagc accttcttca gcttcaatcc tgaacactca
     4441 gaaccgcgca cagcagcgcc ctcctgttcc cttgtttccc aaaagtaccg gtagtatttc
     4501 gcacggaaag cagggcaaca agatgttctc aggtacccat atgaaaaagc ctgaactcac
     4561 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     4621 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     4681 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     4741 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     4801 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     4861 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     4921 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4981 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     5041 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     5101 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     5161 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     5221 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     5281 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     5341 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     5401 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     5461 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     5521 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagagta gatgccgacc
     5581 gggatccact taacgttact gaaatcatca aacagcttga cgaatctgga tataagatcg
     5641 ttggtgtcga tgtcagctcc ggagttgaga caaatggtgt tcaggatctc gataagatac
     5701 gttcatttgt ccaagcagca aagagtgcct tctagtgatt taatagctcc atgtcaacaa
     5761 gaataaaacg cgtttcgggt ttacctcttc cagatacagc tcatctgcaa tgcattaatg
     5821 cattggacct cgcaacccta gtacgccctt caggctccgg cgaagcagaa gaatagctta
     5881 gcagagtcta ttttcatttt cgggagacga gatcaagcag atcaacggtc gtcaagagac
     5941 ctacgagact gaggaatccg ctcttggctc cacgcgacta tatatttgtc tctaattgta
     6001 ctttgacatg ctcctcttct ttactctgat agcttgacta tgaaaattcc gtcaccagcc
     6061 cctgggttcg caaagataat tgcactgttt cttccttgaa ctctcaagcc tacaggacac
     6121 acattcatcg taggtataaa cctcgaaaat cattcctact aagatgggta tacaatagta
     6181 accatggttg cctagtgaat gctccgtaac acccaatacg ccggccgaaa cttttttaca
     6241 actctcctat gagtcgttta cccagaatgc acaggtacac ttgtttagag gtaatccttc
     6301 tttctagcta gaggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     6361 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     6421 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     6481 tgttgggcgc catctccttg cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6721 atcaagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6781 ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
