LOCUS       62056_2835        6008 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 6008)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1393)
                     /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(1583..1604)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(1659..1736)
                     /label=lacI promoter
     promoter        1799..1828
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     RBS             1873..1895
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1924..2619
                     /codon_start=1
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     terminator      2988..3074
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3166..3193
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3268..3296
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3344..4531
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(join(5571..6008,1..219))
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctagt ttcagtgcaa
     1441 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     1501 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     1561 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgcctag
     1621 ggccctgcgg agatctgtgt ccggaggtct agaccaccat tcaccaccct gaattgactc
     1681 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     1741 atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactatatt
     1801 gacaattaat catccggctc gtataatggt attttacaca ttagaaaaaa gtttcacaca
     1861 ggaaacagac catttgttta actttaagaa ggagatcagt cagctgaact agtattcgag
     1921 ctcatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     1981 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     2041 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     2101 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac
     2161 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac
     2221 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     2281 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
     2341 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc
     2401 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac
     2461 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga
     2521 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
     2581 gccagatact gcgacctccc tagcaaactg gggcacagag gtacccgggg atcacctgca
     2641 gggcatgctt gacaattaat catccggctc gtataatggg tataacaggt ataaaggtat
     2701 atcccaataa ttttgtttaa ctttaagaag gagactacag tcagaggttg cggccgcatt
     2761 cagctagacg cgtctcaatc tcgagggctg ttttggcgga tgagagaaga ttttcagcct
     2821 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     2881 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     2941 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     3001 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     3061 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     3121 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     3181 cggatggcct ttttttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     3241 atacgatata agttgtaatt ctcatgtttg acagcttatc atcgataagc tttaatgcgg
     3301 tagtttatca cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc
     3361 gctcatcgtc atcctcggca ccgtcaccct ggatgctgta ggcataggct tggttatgcc
     3421 ggtactgccg ggcctcttgc gggatatcgt ccattccgac agcatcgcca gtcactatgg
     3481 cgtgctgcta gcgctatatg cgttgatgca atttctatgc gcacccgttc tcggagcact
     3541 gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg ctacttggag ccactatcga
     3601 ctacgcgatc atggcgacca cacccgtcct gtggatcctc tacgccggac gcatcgtggc
     3661 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     3721 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
     3781 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
     3841 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
     3901 gggagagcgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     3961 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     4021 acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac
     4081 gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt
     4141 cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc
     4201 cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat
     4261 tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag
     4321 gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcct
     4381 aacttcgatc actggaccgc tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg
     4441 gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg
     4501 tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac
     4561 ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa
     4621 accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc
     4681 atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     4741 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     4801 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     4861 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cccctacgtg ctgctgaagt
     4921 tgcccgcaac agagagtgga accaaccggt gataccacga tactatgact gagagtcaac
     4981 gccatgagcg gcctcatttc ttattctgag ttacaacagt ccgcaccgct gtccggtagc
     5041 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     5101 atgcaactcg taggacaggt gccggcagcg cccaacagtc ccccggccac ggggcctgcc
     5161 accataccca cgccgaaaca agcgccctgc accattatgt tccggatctg catcgcagga
     5221 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgcta accgttttta
     5281 tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc acggggagag
     5341 cctgagcaaa ctggcctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     5401 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     5461 cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca
     5521 ggcgtagcac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     5581 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     5641 cagacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     5701 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     5761 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     5821 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     5881 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     5941 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     6001 gccatacg
//
