LOCUS       62056_17175        7049 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 7049)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    358..457
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    496..544
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    558..649
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     CDS             789..2438
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        2519..2848
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2911..3843
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    3862..4448
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4468..5064
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    complement(5167..5288)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5500..6306
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6399..6987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttcg aaggagatag aaccagatct
      481 tggaattcag gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
      541 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
      601 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta ctagtacgcg
      661 tcctcaggtc tcccagtctt ttactgggct gatcagtcag ggccgtcaga cctagggcta
      721 ggtgaatgcc ccatcctgca caccctcctt ccctttccca caaagtctgc agtttgggat
      781 ccgccaccat ggccgatgct aagaacatta agaagggccc tgctcccttc taccctctgg
      841 aggatggcac cgctggcgag cagctgcaca aggccatgaa gaggtatgcc ctggtgcctg
      901 gcaccattgc cttcaccgat gcccacattg aggtggacat cacctatgcc gagtacttcg
      961 agatgtctgt gcgcctggcc gaggccatga agaggtacgg cctgaacacc aaccaccgca
     1021 tcgtggtgtg ctctgagaac tctctgcagt tcttcatgcc agtgctgggc gccctgttca
     1081 tcggagtggc cgtggcccct gctaacgaca tttacaacga gcgcgagctg ctgaacagca
     1141 tgggcatttc tcagcctacc gtggtgttcg tgtctaagaa gggcctgcag aagatcctga
     1201 acgtgcagaa gaagctgcct atcatccaga agatcatcat catggactct aagaccgact
     1261 accagggctt ccagagcatg tacacattcg tgacatctca tctgcctcct ggcttcaacg
     1321 agtacgactt cgtgccagag tctttcgaca gggacaaaac cattgccctg atcatgaaca
     1381 gctctgggtc taccggcctg cctaagggcg tggccctgcc tcatcgcacc gcctgtgtgc
     1441 gcttctctca cgcccgcgac cctattttcg gcaaccagat catccccgac accgctattc
     1501 tgagcgtggt gccattccac cacggcttcg gcatgttcac caccctgggc tacctgattt
     1561 gcggctttcg ggtggtgctg atgtaccgct tcgaggagga gctgttcctg cgcagcctgc
     1621 aagactacaa aattcagtct gccctgctgg tgccaaccct gttcagcttc ttcgctaaga
     1681 gcaccctgat cgacaagtac gacctgtcta acctgcacga gattgcctct ggcggcgccc
     1741 cactgtctaa ggaggtgggc gaagccgtgg ccaagcgctt tcatctgcca ggcatccgcc
     1801 agggctacgg cctgaccgag acaaccagcg ccattctgat taccccagag ggcgacgaca
     1861 agcctggcgc cgtgggcaag gtggtgccat tcttcgaggc caaggtggtg gacctggaca
     1921 ccggcaagac cctgggagtg aaccagcgcg gcgagctgtg tgtgcgcggc cctatgatta
     1981 tgtccggcta cgtgaataac cctgaggcca caaacgccct gatcgacaag gacggctggc
     2041 tgcactctgg cgacattgcc tactgggacg aggacgagca cttcttcatc gtggaccgcc
     2101 tgaagtctct gatcaagtac aagggctacc aggtggcccc agccgagctg gagtctatcc
     2161 tgctgcagca ccctaacatt ttcgacgccg gagtggccgg cctgcccgac gacgatgccg
     2221 gcgagctgcc tgccgccgtc gtcgtgctgg aacacggcaa gaccatgacc gagaaggaga
     2281 tcgtggacta tgtggccagc caggtgacaa ccgccaagaa gctgcgcggc ggagtggtgt
     2341 tcgtggacga ggtgcccaag ggcctgaccg gcaagctgga cgcccgcaag atccgcgaga
     2401 tcctgatcaa ggctaagaaa ggcggcaaga tcgccgtgta actcgaggca ataaaatatc
     2461 ttatttttca ttacatctgt gtgttggttt tttgtgtgtt cttaattaac tgtggaatgt
     2521 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2581 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2641 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2701 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2761 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     2821 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     2881 atctgatcgg cgcgccatac cggtgccacc atggcttcca aggtgtacga ccccgagcaa
     2941 cgcaaacgca tgatcactgg gcctcagtgg tgggctcgct gcaagcaaat gaacgtgctg
     3001 gactccttca tcaactacta tgattccgag aagcacgccg agaacgccgt gatttttctg
     3061 catggtaacg ctgcctccag ctacctgtgg aggcacgtcg tgcctcacat cgagcccgtg
     3121 gctagatgca tcatccctga tctgatcgga atgggtaagt ccggcaagag cgggaatggc
     3181 tcatatcgcc tcctggatca ctacaagtac ctcaccgctt ggttcgagct gctgaacctt
     3241 ccaaagaaaa tcatctttgt gggccacgac tggggggctt gtctggcctt tcactactcc
     3301 tacgagcacc aagacaagat caaggccatc gtccatgctg agagtgtcgt ggacgtgatc
     3361 gagtcctggg acgagtggcc tgacatcgag gaggatatcg ccctgatcaa gagcgaagag
     3421 ggcgagaaaa tggtgcttga gaataacttc ttcgtcgaga ccatgctccc aagcaagatc
     3481 atgcggaaac tggagcctga ggagttcgct gcctacctgg agccattcaa ggagaagggc
     3541 gaggttagac ggcctaccct ctcctggcct cgcgagatcc ctctcgttaa gggaggcaag
     3601 cccgacgtcg tccagattgt ccgcaactac aacgcctacc ttcgggccag cgacgatctg
     3661 cctaagatgt tcatcgagtc cgaccctggg ttcttttcca acgctattgt cgagggagct
     3721 aagaagttcc ctaacaccga gttcgtgaag gtgaagggcc tccacttcag ccaggaggac
     3781 gctccagatg aaatgggtaa gtacatcaag agcttcgtgg agcgcgtgct gaagaacgag
     3841 cagtaagcgg ccgcgatccc gcccctctcc ctcccccccc cctaacgtta ctggccgaag
     3901 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     3961 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     4021 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     4081 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     4141 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     4201 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     4261 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     4321 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     4381 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg
     4441 ccacaaccca caaggagacg accttccatg accgagtaca agcccacggt gcgcctcgcc
     4501 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     4561 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4621 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4681 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4741 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     4801 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4861 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     4921 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4981 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     5041 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
     5101 aggagcgcac gaccccatgc atcggtacct agagtcgggg cggccggccg cttcgagcag
     5161 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     5221 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5281 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     5341 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgctgca gctctggccc
     5401 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     5461 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     5521 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     5581 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     5641 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     5701 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     5761 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     5821 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     5881 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     5941 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     6001 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     6061 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     6121 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     6181 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     6241 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     6301 tttttctaat cagaattggt taattggttg taacattatt cagattgggc cccgttccac
     6361 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6421 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6481 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6541 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6601 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6661 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6721 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6781 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6841 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6901 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6961 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7021 gccttttgct ggccttttgc tcacatgtt
//
