LOCUS       62056_16615        6962 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 6962)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6962
                     /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             702..2351
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        2432..2761
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2824..3756
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    3775..4361
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4381..4977
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    complement(5080..5201)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5413..6219
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6312..6900
                     /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 tatttaaatg tcgacgttta aactctagag gatccgccac catggccgat gctaagaaca
      721 ttaagaaggg ccctgctccc ttctaccctc tggaggatgg caccgctggc gagcagctgc
      781 acaaggccat gaagaggtat gccctggtgc ctggcaccat tgccttcacc gatgcccaca
      841 ttgaggtgga catcacctat gccgagtact tcgagatgtc tgtgcgcctg gccgaggcca
      901 tgaagaggta cggcctgaac accaaccacc gcatcgtggt gtgctctgag aactctctgc
      961 agttcttcat gccagtgctg ggcgccctgt tcatcggagt ggccgtggcc cctgctaacg
     1021 acatttacaa cgagcgcgag ctgctgaaca gcatgggcat ttctcagcct accgtggtgt
     1081 tcgtgtctaa gaagggcctg cagaagatcc tgaacgtgca gaagaagctg cctatcatcc
     1141 agaagatcat catcatggac tctaagaccg actaccaggg cttccagagc atgtacacat
     1201 tcgtgacatc tcatctgcct cctggcttca acgagtacga cttcgtgcca gagtctttcg
     1261 acagggacaa aaccattgcc ctgatcatga acagctctgg gtctaccggc ctgcctaagg
     1321 gcgtggccct gcctcatcgc accgcctgtg tgcgcttctc tcacgcccgc gaccctattt
     1381 tcggcaacca gatcatcccc gacaccgcta ttctgagcgt ggtgccattc caccacggct
     1441 tcggcatgtt caccaccctg ggctacctga tttgcggctt tcgggtggtg ctgatgtacc
     1501 gcttcgagga ggagctgttc ctgcgcagcc tgcaagacta caaaattcag tctgccctgc
     1561 tggtgccaac cctgttcagc ttcttcgcta agagcaccct gatcgacaag tacgacctgt
     1621 ctaacctgca cgagattgcc tctggcggcg ccccactgtc taaggaggtg ggcgaagccg
     1681 tggccaagcg ctttcatctg ccaggcatcc gccagggcta cggcctgacc gagacaacca
     1741 gcgccattct gattacccca gagggcgacg acaagcctgg cgccgtgggc aaggtggtgc
     1801 cattcttcga ggccaaggtg gtggacctgg acaccggcaa gaccctggga gtgaaccagc
     1861 gcggcgagct gtgtgtgcgc ggccctatga ttatgtccgg ctacgtgaat aaccctgagg
     1921 ccacaaacgc cctgatcgac aaggacggct ggctgcactc tggcgacatt gcctactggg
     1981 acgaggacga gcacttcttc atcgtggacc gcctgaagtc tctgatcaag tacaagggct
     2041 accaggtggc cccagccgag ctggagtcta tcctgctgca gcaccctaac attttcgacg
     2101 ccggagtggc cggcctgccc gacgacgatg ccggcgagct gcctgccgcc gtcgtcgtgc
     2161 tggaacacgg caagaccatg accgagaagg agatcgtgga ctatgtggcc agccaggtga
     2221 caaccgccaa gaagctgcgc ggcggagtgg tgttcgtgga cgaggtgccc aagggcctga
     2281 ccggcaagct ggacgcccgc aagatccgcg agatcctgat caaggctaag aaaggcggca
     2341 agatcgccgt gtaactcgag gcaataaaat atcttatttt tcattacatc tgtgtgttgg
     2401 ttttttgtgt gttcttaatt aactgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2461 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2521 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2581 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2641 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2701 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2761 aaagctcccg ggagcttgta tatccatttt cggatctgat cggcgcgcca taccggtgcc
     2821 accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac tgggcctcag
     2881 tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta ctatgattcc
     2941 gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc cagctacctg
     3001 tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc tgatctgatc
     3061 ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga tcactacaag
     3121 tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt tgtgggccac
     3181 gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa gatcaaggcc
     3241 atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg gcctgacatc
     3301 gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct tgagaataac
     3361 ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc tgaggagttc
     3421 gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac cctctcctgg
     3481 cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat tgtccgcaac
     3541 tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga gtccgaccct
     3601 gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac cgagttcgtg
     3661 aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg taagtacatc
     3721 aagagcttcg tggagcgcgt gctgaagaac gagcagtaag cggccgcgat cccgcccctc
     3781 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     3841 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     3901 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     3961 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4021 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4081 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4141 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     4201 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     4261 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     4321 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccacaaggag acgaccttcc
     4381 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     4441 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     4501 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4561 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4621 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4681 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     4741 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4801 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     4861 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4921 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     4981 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcggta
     5041 cctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     5101 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     5161 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     5221 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     5281 acaaatgtgg taaaatcgct gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     5341 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     5401 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     5461 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     5521 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     5581 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     5641 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     5701 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     5761 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     5821 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     5881 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     5941 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     6001 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     6061 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     6121 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     6181 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     6241 ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt agaaaagatc
     6301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6421 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6661 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6961 tt
//
