LOCUS       62056_6820        6084 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 6084)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(275..302)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(394..480)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     544..560
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    576..675
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             742..1452
                     /codon_start=1
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSDGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1510..2106
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     protein_bind    complement(2120..2219)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2265..2644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2645..2848
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2893..2911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3010..3726
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3738..4325
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        complement(4333..4351)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4356..4372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4485..5291
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5441..6029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
       61 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      121 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      181 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
      241 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
      301 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
      361 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
      421 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
      481 atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac
      541 gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga
      601 tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt
      661 acaaaaaagc aggctggatc cggtaccggc cggcttgtcg acgacggcgg tctccgtcgt
      721 caggatcatc cggccgccac catggtgtct aagggcgaag agctgattaa ggagaacatg
      781 cacatgaagc tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgat
      841 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
      901 cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc
      961 atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccctga gggcttcaca
     1021 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     1081 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac
     1141 ggccctgtga tgcagaagaa aacactcggc tgggaggcct ccaccgagat gctgtacccc
     1201 gctgacggcg gcctggaagg cagaagcgac atggccctga agctcgtggg cgggggccac
     1261 ctgatctgca acttgaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg
     1321 cccggcgtct actatgtgga cagaagactg gaaagaatca aggaggccga caaagagacc
     1381 tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg
     1441 cacaaactta attccgaggg ccgcggcagc ctgctgacct gcggcgacgt ggaggaaaac
     1501 cccggcccca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     1561 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     1621 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     1681 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     1741 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     1801 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     1861 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     1921 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     1981 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     2041 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     2101 ggtgcctgag aattctacca cccagctttc ttgtacaaag ttggcattat aagaaagcat
     2161 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     2221 catccagctg atggggcaca aacttaattc ctgataagcg cgttgacatt gattattgac
     2281 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     2341 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2401 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2461 atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     2521 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     2581 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     2641 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     2701 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     2761 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     2821 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     2881 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt
     2941 aagcttggta ccgaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3001 aaatctagaa tggtgagcaa gggcgaggag ctgttcaccg gtgtggtgcc catcctggtc
     3061 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3121 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3181 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3241 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3301 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3361 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3421 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3481 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3541 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3601 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3661 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3721 tacaagtaag cggccgcaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3781 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3841 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3901 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3961 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4021 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4081 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     4141 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4201 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4261 ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg gatctccctt tggccgcctc
     4321 cccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     4381 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     4441 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     4501 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     4561 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     4621 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     4681 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     4741 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     4801 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     4861 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     4921 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     4981 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     5041 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     5101 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     5161 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     5221 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     5281 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     5341 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     5401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5581 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6061 tggccttttg ctggcctttt gctc
//
