LOCUS       62056_19045        3561 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 3561)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             169..192
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     CDS             199..885
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTG"
     terminator      976..1023
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1120..1575
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1601..1705
                     /label=AmpR promoter
     CDS             1706..2563
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2737..3325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccgat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
      241 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      301 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      361 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      421 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
      481 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
      541 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
      601 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
      661 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
      721 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
      781 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
      841 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcgcatg gacgagctgt
      901 acaagtaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      961 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1021 ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc
     1081 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     1141 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1201 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1261 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1321 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1381 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1441 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1501 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1561 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1681 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1801 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1861 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1921 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1981 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2101 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2281 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2401 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2581 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2641 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2701 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2761 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2821 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2881 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2941 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3001 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3061 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3121 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3181 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3241 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3301 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3361 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3421 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3481 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3541 ttggccgatt cattaatgca g
//
