LOCUS       62056_19025        5641 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 5641)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5641
                     /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..915
                     /codon_start=1
                     /label=ECFP
                     /note="enhanced CFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      3056..3103
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3200..3655
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3681..3785
                     /label=AmpR promoter
     CDS             3786..4643
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4817..5405
                     /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 ggcgaggagc tgttcaccgg ggtggtgccc
      241 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      301 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      361 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc
      421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      481 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      601 ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc
      661 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
      721 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      781 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
      841 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      901 gacgagctgt acaagctcga gtctcactcc cctcatctcc cgcggcctca cccgagggtc
      961 cccccgcacc cctcctcaga acggcgtgca gtatacatcg gggcgctgtt tcccatgagc
     1021 gggggctggc cggggggcca ggcctgccag cctgcggtgg agatggcgct ggaggacgtt
     1081 aacagccgca gagacatcct gccggactac gagctcaagc ttatccacca cgacagcaag
     1141 tgcgacccag ggcaagccac caagtacttg tatgaactac tctacaacga ccccatcaag
     1201 atcatcctca tgcccggctg cagctctgtg tccacactgg tagccgaggc tgcccggatg
     1261 tggaacctta ttgtgctctc atatggctcc agctcaccag ccttgtcaaa ccgacagcgg
     1321 tttccaacgt tctttcggac acatccatcc gccacactcc acaatcccac ccgggtgaaa
     1381 ctcttcgaaa agtggggctg gaagaagatt gccaccatcc agcagactac cgaggtcttc
     1441 acctcaacac tggatgacct ggaggagcga gtgaaagagg ctgggattga gatcactttt
     1501 cgacagagtt tcttctcaga tccagctgtg cctgttaaaa acctgaagcg tcaagatgct
     1561 cgaatcatcg tgggactttt ctatgagacc gaagcccgga aagttttttg tgaggtctat
     1621 aaggaacggc tctttgggaa gaagtatgtc tggtttctca tcgggtggta tgctgacaac
     1681 tggttcaaaa cctatgaccc gtcaatcaat tgtacagtag aagagatgac tgaggcggtg
     1741 gagggccata tcaccacgga gattgtcatg ctgaaccctg ccaacacccg aagcatttcc
     1801 aacatgacat cacaggaatt tgtggagaaa ctaaccaagc ggctgaaaag acaccctgag
     1861 gagactggag gcttccagga ggcaccactg gcctatgatg ctatctgggc cttggctttg
     1921 gccttgaaca agacctctgg aggaggtggc cgttcaggag tgcgcctgga ggactttaac
     1981 tacaacaacc agaccattac agaccaaatc taccgggcca tgaactcctc ctcctttgag
     2041 ggtgtttctg gccacgtggt ctttgatgcc agcggctccc ggatggcatg gacgcttatc
     2101 gagcagctac agggcggcag ctacaagaag atcggctact acgacagcac caaggatgat
     2161 ctttcctggt ccaaaacaga caagtggatc ggagggtctc ccccagccga ccagaccttg
     2221 gtcatcaaga cattccgttt cctgtcacag aaactcggta ccatgtctaa aggtgaagaa
     2281 ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
     2341 ttttctgtct ccggtgaagg tgaaggtgat gctacgtacg gtaaattgac cttaaaatta
     2401 ctctgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat
     2461 ggtgttcaat gttttgctag atacccagat catatgaaac aacatgactt tttcaagtct
     2521 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
     2581 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
     2641 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac
     2701 tctcacaatg tttacatcac tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa
     2761 attagacaca acattgaaga tggtggtgtt caattagctg accattatca acaaaatact
     2821 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccta tcaatctgcc
     2881 ttattcaaag atccaaacga aaagagagac cacatggtct tgttagaatt tttgactgct
     2941 gctggtatta ccgagggtat gaatgaattg tacaaatgag aattcgaagc ttgatccggc
     3001 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     3061 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     3121 atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3181 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3241 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3301 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3361 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3421 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3481 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3541 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3601 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5521 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5581 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5641 g
//
