LOCUS       62056_19040        6031 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 6031)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6031
                     /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      3446..3493
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3590..4045
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4071..4175
                     /label=AmpR promoter
     CDS             4176..5033
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5207..5795
                     /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 gggcggggcg cagaccccca acgccacctc ggaaggttgc
      961 cagattatac atccgccctg ggaaggtggc atcaggtacc gtggcttgac tcgcgaccag
     1021 gtgaaggcca tcaacttcct gcctgtggac tatgagatcg aatatgtgtg ccgaggggag
     1081 cgcgaggtgg tggggcccaa ggtgcgcaaa tgcctggcca acggctcctg gacggatatg
     1141 gacacaccca gccgctgtgt ccgaatctgc tccaagtctt atttgaccct ggaaaatggg
     1201 aaggttttcc tgacgggtgg ggacctccca gctctggatg gagcccgggt ggagttccga
     1261 tgtgaccccg acttccatct ggtgggcagc tcccggagcg tctgtagtca gggccagtgg
     1321 agcaccccca agccccactg ccaggtgaat cgaacgccac actcagaacg gcgtgcagta
     1381 tacatcgggg cgctgtttcc catgagcggg ggctggccgg ggggccaggc ctgccagccc
     1441 gcggtggaga tggcgctgga ggacgttaac agccgcagag acatcctgcc ggactacgag
     1501 ctcaagctta tccaccacga cagcaagtgt gacccagggc aagccaccaa gtacttgtac
     1561 gaactactct acaatgaccc catcaagatc attctcatgc ctggctgtag ttctgtctcc
     1621 acacttgtag ctgaggctgc ccggatgtgg aaccttattg tgctctcata tggctccagt
     1681 tcaccagcct tgtcaaaccg acagcggttt cccacgttct tccggacgca tccatccgcc
     1741 acactccaca atcccacccg ggtgaaactc ttcgaaaagt ggggctggaa gaagatcgct
     1801 accatccaac agaccaccga ggtcttcacc tcaacgctgg atgacctgga ggagcgagtg
     1861 aaagaggctg ggatcgagat cactttccga cagagtttct tctcggatcc agctgtgcct
     1921 gttaaaaacc tgaagcgtca agatgctcga atcatcgtgg gacttttcta tgagacggaa
     1981 gcccggaaag ttttttgtga ggtctataag gaaaggctct ttgggaagaa gtacgtctgg
     2041 ttcctcatcg ggtggtatgc tgacaactgg ttcaagacct atgacccgtc aatcaattgt
     2101 acagtggaag aaatgaccga ggcggtggag ggccacatca ccacggagat tgtcatgctg
     2161 aaccctgcca acacccgaag catttccaac atgacgtcac aggaatttgt ggagaaacta
     2221 accaagcggc tgaaaagaca ccccgaggag actggaggct tccaggaggc accactggcc
     2281 tatgatgcta tctgggcctt ggctttggcc ttgaacaaga cgtctggagg aggtggtcgt
     2341 tccggcgtgc gcctggagga ctttaactac aacaaccaga ccattacaga ccagatctac
     2401 cgggccatga actcctcctc ctttgagggc gtttctggcc atgtggtctt tgatgccagc
     2461 ggctcccgga tggcatggac acttatcgag cagctacagg gcggcagcta caagaagatc
     2521 ggctactacg acagcaccaa ggatgatctt tcctggtcca aaacggacaa gtggattgga
     2581 gggtctcccc cagctgacca gaccttggtc atcaagacat tccgtttcct gtctcagaaa
     2641 ctctttggta ccatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt
     2701 gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat
     2761 gctacgtacg gtaaattgac cttaaaatta ctctgtacta ctggtaaatt gccagttcca
     2821 tggccaacct tagtcactac tttaggttat ggtgttcaat gttttgctag atacccagat
     2881 catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga
     2941 actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt
     3001 gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt
     3061 ttaggtcaca aattggaata caactataac tctcacaatg tttacatcac tgctgacaaa
     3121 caaaagaatg gtatcaaagc taacttcaaa attagacaca acattgaaga tggtggtgtt
     3181 caattagctg accattatca acaaaatact ccaattggtg atggtccagt cttgttacca
     3241 gacaaccatt acttatccta tcaatctgcc ttattcaaag atccaaacga aaagagagac
     3301 cacatggtct tgttagaatt tttgactgct gctggtatta ccgagggtat gaatgaattg
     3361 tacaaatgag aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3421 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3481 gaggggtttt ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc
     3541 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3601 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3661 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3721 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3781 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3841 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3901 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3961 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4021 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4141 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4201 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4261 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4321 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4381 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4441 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4501 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4561 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4621 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4681 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4741 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4861 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5041 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5341 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5701 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5761 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5821 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5881 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5941 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6001 tccccgcgcg ttggccgatt cattaatgca g
//
