LOCUS       62056_19030        5761 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 5761)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5761
                     /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      3176..3223
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3320..3775
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3801..3905
                     /label=AmpR promoter
     CDS             3906..4763
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4937..5525
                     /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 gatgggcccg gggggaccct gtaccccggt ggggtggccg
      961 ctgcctcttc tgctggtgat ggcggctggg gtggctccgg tgtgggcctc tcactcccct
     1021 catctcccgc ggcctcaccc gagggtcccc ccgcacccct cctcagaacg gcgtgcagta
     1081 tacatcgggg cgctgtttcc catgagcggg ggctggccgg ggggccaggc ctgccagcct
     1141 gcggtggaga tggcgctgga ggacgttaac agccgcagag acatcctgcc ggactacgag
     1201 ctcaagctta tccaccacga cagcaagtgc gacccagggc aagccaccaa gtacttgtat
     1261 gaactactct acaacgaccc catcaagatc atcctcatgc ccggctgcag ctctgtgtcc
     1321 acactggtag ccgaggctgc ccggatgtgg aaccttattg tgctctcata tggctccagc
     1381 tcaccagcct tgtcaaaccg acagcggttt ccaacgttct ttcggacaca tccatccgcc
     1441 acactccaca atcccacccg ggtgaaactc ttcgaaaagt ggggctggaa gaagattgcc
     1501 accatccagc agactaccga ggtcttcacc tcaacactgg atgacctgga ggagcgagtg
     1561 aaagaggctg ggattgagat cacttttcga cagagtttct tctcagatcc agctgtgcct
     1621 gttaaaaacc tgaagcgtca agatgctcga atcatcgtgg gacttttcta tgagaccgaa
     1681 gcccggaaag ttttttgtga ggtctataag gaacggctct ttgggaagaa gtatgtctgg
     1741 tttctcatcg ggtggtatgc tgacaactgg ttcaaaacct atgacccgtc aatcaattgt
     1801 acagtagaag agatgactga ggcggtggag ggccatatca ccacggagat tgtcatgctg
     1861 aaccctgcca acacccgaag catttccaac atgacatcac aggaatttgt ggagaaacta
     1921 accaagcggc tgaaaagaca ccctgaggag actggaggct tccaggaggc accactggcc
     1981 tatgatgcta tctgggcctt ggctttggcc ttgaacaaga cctctggagg aggtggccgt
     2041 tcaggagtgc gcctggagga ctttaactac aacaaccaga ccattacaga ccaaatctac
     2101 cgggccatga actcctcctc ctttgagggt gtttctggcc acgtggtctt tgatgccagc
     2161 ggctcccgga tggcatggac gcttatcgag cagctacagg gcggcagcta caagaagatc
     2221 ggctactacg acagcaccaa ggatgatctt tcctggtcca aaacagacaa gtggatcgga
     2281 gggtctcccc cagccgacca gaccttggtc atcaagacat tccgtttcct gtcacagaaa
     2341 ctctttatct ccgtctcagt tctctccagc ctgggcggta ccatgtctaa aggtgaagaa
     2401 ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
     2461 ttttctgtct ccggtgaagg tgaaggtgat gctacgtacg gtaaattgac cttaaaatta
     2521 ctctgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat
     2581 ggtgttcaat gttttgctag atacccagat catatgaaac aacatgactt tttcaagtct
     2641 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
     2701 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
     2761 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac
     2821 tctcacaatg tttacatcac tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa
     2881 attagacaca acattgaaga tggtggtgtt caattagctg accattatca acaaaatact
     2941 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccta tcaatctgcc
     3001 ttattcaaag atccaaacga aaagagagac cacatggtct tgttagaatt tttgactgct
     3061 gctggtatta ccgagggtat gaatgaattg tacaaatgag aattcgaagc ttgatccggc
     3121 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     3181 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     3241 atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3301 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3361 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3421 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3481 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3541 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3601 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3661 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3721 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3781 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3841 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3901 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3961 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4021 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4081 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4141 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4201 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4261 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4321 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4381 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4441 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4501 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4561 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4621 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4681 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4741 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4801 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4861 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4921 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5161 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5221 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5281 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5341 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5401 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5461 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5521 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5581 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5641 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5701 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5761 g
//
