LOCUS       62056_2245        4648 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 4648)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     508..524
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        621..1292
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     CDS             1449..1475
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     CDS             1500..2216
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2235..2252
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     primer_bind     complement(2556..2572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2580..2596)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2604..2634)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2649..2670)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2958..3546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3720..4577)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(4578..4648,1..34))
                     /label=AmpR promoter
ORIGIN
        1 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
       61 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      121 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      181 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      241 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      301 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      361 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      421 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      481 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgcatgcc
      541 tgcagtcaca tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc
      601 tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg aaacctcctc
      661 ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc
      721 tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac
      781 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
      841 accacgtctt caaagcaagt ggattgatgt gataacatgg tggagcacga cacacttgtc
      901 tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga gacttttcaa
      961 caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg tcactttatt
     1021 gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
     1081 gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg
     1141 agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat
     1201 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
     1261 atataaggaa gttcatttca tttggagagg acctcgacct caacacaaca tatacaaaac
     1321 aaacgaatct caagcaatca agcattctac ttctattgca gcaatttaaa tcatttcttt
     1381 taaagcaaaa gcaattttct gaaaattttc accatttacg aacgatactc gagatgaccc
     1441 gtggttctca tcaccatcac catcaccatc accatcacgc catggtcgac gtactagtga
     1501 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1561 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1621 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1681 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1741 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1801 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1861 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1921 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1981 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2041 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2101 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2161 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtc
     2221 tagacccggg aatgcatcac catcaccatc acggatcctg attgatcgat agagctcgaa
     2281 tttccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     2341 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     2401 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     2461 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     2521 gtcatctatg tacagataga gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2581 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2761 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3361 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4621 tgtctcatga gcggatacat atttgaat
//
