LOCUS       62056_22295        5216 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 5216)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(435..512)
                     /label=lacI promoter
     promoter        558..586
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             633..1340
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1371..2087
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(2118..2819)
                     /codon_start=1
                     /label=TagBFP2
                     /note="Also known as mTagBFP2; improved variant of TagBFP 
                     generated by I174A mutation"
                     /translation="SIKLVPQFAREVAVSGHCHLVLLDVGLVVGLLDSFQSVVHIVDAR
                     HLEVLSGFLGSICGLGVCDQMAPAHELQGHVVSAFQAAVSGVQRLGEGLPAECFLLHHR
                     AVGCEVHPSDLDVVDEAAVLEAGVLGSGQHAPVFVCGDSLPCEALREGLLEEVGDALGV
                     VDEGLAAVEEASSQDVEGEGERAALDHLDSHGLGALVGLAFALGCALEVMVVHGALHVQ
                     LHVHVLLNQLAH"
     terminator      2853..2900
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2995..3011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3019..3035)
                     /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(3043..3073)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3088..3109)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3397..3985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4159..5016)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5017..5121)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
      481 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
      541 ctcctgcatt aggaaatttg acaattaatc atcggctcgt ataatgtgtg gaatgtctca
      601 tgagcggata tttcacacag gaaacagtat tcatggtgag caagggcgag gaggataaca
      661 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
      721 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      781 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
      841 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
      901 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
      961 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     1021 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1081 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     1141 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     1201 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     1261 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     1321 gcatggacga gctgtacaag ggcggtggag gaagcggcgg aggtggatct atggtgagca
     1381 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1441 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1501 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1561 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1621 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1681 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1741 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1801 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1861 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1921 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1981 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2041 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtggaggct
     2101 ctggtggcgg tggatcaatg agcgagctga ttaaggagaa catgcacatg aagctgtaca
     2161 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
     2221 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     2281 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
     2341 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     2401 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     2461 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
     2521 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg
     2581 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca
     2641 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
     2701 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg
     2761 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattgat
     2821 taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     2881 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacacatc
     2941 ggatcccggg cccgtcgact gcagaggcct gcatgcaagc ttggcgtaat catggtcata
     3001 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3061 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3121 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3181 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3241 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3301 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3361 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3421 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3481 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3541 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3601 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3661 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3721 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3781 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3841 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3901 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3961 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4021 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4081 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4141 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4201 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4261 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4321 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4381 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4441 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4501 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4561 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4621 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4681 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4741 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4801 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4861 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4921 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4981 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5041 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5101 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5161 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
