LOCUS       62056_6510        5862 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 5862)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             969..1514
                     /codon_start=1
                     /label=CLIP-tag(TM)
                     /note="human O6-alkylguanine-DNA-alkyltransferase variant
                     that forms covalent bonds with benzylcytosine derivatives"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          1638..1867
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     misc_feature    1945..2526
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2549..3349
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3482..3603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3705..3721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3729..3745)
                     /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(3753..3783)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3798..3819)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4107..4695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4869..5726)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5727..5831)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgtcgctagc gatatcggcg cgccagcatt taaatctgta cagaccggtg
      961 aattcaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
     1021 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
     1081 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
     1141 cactgatcca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
     1201 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
     1261 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagcgag agccacctgg
     1321 ccgccctggt gggcaatccc gccgccaccg ccgccgtgaa caccgccctg gacggaaatc
     1381 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacagcgac gtggggccct
     1441 acctgggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
     1501 agcctgggct gggtcctgca ggcggatccg cgtttaaact cgaggttaat taatgagcgg
     1561 ccgcttaagc cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc
     1621 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag
     1681 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt
     1741 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg
     1801 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc
     1861 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat
     1921 tccgcccctc tccccccccc ccttttccct cccccccccc taacgttact ggccgaagcc
     1981 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     2041 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     2101 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     2161 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     2221 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     2281 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2341 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     2401 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     2461 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc
     2521 acaacccggg ataattcctg cagccaatat gggatcggcc attgaacaag atggattgca
     2581 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2641 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2701 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     2761 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     2821 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     2881 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     2941 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3001 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3061 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca
     3121 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3181 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3241 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3301 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatca
     3361 attctctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3421 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3481 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3601 ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3661 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3721 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3781 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3841 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3901 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3961 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4021 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4081 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4141 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4201 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4261 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4321 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4381 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4441 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4501 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4561 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4621 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4681 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4741 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4801 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
