LOCUS       62056_18805        6383 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 6383)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..328
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             425..1021
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    1254..1335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1588..1945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1967..2665
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    complement(2692..2813)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2974..3831
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4005..4593
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4881..4902
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4917..4947
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4955..4971
                     /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)."
     primer_bind     4979..4995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5015..5033
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        5045..5063
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5124..5172
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    5186..5277
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     promoter        5406..5437
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             5476..6192
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 gacgaaagat tggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
       61 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
      121 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
      181 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
      241 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
      301 cttttttgga ggcctaggct tttgcaaaaa gctagcttgc atgcctgcag gtcggccgcc
      361 acgaccggtg ccgccaccat cccctgaccc acgcccctga cccctcacaa ggagacgacc
      421 ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
      481 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
      541 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
      601 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
      661 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
      721 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
      781 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
      841 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
      901 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
      961 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     1021 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
     1081 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac
     1141 tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1201 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1261 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1321 catttttttc actgcaatct cgtgatacgc ctatttttat aggttaatgt catgataata
     1381 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1441 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1501 cttcaataat atctcgtcat gtgggcccat ctgcgcagca ccatggcctg aaataacctc
     1561 tgaaagagga acttggttag ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     1621 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1681 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1741 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     1801 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     1861 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     1921 ttttggaggc ctaggctttt gcaaaaagct cgattcttct gacacgatga gcgagctgat
     1981 taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa
     2041 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     2101 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg
     2161 cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc
     2221 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     2281 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     2341 cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga
     2401 gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt
     2461 gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa
     2521 gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
     2581 caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc
     2641 tagcaaactg gggcacaagc ttaattaagc ggccgcttcg agcagacatg ataagataca
     2701 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2761 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2821 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2881 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcg atccaggtgg cactttgggc
     2941 ccatctgctg tcatatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3001 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3061 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3121 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3181 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3241 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3301 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3361 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3421 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3481 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3541 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3601 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3661 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3721 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3781 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3841 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttttttg
     4081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4141 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4681 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4741 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4801 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4861 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     4921 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     4981 ggaaacagct atgacatgat tacgaattgc aacgatttag gtgacactat agaagagaag
     5041 gaattaatac gactcactat agggagagag agagaattac cctcactaaa gggaggagaa
     5101 gcatgaattc ggcctaactg gccaataaaa tatctttatt ttcattacat ctgtgtgttg
     5161 gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa
     5221 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctggc
     5281 ctaactggcc gtgacactgc agcagtgact acgcatgcgt gacgatgcag acgtgacact
     5341 gcactggtga caaagcaccc gtgactaggc aagcgtgacg tagcatcggt gacacagcaa
     5401 gacactagag ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt
     5461 aaactcgagg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     5521 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     5581 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     5641 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     5701 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     5761 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     5821 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     5881 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     5941 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     6001 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     6061 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     6121 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     6181 gagctgtaca agtaacatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6241 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     6301 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     6361 accgtcatca ccgaaacgcg cga
//
