LOCUS       62056_11710        7578 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 7578)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..323
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        324..527
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             557..1273
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1399..1520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1645..1676
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             1709..3358
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             3362..3481
                     /codon_start=1
                     /label=hPEST
                     /note="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    complement(3533..3654)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3848..4205
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4236..5270
                     /codon_start=1
                     /label=HygR
                     /note="hygromycin B phosphotransferase"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     polyA_signal    5297..5345
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5672..6260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6463..7320)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    7487..7578
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 ggcctaactg gccggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
       61 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      361 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      421 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      481 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      541 gtcagatccc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      601 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      661 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      721 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      781 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      841 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      901 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      961 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1021 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1081 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1141 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1201 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1261 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1321 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1381 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1441 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1501 aaactcatca atgtatctta gctagcgcac cagacagtga cgtcagctgc cagatcccat
     1561 ggccgtcata ctgtgacgtc tttcagacac cccattgacg tcaatgggag aacagatctg
     1621 gcctcggcgg ccaagcttag acactagagg gtatataatg gaagctcgac ttccagcttg
     1681 gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta agaagggccc
     1741 agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa
     1801 gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat
     1861 tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg
     1921 gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc
     1981 cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga
     2041 gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa
     2101 agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat
     2161 catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca
     2221 tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac
     2281 catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc
     2341 gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat
     2401 catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac
     2461 cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga
     2521 gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact
     2581 atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga
     2641 gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt
     2701 ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat
     2761 cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc
     2821 taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg
     2881 cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct
     2941 catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca
     3001 cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc
     3061 agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg
     3121 cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa
     3181 aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa
     3241 gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga
     3301 cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgaa
     3361 ttctcacggc ttccctcccg aggtggagga gcaggccgcc ggcaccctgc ccatgagctg
     3421 cgcccaggag agcggcatgg atagacaccc tgctgcttgc gccagcgcca ggatcaacgt
     3481 ctaaggccgc gactctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     3541 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3601 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3661 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3721 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc
     3781 ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     3841 ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3961 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4021 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4081 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4141 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4201 gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta
     4261 ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt
     4321 cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc
     4381 gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca
     4441 gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat
     4501 actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc
     4561 ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc
     4621 aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg
     4681 atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca
     4741 ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc
     4801 ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg
     4861 gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg
     4921 tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact
     4981 tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca
     5041 tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg
     5101 ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa
     5161 tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg
     5221 gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag
     5281 aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     5341 gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     5401 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     5461 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     5521 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6421 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     6481 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     6541 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     6601 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     6661 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     6721 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     6781 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     6841 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     6901 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     6961 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     7021 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     7081 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     7141 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     7201 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     7261 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     7321 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
     7381 ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa atatctttat
     7441 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     7501 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     7561 gtgccagaac atttctct
//
