LOCUS       62056_20275        6314 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 6314)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             724..1656
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1657..1680
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     polyA_signal    1729..1777
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        1786..2537
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2573..4222
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    complement(4269..4390)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4523..4627
                     /label=AmpR promoter
     CDS             4628..5485
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5659..6247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcg agcgcagaag caggcagatg agagcaggcc
      721 accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac tgggcctcag
      781 tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta ctatgattcc
      841 gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc cagctacctg
      901 tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc tgatctgatc
      961 ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga tcactacaag
     1021 tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt tgtgggccac
     1081 gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa gatcaaggcc
     1141 atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg gcctgacatc
     1201 gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct tgagaataac
     1261 ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc tgaggagttc
     1321 gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac cctctcctgg
     1381 cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat tgtccgcaac
     1441 tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga gtccgaccct
     1501 gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac cgagttcgtg
     1561 aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg taagtacatc
     1621 aagagcttcg tggagcgcgt gctgaagaac gagcaggatt acaaggacga cgatgacaag
     1681 taactcgagc ccgggaattc gtttaaacct agagcggccg ctggccgcaa taaaatatct
     1741 ttattttcat tacatctgtg tgttggtttt ttgtgtgagg atctaaatga gtcttcggac
     1801 ctcgcggggg ccgcttaagc ggtggttagg gtttgtctga cgcgggggga gggggaagga
     1861 acgaaacact ctcattcgga ggcggctcgg ggtttggtct tggtggccac gggcacgcag
     1921 aagagcgccg cgatcctctt aagcaccccc ccgccctccg tggaggcggg ggtttggtcg
     1981 gcgggtggta actggcgggc cgctgactcg ggcgggtcgc gcgccccaga gtgtgacctt
     2041 ttcggtctgc tcgcagaccc ccgggcggcg ccgccgcggc ggcgacgggc tcgctgggtc
     2101 ctaggctcca tggggaccgt atacgtggac aggctctgga gcatccgcac gactgcggtg
     2161 atattaccgg agaccttctg cgggacgagc cgggtcacgc ggctgacgcg gagcgtccgt
     2221 tgggcgacaa acaccaggac ggggcacagg tacactatct tgtcacccgg aggcgcgagg
     2281 gactgcagga gcttcaggga gtggcgcagc tgcttcatcc ccgtggcccg ttgctcgcgt
     2341 ttgctggcgg tgtccccgga agaaatatat ttgcatgtct ttagttctat gatgacacaa
     2401 accccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc ggggcggcgc
     2461 ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc
     2521 tgcagcgacc cgcttaaaag cttggcattc cggtactgtt ggtaaagcca ccatggccga
     2581 tgctaagaac attaagaagg gccctgctcc cttctaccct ctggaggatg gcaccgctgg
     2641 cgagcagctg cacaaggcca tgaagaggta tgccctggtg cctggcacca ttgccttcac
     2701 cgatgcccac attgaggtgg acatcaccta tgccgagtac ttcgagatgt ctgtgcgcct
     2761 ggccgaggcc atgaagaggt acggcctgaa caccaaccac cgcatcgtgg tgtgctctga
     2821 gaactctctg cagttcttca tgccagtgct gggcgccctg ttcatcggag tggccgtggc
     2881 ccctgctaac gacatttaca acgagcgcga gctgctgaac agcatgggca tttctcagcc
     2941 taccgtggtg ttcgtgtcta agaagggcct gcagaagatc ctgaacgtgc agaagaagct
     3001 gcctatcatc cagaagatca tcatcatgga ctctaagacc gactaccagg gcttccagag
     3061 catgtacaca ttcgtgacat ctcatctgcc tcctggcttc aacgagtacg acttcgtgcc
     3121 agagtctttc gacagggaca aaaccattgc cctgatcatg aacagctctg ggtctaccgg
     3181 cctgcctaag ggcgtggccc tgcctcatcg caccgcctgt gtgcgcttct ctcacgcccg
     3241 cgaccctatt ttcggcaacc agatcatccc cgacaccgct attctgagcg tggtgccatt
     3301 ccaccacggc ttcggcatgt tcaccaccct gggctacctg atttgcggct ttcgggtggt
     3361 gctgatgtac cgcttcgagg aggagctgtt cctgcgcagc ctgcaagact acaaaattca
     3421 gtctgccctg ctggtgccaa ccctgttcag cttcttcgct aagagcaccc tgatcgacaa
     3481 gtacgacctg tctaacctgc acgagattgc ctctggcggc gccccactgt ctaaggaggt
     3541 gggcgaagcc gtggccaagc gctttcatct gccaggcatc cgccagggct acggcctgac
     3601 cgagacaacc agcgccattc tgattacccc agagggcgac gacaagcctg gcgccgtggg
     3661 caaggtggtg ccattcttcg aggccaaggt ggtggacctg gacaccggca agaccctggg
     3721 agtgaaccag cgcggcgagc tgtgtgtgcg cggccctatg attatgtccg gctacgtgaa
     3781 taaccctgag gccacaaacg ccctgatcga caaggacggc tggctgcact ctggcgacat
     3841 tgcctactgg gacgaggacg agcacttctt catcgtggac cgcctgaagt ctctgatcaa
     3901 gtacaagggc taccaggtgg ccccagccga gctggagtct atcctgctgc agcaccctaa
     3961 cattttcgac gccggagtgg ccggcctgcc cgacgacgat gccggcgagc tgcctgccgc
     4021 cgtcgtcgtg ctggaacacg gcaagaccat gaccgagaag gagatcgtgg actatgtggc
     4081 cagccaggtg acaaccgcca agaagctgcg cggcggagtg gtgttcgtgg acgaggtgcc
     4141 caagggcctg accggcaagc tggacgcccg caagatccgc gagatcctga tcaaggctaa
     4201 gaaaggcggc aagatcgccg tgtaataatt ctagagtcgg ggcggccggc cgcttcgagc
     4261 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4321 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4381 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4441 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccagg
     4501 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4561 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4621 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4681 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4741 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4801 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4861 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4921 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4981 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5041 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5101 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5161 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5221 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5281 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5341 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5401 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5461 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5521 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5581 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5641 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5761 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6241 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6301 tcacatggct cgac
//
