LOCUS       62056_20290        6313 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 6313)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6313
                     /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             694..1626
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1627..1650
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     polyA_signal    1728..1776
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        1785..2536
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             2572..4221
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    complement(4268..4389)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4522..4626
                     /label=AmpR promoter
     CDS             4627..5484
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5658..6246
                     /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 taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcaggatt acaaggacga cgatgacaag taagaggaga gcatcctgta tgaggaggag
     1681 agctcgagcc cgggaattcg tttaaaccta gagcggccgc tggccgcaat aaaatatctt
     1741 tattttcatt acatctgtgt gttggttttt tgtgtgagga tctaaatgag tcttcggacc
     1801 tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac gcggggggag ggggaaggaa
     1861 cgaaacactc tcattcggag gcggctcggg gtttggtctt ggtggccacg ggcacgcaga
     1921 agagcgccgc gatcctctta agcacccccc cgccctccgt ggaggcgggg gtttggtcgg
     1981 cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg cgccccagag tgtgaccttt
     2041 tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg gcgacgggct cgctgggtcc
     2101 taggctccat ggggaccgta tacgtggaca ggctctggag catccgcacg actgcggtga
     2161 tattaccgga gaccttctgc gggacgagcc gggtcacgcg gctgacgcgg agcgtccgtt
     2221 gggcgacaaa caccaggacg gggcacaggt acactatctt gtcacccgga ggcgcgaggg
     2281 actgcaggag cttcagggag tggcgcagct gcttcatccc cgtggcccgt tgctcgcgtt
     2341 tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa
     2401 ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg
     2461 gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct
     2521 gcagcgaccc gcttaaaagc ttggcattcc ggtactgttg gtaaagccac catggccgat
     2581 gctaagaaca ttaagaaggg ccctgctccc ttctaccctc tggaggatgg caccgctggc
     2641 gagcagctgc acaaggccat gaagaggtat gccctggtgc ctggcaccat tgccttcacc
     2701 gatgcccaca ttgaggtgga catcacctat gccgagtact tcgagatgtc tgtgcgcctg
     2761 gccgaggcca tgaagaggta cggcctgaac accaaccacc gcatcgtggt gtgctctgag
     2821 aactctctgc agttcttcat gccagtgctg ggcgccctgt tcatcggagt ggccgtggcc
     2881 cctgctaacg acatttacaa cgagcgcgag ctgctgaaca gcatgggcat ttctcagcct
     2941 accgtggtgt tcgtgtctaa gaagggcctg cagaagatcc tgaacgtgca gaagaagctg
     3001 cctatcatcc agaagatcat catcatggac tctaagaccg actaccaggg cttccagagc
     3061 atgtacacat tcgtgacatc tcatctgcct cctggcttca acgagtacga cttcgtgcca
     3121 gagtctttcg acagggacaa aaccattgcc ctgatcatga acagctctgg gtctaccggc
     3181 ctgcctaagg gcgtggccct gcctcatcgc accgcctgtg tgcgcttctc tcacgcccgc
     3241 gaccctattt tcggcaacca gatcatcccc gacaccgcta ttctgagcgt ggtgccattc
     3301 caccacggct tcggcatgtt caccaccctg ggctacctga tttgcggctt tcgggtggtg
     3361 ctgatgtacc gcttcgagga ggagctgttc ctgcgcagcc tgcaagacta caaaattcag
     3421 tctgccctgc tggtgccaac cctgttcagc ttcttcgcta agagcaccct gatcgacaag
     3481 tacgacctgt ctaacctgca cgagattgcc tctggcggcg ccccactgtc taaggaggtg
     3541 ggcgaagccg tggccaagcg ctttcatctg ccaggcatcc gccagggcta cggcctgacc
     3601 gagacaacca gcgccattct gattacccca gagggcgacg acaagcctgg cgccgtgggc
     3661 aaggtggtgc cattcttcga ggccaaggtg gtggacctgg acaccggcaa gaccctggga
     3721 gtgaaccagc gcggcgagct gtgtgtgcgc ggccctatga ttatgtccgg ctacgtgaat
     3781 aaccctgagg ccacaaacgc cctgatcgac aaggacggct ggctgcactc tggcgacatt
     3841 gcctactggg acgaggacga gcacttcttc atcgtggacc gcctgaagtc tctgatcaag
     3901 tacaagggct accaggtggc cccagccgag ctggagtcta tcctgctgca gcaccctaac
     3961 attttcgacg ccggagtggc cggcctgccc gacgacgatg ccggcgagct gcctgccgcc
     4021 gtcgtcgtgc tggaacacgg caagaccatg accgagaagg agatcgtgga ctatgtggcc
     4081 agccaggtga caaccgccaa gaagctgcgc ggcggagtgg tgttcgtgga cgaggtgccc
     4141 aagggcctga ccggcaagct ggacgcccgc aagatccgcg agatcctgat caaggctaag
     4201 aaaggcggca agatcgccgt gtaataattc tagagtcggg gcggccggcc gcttcgagca
     4261 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4321 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4381 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     4441 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccaggt
     4501 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4561 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4621 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4681 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4741 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4801 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4861 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4921 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4981 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5041 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5101 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5161 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5221 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5281 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5341 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5401 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5461 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5521 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5581 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5641 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5701 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5761 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5821 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5881 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5941 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6001 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6061 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6301 cacatggctc gac
//
