LOCUS       40924_43498        6004 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive low-level expression of 
            intracellular green Renilla luciferase under control of the HSV TK 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6004)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6004)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..777
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             790..1722
                     /codon_start=1
                     /label=Green Renilla Luciferase
                     /note="green-emitting variant of Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGSALAFHYAYEHQDRIKAIVHMESVVDVIESWMGWPDIEEEL
                     ALIKSEEGEKMVLENNFFVETLLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1748..1859
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1982..2311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2359..2406
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2425..3021
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3154..3287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3331..4188)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4452..5040
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5310..5326
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5782..5798
                     /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)."
     misc_feature    5913..6004
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 actagtctcg aggaattcca agcttaatga gtcttcggac ctcgcggggg ccgcttaagc
       61 ggtggttagg gtttgtctga cgcgggggga gggggaagga acgaaacact ctcattcgga
      121 ggcggctcgg ggtttggtct tggtggccac gggcacgcag aagagcgccg cgatcctctt
      181 aagcaccccc ccgccctccg tggaggcggg ggtttggtcg gcgggtggta actggcgggc
      241 cgctgactcg ggcgggtcgc gcgccccaga gtgtgacctt ttcggtctgc tcgcagaccc
      301 ccgggcggcg ccgccgcggc ggcgacgggc tcgctgggtc ctaggctcca tggggaccgt
      361 atacgtggac aggctctgga gcatccgcac gactgcggtg atattaccgg agaccttctg
      421 cgggacgagc cgggtcacgc ggctgacgcg gagcgtccgt tgggcgacaa acaccaggac
      481 ggggcacagg tacactatct tgtcacccgg aggcgcgagg gactgcagga gcttcaggga
      541 gtggcgcagc tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga
      601 agaaatatat ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc
      661 attggcgaat tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca
      721 tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaagga
      781 tccgccacca tggccagcaa ggtgtacgac cccgagcagc ggaagcggat gatcaccggc
      841 cctcagtggt gggctcggtg caagcagatg aacgtgctgg acagcttcat caactactac
      901 gacagcgaga agcacgccga gaacgccgtg atcttcctgc acggcaacgc caccagcagc
      961 tacctgtggc ggcacgtggt gccccacatc gagcctgtgg ccagatgcat catccccgac
     1021 ctgatcggca tgggcaagag cggcaagtcc ggcaacggca gctaccggct gctggaccac
     1081 tacaagtacc tgaccgcttg gtttgagctg ctgaacctgc ccaagaagat catcttcgtc
     1141 ggccacgact ggggcagcgc cctggccttt cactacgcct acgagcacca ggaccggatc
     1201 aaggccatcg tgcacatgga aagcgtggtg gacgtgatcg agagctggat gggctggccc
     1261 gacatcgagg aagaactggc cctgatcaag agcgaagagg gcgagaagat ggtgctggaa
     1321 aacaacttct tcgtggaaac cctgctgccc agcaagatca tgcggaagct ggaacccgaa
     1381 gagttcgccg cctacctgga acccttcaaa gaaaagggcg aagtgcggag gcccaccctg
     1441 agctggccca gagagatccc cctggtcaag ggcggcaagc ccgacgtggt gcagatcgtg
     1501 cggaactaca acgcctacct gcgggccagc gacgacctgc ctaagctgtt catcgagagc
     1561 gaccccggct tcttcagcaa cgccatcgtg gaaggcgcca agaagttccc caacaccgag
     1621 ttcgtgaaag tgaagggcct gcacttcctc caggaagatg cccccgacga gatgggcaag
     1681 tacatcaaga gcttcgtgga acgggtgctg aagaacgagc agtgagcggc cgcaaaatca
     1741 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1801 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatca
     1861 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1921 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     1981 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2041 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2101 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2161 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2221 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2281 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2341 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     2401 tacgacaagg tgaggaacta aaccatgacc gagtacaagc ccacggtgcg cctcgccacc
     2461 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     2521 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     2581 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     2641 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     2701 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     2761 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     2821 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     2881 cgcgccgggg tgcccgcctt cctggagaca tccgcgcccc gcaacctccc cttctacgag
     2941 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     3001 atgacccgca agcccggtgc ctgacacgtg ctacgagatt tcgattccac cgccgccttc
     3061 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     3121 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     3181 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3241 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     3301 agctagagct tggcgtaatc atggtcatta ccaatgctta atcagtgagg cacctatctc
     3361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3421 gatacgggag ggcttaccat ctggccccag cgctgcgatg ataccgcgag aaccacgctc
     3481 accggctccg gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc
     3661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatat tcttcctttt
     4201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4261 tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat
     4321 tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgctca tgaccaaaat
     4381 cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     4441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4621 gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5101 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5161 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5221 ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac ggatggcctt
     5281 tttgcgtttc tacaaactct ttctgtgttg taaaacgacg gccagtctta agctcgggcc
     5341 ccctgggcgg ttctgataac gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc
     5401 ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcagaata tttaagggcg
     5461 cctgtcactt tgcttgatat atgagaatta tttaacctta taaatgagaa aaaagcaacg
     5521 cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta aactattcat
     5581 ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag aattaagaaa
     5641 ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta tacatgtcaa
     5701 cgataataca aaatataata caaactataa gatgttatca gtatttatta tcatttagaa
     5761 taaattttgt gtcgccctta attgtgagcg gataacaatt acgagcttca tgcacagtgg
     5821 cgttgacatt gattattgac tagcatgttc tttcctgcgt tatcccctga ttctgtggat
     5881 aaccgtatta ccgccatgca ttagttatta ataacatacg ctctccatca aaacaaaacg
     5941 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
     6001 ctct
//
