LOCUS       40924_43478        6730 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector for constitutive low-level expression of secreted 
            Cypridina luciferase under control of the HSV TK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  AUTHORS   Thermo Scientific (Pierce)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6730)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..777
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             790..2448
                     /codon_start=1
                     /label=CLuc
                     /note="secreted Cypridina luciferase"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     polyA_signal    2474..2585
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2708..3037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        3085..3132
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3151..3747
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3880..4013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4057..4914)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5178..5766
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6052
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6508..6524
                     /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    6639..6730
                     /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 tgaagaccct gatcctggcc gtggccctgg tgtactgcgc caccgtgcac
      841 tgccaggact gcccctacga gcccgacccc cccaataccg tgcctaccag ctgcgaggcc
      901 aaagagggcg aatgcatcga cagcagctgc ggcacctgta cccgggacat cctgagcgac
      961 ggcctgtgcg agaacaagcc cggcaagacc tgctgccgga tgtgccagta cgtgatcgag
     1021 tgccgggtgg aagccgccgg atggttccgg accttctacg gcaagcggtt ccagtttcag
     1081 gaacccggca cctacgtcct gggccagggc acaaagggcg gcgactggaa ggtgtccatc
     1141 accctggaaa acctggacgg caccaagggc gccgtgctga ccaagaccag actggaagtg
     1201 gccggcgaca tcatcgatat cgcccaggcc accgagaacc ccatcaccgt gaacggcgga
     1261 gccgacccca tcattgccaa cccctacacc atcggcgaag tgacaatcgc tgtggtggaa
     1321 atgcccggct tcaatatcac cgtcatcgag ttcttcaagc tgatcgtgat cgacatcctg
     1381 ggcggcagaa gcgtgcggat cgcccccgat accgccaaca agggcatgat cagcggcctg
     1441 tgtggcgacc tgaagatgat ggaagatacc gacttcacca gcgaccccga gcagctggcc
     1501 atccagccca agatcaacca ggaatttgac ggctgccccc tgtacggcaa ccccgacgac
     1561 gtggcctact gcaagggcct gctggaaccc tacaaggaca gctgcagaaa ccccatcaac
     1621 ttctactact acaccatcag ctgcgccttc gcccggtgca tgggcggaga tgagcgcgct
     1681 agccacgtgc tgctggacta cagagagaca tgcgccgctc ccgagacacg gggcacatgt
     1741 gtgctgagcg gccacacctt ctacgacacc ttcgacaagg ccagatacca gttccagggc
     1801 ccctgcaaag aaatcctgat ggccgccgac tgcttctgga acacctggga cgtgaaggtg
     1861 tcccaccgga acgtggacag ctataccgag gtggaaaaag tgcggatcag aaagcagagc
     1921 accgtggtcg agctgattgt ggacggcaag cagatcctcg tgggcggcga ggccgtgtcc
     1981 gtgccctaca gcagccagaa caccagcatc tactggcagg acggcgacat cctgaccacc
     2041 gccatcctgc ctgaggccct ggtggtcaag ttcaacttca agcagctgct ggtggtgcac
     2101 atccgggacc ccttcgacgg caagacatgc ggcatctgcg gcaactacaa ccaggacttc
     2161 agcgacgaca gcttcgacgc cgagggcgcc tgcgacctga cccctaatcc tcccggctgc
     2221 accgaggaac agaagcccga ggccgagcgg ctgtgcaata gcctgttcgc cggccagagc
     2281 gacctggacc agaaatgcaa cgtgtgccac aagcccgacc gggtggaacg gtgtatgtac
     2341 gagtactgcc tgcggggcca gcagggcttc tgcgatcacg cctgggagtt caagaaagag
     2401 tgctacatca agcacggcga caccctggaa gtgcccgacg agtgcaagtg agcggccgca
     2461 aaatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2521 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2581 gcatcacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2641 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     2701 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2761 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2881 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2941 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3001 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3061 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     3121 gtataatacg acaaggtgag gaactaaacc atgaccgagt acaagcccac ggtgcgcctc
     3181 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     3241 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     3301 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3361 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     3421 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     3481 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     3541 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     3601 gccgagcgcg ccggggtgcc cgccttcctg gagacatccg cgccccgcaa cctccccttc
     3661 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     3721 tggtgcatga cccgcaagcc cggtgcctga cacgtgctac gagatttcga ttccaccgcc
     3781 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3841 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3901 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3961 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4021 acctctagct agagcttggc gtaatcatgg tcattaccaa tgcttaatca gtgaggcacc
     4081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4141 aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac cgcgagaacc
     4201 acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta caggcatcgt
     4381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatattctt
     4921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4981 tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt acaaccaatt aaccaattct
     5041 gaacattatc gcgagcccat ttatacctga atatggctca taacacccct tgctcatgac
     5101 caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa
     5161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5221 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5341 tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5821 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5881 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5941 gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc cctgacggat
     6001 ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca gtcttaagct
     6061 cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc
     6121 cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc agaatattta
     6181 agggcgcctg tcactttgct tgatatatga gaattattta accttataaa tgagaaaaaa
     6241 gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta taattaaact
     6301 attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt aaagagaatt
     6361 aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca aaattataca
     6421 tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat ttattatcat
     6481 ttagaataaa ttttgtgtcg cccttaattg tgagcggata acaattacga gcttcatgca
     6541 cagtggcgtt gacattgatt attgactagc atgttctttc ctgcgttatc ccctgattct
     6601 gtggataacc gtattaccgc catgcattag ttattaataa catacgctct ccatcaaaac
     6661 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     6721 acatttctct
//
