LOCUS       40924_11611        6905 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector with a strong constitutive CMV promoter for 
            expressing secreted Cypridina luciferase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6905)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             919..2577
                     /codon_start=1
                     /label=CLuc
                     /note="secreted Cypridina luciferase"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     polyA_signal    2589..2637
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     rep_origin      2771..3199
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3213..3543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3610..4401
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4578..4711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4748..4764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4772..4788)
                     /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)."
     promoter        complement(4796..4826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4841..4862)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5150..5738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5912..6769)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6770..6874)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccgccaccat gaagacctta attcttgccg ttgcattagt ctactgcgcc
      961 actgttcatt gccaggactg tccttacgaa cctgatccac caaacacagt tccaacttcc
     1021 tgtgaagcta aagaaggaga atgtattgat agcagctgtg gcacctgcac gagagacata
     1081 ctatcagatg gactgtgtga aaataaacca ggaaaaacat gttgccgaat gtgtcagtat
     1141 gtaattgaat gcagagtaga ggccgcagga tggtttagaa cattctatgg aaagagattc
     1201 cagttccagg aacctggtac atacgtgttg ggtcaaggaa ccaagggcgg cgactggaag
     1261 gtgtccatca ccctggagaa cctggatgga accaaggggg ctgtgctgac caagacaaga
     1321 ctggaagtgg ctggagacat cattgacatc gctcaagcta ctgagaatcc catcactgta
     1381 aacggtggag ctgaccctat catcgccaac ccgtacacca tcggcgaggt caccatcgct
     1441 gttgttgaga tgccaggctt caacatcacc gtcattgagt tcttcaaact gatcgtgatc
     1501 gacatcctcg gaggaagatc tgtaagaatc gccccagaca cagcaaacaa aggaatgatc
     1561 tctggcctct gtggagatct taaaatgatg gaagatacag acttcacttc agatccagaa
     1621 caactcgcta ttcagcctaa gatcaaccag gagtttgacg gttgtccact ctatggaaat
     1681 cctgatgacg ttgcatactg caaaggtctt ctggagccgt acaaggacag ctgccgcaac
     1741 cccatcaact tctactacta caccatctcc tgcgccttcg cccgctgtat gggtggagac
     1801 gagcgagcct cacacgtgct gcttgactac agggagacgt gcgctgctcc cgaaactaga
     1861 ggaacctgcg ttttgtctgg acatactttc tacgatacat ttgacaaagc aagataccaa
     1921 ttccagggtc cctgcaagga gattcttatg gccgccgact gtttctggaa cacttgggat
     1981 gtgaaggttt cacacaggaa tgttgactct tacactgaag tagagaaagt acgaatcagg
     2041 aaacaatcga ctgtagtaga actcattgtt gatggaaaac agattctggt tggaggagaa
     2101 gccgtgtccg tcccgtacag ctctcagaac acttccatct actggcaaga tggtgacata
     2161 ctgactacag ccatcctacc tgaagctctg gtggtcaagt tcaacttcaa gcaactgctc
     2221 gtcgtacata ttagagatcc attcgatggt aagacttgcg gtatttgcgg taactacaac
     2281 caggatttca gtgatgattc ttttgatgct gaaggagcct gtgatctgac ccccaaccca
     2341 ccgggatgca ccgaagaaca gaaacctgaa gctgaacgac tctgcaatag tctcttcgcc
     2401 ggtcaaagtg atcttgatca gaaatgtaac gtgtgccaca agcctgaccg tgtcgaacga
     2461 tgcatgtacg agtattgcct gaggggacaa cagggtttct gtgaccacgc atgggagttc
     2521 aagaaagaat gctacataaa gcatggagac accctagaag taccagatga atgcaaatag
     2581 gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgtct
     2641 agaaataatt cttactgtca tgccaagtaa gatgcttttc tgtgctgcaa tagcaggcat
     2701 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2761 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2821 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2881 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     2941 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3001 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3061 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3121 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3181 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3241 caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3481 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3541 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3601 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3661 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3721 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3781 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3841 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3901 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3961 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4021 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4081 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4141 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4201 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4261 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4321 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4381 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4441 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4501 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4561 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4621 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4681 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5341 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5581 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5701 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5761 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5821 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6901 acgtc
//
