LOCUS       40924_11660        5802 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Control vector with a strong constitutive CMV promoter for 
            expressing secreted Gaussia luciferase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5802)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5802)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5802
                     /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             920..1474
                     /codon_start=1
                     /label=hGLuc
                     /note="secreted Gaussia luciferase"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    1486..1534
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     rep_origin      1668..2096
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2110..2440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2507..3298
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3475..3608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3645..3661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3669..3685)
                     /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(3693..3723)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3738..3759)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4047..4635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4809..5666)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5667..5771)
                     /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 ccagccacca tgggagtcaa agttctgttt gccctgatct gcatcgctgt
      961 ggccgaggcc aagcccaccg agaacaacga agacttcaac atcgtggccg tggccagcaa
     1021 cttcgcgacc acggatctcg atgctgaccg cgggaagttg cccggcaaga agctgccgct
     1081 ggaggtgctc aaagagatgg aagccaatgc ccggaaagct ggctgcacca ggggctgtct
     1141 gatctgcctg tcccacatca agtgcacgcc caagatgaag aagttcatcc caggacgctg
     1201 ccacacctac gaaggcgaca aagagtccgc acagggcggc ataggcgagg cgatcgtcga
     1261 cattcctgag attcctgggt tcaaggactt ggagcccatg gagcagttca tcgcacaggt
     1321 cgatctgtgt gtggactgca caactggctg cctcaaaggg cttgccaacg tgcagtgttc
     1381 tgacctgctc aagaagtggc tgccgcaacg ctgtgcgacc tttgccagca agatccaggg
     1441 ccaggtggac aagatcaagg gggccggtgg tgactaagcg gccgcaataa aatatcttta
     1501 ttttcattac atctgtgtgt tggttttttg tgtgtctaga aataattctt actgtcatgc
     1561 caagtaagat gcttttctgt gctgcaatag caggcatgct ggggatgcgg tgggctctat
     1621 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1681 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1741 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1801 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     1861 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1921 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1981 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     2041 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2101 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     2161 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2281 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2341 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2401 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2461 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2521 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2581 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2641 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2701 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2761 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2821 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2881 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2941 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3001 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3061 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3121 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3181 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3241 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3301 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3361 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3421 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3601 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     4261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
