LOCUS       40924_22438        5028 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian vector with a minimal promoter for measuring promoter or 
            enhancer activity using secreted Gaussia luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5028)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5028)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..131
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             146..700
                     /codon_start=1
                     /label=hGLuc
                     /note="secreted Gaussia luciferase"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    712..760
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     rep_origin      894..1322
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1336..1666
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1733..2524
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    2701..2834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2871..2887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2895..2911)
                     /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(2919..2949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2964..2985)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3273..3861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4035..4892)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4893..4997)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcttgg aattctgcag atatcctcga gcccaagctt ggtaccgagc
       61 tcggatcctt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc
      121 gacccgctta aaagatccag ccaccatggg agtcaaagtt ctgtttgccc tgatctgcat
      181 cgctgtggcc gaggccaagc ccaccgagaa caacgaagac ttcaacatcg tggccgtggc
      241 cagcaacttc gcgaccacgg atctcgatgc tgaccgcggg aagttgcccg gcaagaagct
      301 gccgctggag gtgctcaaag agatggaagc caatgcccgg aaagctggct gcaccagggg
      361 ctgtctgatc tgcctgtccc acatcaagtg cacgcccaag atgaagaagt tcatcccagg
      421 acgctgccac acctacgaag gcgacaaaga gtccgcacag ggcggcatag gcgaggcgat
      481 cgtcgacatt cctgagattc ctgggttcaa ggacttggag cccatggagc agttcatcgc
      541 acaggtcgat ctgtgtgtgg actgcacaac tggctgcctc aaagggcttg ccaacgtgca
      601 gtgttctgac ctgctcaaga agtggctgcc gcaacgctgt gcgacctttg ccagcaagat
      661 ccagggccag gtggacaaga tcaagggggc cggtggtgac taagcggccg caataaaata
      721 tctttatttt cattacatct gtgtgttggt tttttgtgtg tctagaaata attcttactg
      781 tcatgccaag taagatgctt ttctgtgctg caatagcagg catgctgggg atgcggtggg
      841 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
      901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1021 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     1081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1261 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1321 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg
     1381 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     1441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     1501 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1561 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     1621 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     1681 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1741 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1801 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1861 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1921 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1981 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2041 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2101 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2161 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2221 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     2281 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2341 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2401 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2461 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2521 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2581 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2641 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2821 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     2881 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2941 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3001 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3061 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3241 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     3481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4141 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4981 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
