LOCUS       pGLuc_Mini-TK_2         5028 bp    DNA     circular     24-SEP-2012
DEFINITION  Cloning vector pGLuc_Mini-TK_2, complete sequence.
ACCESSION
VERSION
KEYWORDS    .
SOURCE      Cloning vector pGLuc_Mini-TK_2
  ORGANISM  Cloning vector pGLuc_Mini-TK_2
            Unclassified.
REFERENCE   1  (bases 1 to 5028)
  AUTHORS   Tzertzinis,G.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-SEP-2012) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA
FEATURES             Location/Qualifiers
     source          1..5028
                     /organism="Cloning vector pGLuc_Mini-TK_2"
                     /mol_type="other DNA"
     misc_feature    12..68
                     /note="MCS (BglII-BamHI)"
     promoter        69..131
                     /note="minimal thymidine kinase promoter (PminiTK)"
     misc_RNA        140..149
                     /note="Kozak consensus sequence for GLuc"
     gene            146..703
                     /gene="GLuc"
     CDS             146..703
                     /gene="GLuc"
                     /codon_start=1
                     /product="Gaussia princeps luciferase"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADR
                     GKLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKE
                     SAQGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKW
                     LPQRCATFASKIQGQVDKIKGAGGD"
     sig_peptide     146..196
                     /gene="GLuc"
                     /note="secretory signal sequence"
     polyA_signal    712..760
                     /note="synthetic polyA"
     promoter        1346..1681
                     /note="SV40 promoter (clockwise)"
     gene            1733..2527
                     /gene="aph(3')-II"
     CDS             1733..2527
                     /gene="aph(3')-II"
                     /note="neoR, kanR, nptII (confers resistance to neomycin
                     and kanamycin)"
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(3273..3861)
                     /note="pUC19 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
     gene            complement(4032..4892)
                     /gene="bla"
     CDS             complement(4032..4892)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
BASE COUNT     1153 a   1328 c   1311 g   1236 t
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
//
