pDream2.1/MCS

LOCUS       pDream2.1 MCS                7159 bp ds-DNA     circular SYN 23-FEB-2013
DEFINITION  Vector for expression of N-terminally FLAG-tagged proteins in 
            bacterial, insect, and mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pDream2.1 MCS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7159)
  AUTHORS   GenScript
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, January 11, 2020 from SnapGene Viewer 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7159
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_recomb     181..1008
                     /note="baculovirus recombination region (lef2/ORF603)"
                     /note="contains ORF603 and part of lef2"
     CDS             181..305
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /note="lef2"
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(343..948)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /note="ORF603"
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1086..1389
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1390..1593
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        1615..1633
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1634..1658
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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        1661..1770
                     /note="p10 promoter"
                     /note="baculovirus promoter for expression in insect cells"
     protein_bind    1772..1796
                     /gene="mutant version of attB"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1810..1812
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             1813..1836
                     /codon_start=1
                     /product="FLAG epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     misc_feature    1837..1882
                     /note="MCS"
                     /note="multiple cloning site"
     protein_bind    1905..1929
                     /gene="mutant version of attB"
                     /bound_moiety="BP clonase(TM)"
                     /note="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1931..2407
                     /note="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     terminator      2408..2455
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2548..2976
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2990..3319
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3170..3305
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3386..4180
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4354..4475
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     polyA_site      4519..4574
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     misc_recomb     4722..5427
                     /note="baculovirus recombination region (ORF1629)"
                     /note="contains part of ORF1629"
     CDS             complement(4839..5427)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /note="ORF1629"
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      complement(5637..6224)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6237..7097)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
       61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
      121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
      181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
      241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
      301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
      361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
      421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
      481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
      541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
      601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
      661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
      721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
      781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
      841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
      901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
      961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggccc
     1021 tcgagtagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     1081 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     1141 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     1201 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     1261 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     1321 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     1381 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     1441 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     1501 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     1561 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtgg atcttaatac
     1621 gactcactat aggggaattg tgagcggata acaattccgg gacctttaat tcaacccaac
     1681 acaatatatt atagttaaat aagaattatt atcaaatcat ttgtatatta attaaaatac
     1741 tatactgtaa attacatttt atttacaatc aacaagtttg tacaaaaaag caggctaagg
     1801 agatatacca tggattacaa ggatgacgac gataagggat ccgagatctg gaattcggta
     1861 ccgagctcaa gcttgcggcc gcctatagtg tcacctaaat cgtaacccag ctttcttgta
     1921 caaagtggtc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     1981 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2041 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2101 tgggatgaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     2161 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     2221 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     2281 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     2341 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     2401 tgtccttcta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtctag
     2461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2521 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2881 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2941 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3241 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3301 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3361 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3421 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3481 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3541 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3601 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3661 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3721 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3781 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3841 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3901 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3961 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4021 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4081 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4141 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4201 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4261 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4321 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgttcatg aagccccttg
     4501 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4561 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4621 agtatttggt ttagagtttg gcaacatatg cccatatgta actagcataa ccccttgggg
     4681 cctctaaacg ggtcttgagg ggttttttgc tgaaagcatg cggaggaaat tctccttgaa
     4741 gtttccctgg tgttcaaagt aaaggagttt gcaccagacg cacctctgtt cactggtccg
     4801 gcgtattaaa acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt
     4861 tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta
     4921 gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat
     4981 attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt
     5041 tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa
     5101 tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt
     5161 tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac
     5221 aattgactcg acgtaaacac gttaaataga gcttggacat atttaacatc gggcgtgtta
     5281 gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa
     5341 gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc
     5401 aaatttgtag ttgagctttt tggaattgcg atcgcataac ttcgtatagc atacattata
     5461 cgaagttata agctcggaac gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5641 cataggctcc gccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5821 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgttac
     6241 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6301 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6361 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6421 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6481 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6541 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6601 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6661 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6721 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6781 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6841 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6901 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6961 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7021 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7081 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7141 tgtctcatgt ccgcgcgtt
//