LOCUS       Exported                7754 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pcDNA3.1(-)/Myc-His B converted Gateway Destination vector for 
            expressing C-terminal myc-His tagged fusion proteins in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    pEZYmyc-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7754)
  AUTHORS   Guo F, Chiang MY, Wang Y, Zhang YZ
  TITLE     An in vitro recombination method to convert restriction- and 
            ligation-independent expression vectors.
  JOURNAL   Biotechnol J. 2008 Mar . 3(3):370-7.
  PUBMED    18064608
REFERENCE   2  (bases 1 to 7754)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7754
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    952..1076
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1505..1810)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1607..1626)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2160..2813)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     primer_bind     2728..2747
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2814..2916)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3094..3218)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3241..3270
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3286..3303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3347..3364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3353..3577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3623..4051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3710..3729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3920..3941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4060..4080)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4065..4394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4245..4380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4307..4326
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4461..5255
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4515..4534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5125..5144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5429..5550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5466..5485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5520..5539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5599..5615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5599..5615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5612..5634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5623..5639
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5647..5677)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5692..5713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5830..5847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6001..6589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6081..6100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6760..7620)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7383..7402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7621..7725)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac gggccctcta gactcgagcg gccgccatgc aattgattaa tacaagtttg
      961 tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
     1021 ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc actatgaatc
     1081 aactacttag atggtattag tgacctgtag tcgaccgaca gccttccaaa tgttcttcgg
     1141 gtgatgctgc caacttagtc gaccgacagc cttccaaatg ttcttctcaa acggaatcgt
     1201 cgtatccagc ctactcgcta ttgtcctcaa tgccgtatta aatcataaaa agaaataaga
     1261 aaaagaggtg cgagcctctt ttttgtgtga caaaataaaa acatctacct attcatatac
     1321 gctagtgtca tagtcctgaa aatcatctgc atcaagaaca atttcacaac tcttatactt
     1381 ttctcttaca agtcgttcgg cttcatctgg attttcagcc tctatactta ctaaacgtga
     1441 taaagtttct gtaatttcta ctgtatcgac ctgcagactg gctgtgtata agggagcctg
     1501 acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc
     1561 tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata tcggtggtca
     1621 tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg gagactttat
     1681 ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc gtgtcaataa
     1741 tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag gtgtaaacct
     1801 taaactgcat ttcaccagcc cctgttctcg tcagcaaaag agccgttcat ttcaataaac
     1861 cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc acgcagacga
     1921 cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc atatatgcct
     1981 tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc atagcatacc
     2041 tctttttgac atacttcggg tatacatatc agtatatatt cttataccgc aaaaatcagc
     2101 gcgcaaatac gcatactgtt atctggcttt tagtaagccg gatccacgcg gcgtttacgc
     2161 cccccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     2221 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     2281 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     2341 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     2401 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     2461 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     2521 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     2581 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     2641 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     2701 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     2761 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     2821 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     2881 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg
     2941 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     3001 tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttagtcgac
     3061 tacaggtcac taataccatc taagtagttg attcatagtg actggatatg ttgtgtttta
     3121 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     3181 ttttacgttt ctcgttcagc tttcttgtac aaagtggtcc tcgaggtacc aagctttcta
     3241 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
     3301 cattgagttt aaacggtctc cagcttaagt ttaaaccgct gatcagcctc gactgtgcct
     3361 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3421 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3481 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3541 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3601 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3661 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3841 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3901 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3961 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4021 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4141 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4201 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4261 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4321 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4381 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4441 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4501 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4561 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4621 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4681 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4741 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4801 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4861 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4921 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4981 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5041 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5101 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5161 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5221 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5281 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5341 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5401 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5461 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5521 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5581 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7741 agtgccacct gacg
//
