LOCUS       sequence-8829-ex        7755 bp    DNA     circular SYN 15-DEC-2025
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    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            J. 2008 Mar . 3(3):370-7."
COMMENT     Sequence Label: pEZYmyc-His
FEATURES             Location/Qualifiers
     source          1..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    950..1074
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1503..1808)
                     /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(1605..1624)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2153..2812)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2727..2746
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2813..2915)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3093..3217)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3240..3269
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3285..3302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3346..3363)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3352..3576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3622..4050
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3709..3728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3919..3940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4059..4079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4064..4393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4244..4379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4306..4325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4460..5254
                     /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(4514..4533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5124..5143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5428..5549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5465..5484)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5519..5538
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5598..5614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5598..5614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5611..5633)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5622..5638
                     /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(join(5646..5652,5653..5670,5671..5676))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5691..5712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5829..5846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6000..6588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6080..6099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6759..7550,7551..7619))
                     /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     7382..7401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7620..7724)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccatgcaa ttgattaata caagtttgta
      961 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     1021 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatgaatcaa
     1081 ctacttagat ggtattagtg acctgtagtc gaccgacagc cttccaaatg ttcttcgggt
     1141 gatgctgcca acttagtcga ccgacagcct tccaaatgtt cttctcaaac ggaatcgtcg
     1201 tatccagcct actcgctatt gtcctcaatg ccgtattaaa tcataaaaag aaataagaaa
     1261 aagaggtgcg agcctctttt ttgtgtgaca aaataaaaac atctacctat tcatatacgc
     1321 tagtgtcata gtcctgaaaa tcatctgcat caagaacaat ttcacaactc ttatactttt
     1381 ctcttacaag tcgttcggct tcatctggat tttcagcctc tatacttact aaacgtgata
     1441 aagtttctgt aatttctact gtatcgacct gcagactggc tgtgtataag ggagcctgac
     1501 atttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg
     1561 agatcagcca cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc
     1621 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct
     1681 gacagcagac gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata
     1741 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta
     1801 aactgcattt caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg
     1861 ggcgacctca gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg
     1921 ggcttcattc tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg
     1981 agcaactgat agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc
     2041 tttttgacat acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc
     2101 gcaaatacgc atactgttat ctggctttta gtaagccgga tccacgcggc gtttacgccc
     2161 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     2221 catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     2281 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     2341 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     2401 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     2461 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     2521 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     2581 tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     2641 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     2701 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
     2761 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     2821 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     2881 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
     2941 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
     3001 ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttagtcgact
     3061 acaggtcact aataccatct aagtagttga ttcatagtga ctggatatgt tgtgttttac
     3121 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     3181 tttacgtttc tcgttcagct ttcttgtaca aagtggtcct cgaggtacca agctttctag
     3241 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc
     3301 attgagttta aacggtctcc agcttaagtt taaaccgctg atcagcctcg actgtgcctt
     3361 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3421 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3481 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3541 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3601 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3661 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3721 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3781 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3841 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3901 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3961 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4021 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4081 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4141 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4321 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4381 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4441 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4501 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4561 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4621 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4681 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4741 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4801 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4861 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4921 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4981 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5041 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5101 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5161 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5221 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5281 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5341 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5401 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5461 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5521 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5581 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5641 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5701 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5761 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5821 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5881 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5941 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6001 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6061 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6121 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6181 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6241 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6301 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6361 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6421 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6481 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6541 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6601 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6661 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6721 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6781 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6841 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6901 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6961 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7021 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7081 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7141 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7201 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7261 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7321 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7381 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7441 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7501 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7561 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7621 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7681 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7741 gtgccacctg acgtc
//
