LOCUS       Exported                6721 bp ds-DNA     circular SYN 26-JUN-2025
DEFINITION  Gateway(R) destination vector for expressing C-terminally V5- and 
            6xHis-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  AUTHORS   Thermo Fisher (Invitrogen)
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 27, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6721
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     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"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..918
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    956..2152
                     /label=SIRT3
     protein_bind    complement(2186..2193)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2219..2260
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2270..2287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2316..2540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2586..3014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3028..3357
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3208..3343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3424..4218
                     /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"
     polyA_signal    4394..4515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4966..5554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5725..6585)
                     /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"
     promoter        complement(6586..6690)
                     /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 gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatggc
      961 gttctggggt tggcgcgccg cggcagccct ccggctgtgg ggccgggtag ttgaacgggt
     1021 cgaggccggg ggaggcgtgg ggccgtttca ggcctgcggc tgtcggctgg tgcttggcgg
     1081 cagggacgat gtgagtgcgg ggctgagagg cagccatggg gcccgcggtg agcccttgga
     1141 cccggcgcgc cccttgcaga ggcctcccag acccgaggtg cccagggcat tccggaggca
     1201 gccgagggca gcagctccca gtttcttctt ttcgagtatt aaaggtggaa gaaggtccat
     1261 atctttttct gtgggtgctt caagtgttgt tggaagtgga ggcagcagtg acaaggggaa
     1321 gctttccctg caggatgtag ctgagctgat tcgggccaga gcctgccaga gggtggtggt
     1381 catggtgggg gccggcatca gcacacccag tggcattcca gacttcagat cgccggggag
     1441 tggcctgtac agcaacctcc agcagtacga tctcccgtac cccgaggcca tttttgaact
     1501 cccattcttc tttcacaacc ccaagccctt tttcactttg gccaaggagc tgtaccctgg
     1561 aaactacaag cccaacgtca ctcactactt tctccggctg cttcatgaca aggggctgct
     1621 tctgcggctc tacacgcaga acatcgatgg gcttgagaga gtgtcgggca tccctgcctc
     1681 aaagctggtt gaagctcatg gaacctttgc ctctgccacc tgcacagtct gccaaagacc
     1741 cttcccaggg gaggacattc gggctgacgt gatggcagac agggttcccc gctgcccggt
     1801 ctgcaccggc gttgtgaagc ccgacattgt gttctttggg gagccgctgc cccagaggtt
     1861 cttgctgcat gtggttgatt tccccatggc agatctgctg ctcatccttg ggacctccct
     1921 ggaggtggag ccttttgcca gcttgaccga ggccgtgcgg agctcagttc cccgactgct
     1981 catcaaccgg gacttggtgg ggcccttggc ttggcatcct cgcagcaggg acgtggccca
     2041 gctgggggac gtggttcacg gcgtggaaag cctagtggag cttctgggct ggacagaaga
     2101 gatgcgggac cttgtgcagc gggaaactgg gaagcttgat ggaccagaca aaaagggtgg
     2161 gcgcgccgac ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg
     2221 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     2281 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2341 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2401 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2461 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2521 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2581 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2641 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2881 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2941 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3001 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3121 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3181 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3241 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3301 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3361 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3421 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3481 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3541 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3601 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3661 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3721 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3781 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3841 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3901 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3961 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4021 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4081 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4141 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4201 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4261 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4321 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4381 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4441 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4501 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4561 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4621 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4681 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4741 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4801 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4861 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4921 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4981 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5041 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5101 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5161 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5221 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5281 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5341 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5401 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5461 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5521 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5581 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5641 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5701 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5761 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5821 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5881 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5941 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6001 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6061 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6121 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6181 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6241 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6301 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6361 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6421 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6481 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6541 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6601 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6661 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6721 c
//
