LOCUS       Exported                7329 bp ds-DNA     circular SYN 26-JUN-2025
DEFINITION  Gateway(R) destination vector for expressing C-terminally GFP-tagged
            proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7329)
  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..7329
                     /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    914..921
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    959..2125
                     /label=SIRT2
     protein_bind    complement(2159..2166)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2177..2896
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     polyA_signal    2924..3148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3194..3622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3636..3965
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3816..3951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4032..4826
                     /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    5002..5123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5574..6162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6333..7193)
                     /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(7194..7298)
                     /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 taagcttgat caaacaagtt tgtacaaaaa agcaggctcc gcggccgccc ccttcaccat
      961 ggcagagcca gacccctctc accctctgga gacccaggca gggaaggtgc aggaggctca
     1021 ggactcagat tcagactctg agggaggagc cgctggtgga gaagcagaca tggacttcct
     1081 gcggaactta ttctcccaga cgctcagcct gggcagccag aaggagcgtc tgctggacga
     1141 gctgaccttg gaaggggtgg cccggtacat gcagagcgaa cgctgtcgca gagtcatctg
     1201 tttggtggga gctggaatct ccacatccgc aggcatcccc gactttcgct ctccatccac
     1261 cggcctctat gacaacctag agaagtacca tcttccctac ccagaggcca tctttgagat
     1321 cagctatttc aagaaacatc cggaaccctt cttcgccctc gccaaggaac tctatcctgg
     1381 gcagttcaag ccaaccatct gtcactactt catgcgcctg ctgaaggaca aggggctact
     1441 cctgcgctgc tacacgcaga acatagatac cctggagcga atagccgggc tggaacagga
     1501 ggacttggtg gaggcgcacg gcaccttcta cacatcacac tgcgtcagcg ccagctgccg
     1561 gcacgaatac ccgctaagct ggatgaaaga gaagatcttc tctgaggtga cgcccaagtg
     1621 tgaagactgt cagagcctgg tgaagcctga tatcgtcttt tttggtgaga gcctcccagc
     1681 gcgtttcttc tcctgtatgc agtcagactt cctgaaggtg gacctcctcc tggtcatggg
     1741 tacctccttg caggtgcagc cctttgcctc cctcatcagc aaggcacccc tctccacccc
     1801 tcgcctgctc atcaacaagg agaaagctgg ccagtcggac cctttcctgg ggatgattat
     1861 gggcctcgga ggaggcatgg actttgactc caagaaggcc tacagggacg tggcctggct
     1921 gggtgaatgc gaccagggct gcctggccct tgctgagctc cttggatgga agaaggagct
     1981 ggaggacctt gtccggaggg agcacgccag catagatgcc cagtcggggg cgggggtccc
     2041 caaccccagc acttcagctt cccccaagaa gtccccgcca cctgccaagg acgaggccag
     2101 gacaacagag agggagaaac cccagaaggg tgggcgcgcc gacccagctt tcttgtacaa
     2161 agtggttcga tctagaatgg ctagcaaagg agaagaactt ttcactggag ttgtcccaat
     2221 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     2281 aggtgatgct acatacggaa agcttaccct taaatttatt tgcactactg gaaaactacc
     2341 tgttccatgg ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcccgtta
     2401 tccggatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     2461 ggaacgcact atatctttca aagatgacgg gaactacaag acgcgtgctg aagtcaagtt
     2521 tgaaggtgat acccttgtta atcgtatcga gttaaaaggt attgatttta aagaagatgg
     2581 aaacattctc ggacacaaac tcgagtacaa ctataactca cacaatgtat acatcacggc
     2641 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg
     2701 atccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     2761 tttaccagac aaccattacc tgtcgacaca atctgccctt tcgaaagatc ccaacgaaaa
     2821 gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga
     2881 tgagctctac aaataatgaa ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2941 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3001 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3061 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3121 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3181 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3241 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3301 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3361 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3421 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3481 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3541 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3601 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3661 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3781 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3841 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3901 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3961 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4021 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4081 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4141 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4201 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4261 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4321 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4381 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4441 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4501 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4561 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4621 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4681 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4741 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4801 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4861 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4921 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4981 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5041 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5101 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5161 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5221 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5281 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5341 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5401 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5461 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5521 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5581 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5641 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5701 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5761 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5821 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5881 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5941 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6001 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6061 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6121 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6181 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6241 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6301 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6361 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6421 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6481 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6541 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6601 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6661 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6721 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6781 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6841 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6901 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6961 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7021 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7081 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7141 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7201 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7261 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7321 cctgacgtc
//
