LOCUS       Exported                7092 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 7092)
  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..7092
                     /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..1888
                     /label=SIRT5
     protein_bind    complement(1922..1929)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1940..2659
                     /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    2687..2911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2957..3385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3399..3728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3579..3714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3795..4589
                     /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    4765..4886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5337..5925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6096..6956)
                     /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(6957..7061)
                     /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 gcgacctctc cagattgtcc caagtcgatt gatttcccag ctatattgtg gcctgaagcc
     1021 tccagcgtcc acacgaaacc agatttgcct gaaaatggct cggccaagtt caagtatggc
     1081 agattttcga aagttttttg caaaagcaaa gcacatagtc atcatctcag gagctggtgt
     1141 tagtgcagaa agtggtgttc cgaccttcag aggagctgga ggttattgga gaaaatggca
     1201 agcccaggac ctggcgactc ccctggcctt tgcccacaac ccgtcccggg tgtgggagtt
     1261 ctaccactac cggcgggagg tcatggggag caaggagccc aacgccgggc accgcgccat
     1321 agccgagtgt gagacccggc tgggcaagca gggccggcga gtcgtggtca tcacccagaa
     1381 catcgatgag ctgcaccgca aggctggcac caagaacctt ctggagatcc atggtagctt
     1441 atttaaaact cgatgtacct cttgtggagt tgtggctgag aattacaaga gtccaatttg
     1501 tccagcttta tcaggaaaag gtgctccaga acctggaact caagatgcca gcatcccagt
     1561 tgagaaactt ccccggtgtg aagaggcagg ctgcgggggc ttgctgcgac ctcacgtcgt
     1621 gtggtttgga gaaaacctgg atcctgccat tctggaggag gttgacagag agctcgccca
     1681 ctgtgattta tgtctagtgg tgggcacttc ctctgtggtg tacccagcag ccatgtttgc
     1741 cccccaggtg gctgccaggg gcgtgccagt ggctgaattt aacacggaga ccaccccagc
     1801 tacgaacaga ttcaggtttc atttccaggg accctgtgga acgactcttc ctgaagccct
     1861 tgcctgtcat gaaaatgaaa ctgtttctaa gggtgggcgc gccgacccag ctttcttgta
     1921 caaagtggtt cgatctagaa tggctagcaa aggagaagaa cttttcactg gagttgtccc
     1981 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
     2041 tgaaggtgat gctacatacg gaaagcttac ccttaaattt atttgcacta ctggaaaact
     2101 acctgttcca tggccaacac ttgtcactac tttctcttat ggtgttcaat gcttttcccg
     2161 ttatccggat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
     2221 acaggaacgc actatatctt tcaaagatga cgggaactac aagacgcgtg ctgaagtcaa
     2281 gtttgaaggt gatacccttg ttaatcgtat cgagttaaaa ggtattgatt ttaaagaaga
     2341 tggaaacatt ctcggacaca aactcgagta caactataac tcacacaatg tatacatcac
     2401 ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca acattgaaga
     2461 tggatccgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     2521 ccttttacca gacaaccatt acctgtcgac acaatctgcc ctttcgaaag atcccaacga
     2581 aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta cacatggcat
     2641 ggatgagctc tacaaataat gaattaaacc cgctgatcag cctcgactgt gccttctagt
     2701 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2761 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2821 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2881 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2941 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3301 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3361 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3481 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3661 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3721 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3781 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3841 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3901 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3961 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4021 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4081 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4141 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4201 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4261 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4321 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4381 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4441 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4501 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4561 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     4621 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4681 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4741 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4801 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4861 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4921 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4981 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5041 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5101 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5161 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5221 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5281 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5341 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5401 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5461 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5521 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5581 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5641 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5701 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5761 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5821 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5881 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5941 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6001 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6061 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6121 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6181 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6241 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6301 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6361 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6421 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6481 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6541 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6601 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6661 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6721 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6781 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6841 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6901 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6961 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7021 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7081 ccacctgacg tc
//
