LOCUS       Exported                8403 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 8403)
  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..8403
                     /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"
     CDS             959..3199
                     /codon_start=1
                     /product="human NAD+-dependent protein deacetylase 
                     sirtuin-1"
                     /label=SIRT1
                     /note="homolog of budding yeast Sir2"
                     /translation="MADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLE
                     RSPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNG
                     PGLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESD
                     EEDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQ
                     IVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYA
                     RLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLL
                     RNYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCP
                     ADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHE
                     VPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRT
                     QKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCME
                     EKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISR
                     RLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESE
                     IEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
     protein_bind    complement(3233..3240)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3251..3970
                     /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    3998..4222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4268..4696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4710..5039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4890..5025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5106..5900
                     /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    6076..6197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6648..7236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7407..8267)
                     /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(8268..8372)
                     /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 ggcggacgag gcggccctcg cccttcagcc cggcggctcc ccctcggcgg cgggggccga
     1021 cagggaggcc gcgtcgtccc ccgccgggga gccgctccgc aagaggccgc ggagagatgg
     1081 tcccggcctc gagcggagcc cgggcgagcc cggtggggcg gccccagagc gtgaggtgcc
     1141 ggcggcggcc aggggctgcc cgggtgcggc ggcggcggcg ctgtggcggg aggcggaggc
     1201 agaggcggcg gcggcaggcg gggagcaaga ggcccaggcg actgcggcgg ctggggaagg
     1261 agacaatggg ccgggcctgc agggcccatc tcgggagcca ccgctggccg acaacttgta
     1321 cgacgaagac gacgacgacg agggcgagga ggaggaagag gcggcggcgg cggcgattgg
     1381 gtaccgagat aaccttctgt tcggtgatga aattatcact aatggttttc attcctgtga
     1441 aagtgatgag gaggatagag cctcacatgc aagctctagt gactggactc caaggccacg
     1501 gataggtcca tatacttttg ttcagcaaca tcttatgatt ggcacagatc ctcgaacaat
     1561 tcttaaagat ttattgccgg aaacaatacc tccacctgag ttggatgata tgacactgtg
     1621 gcagattgtt attaatatcc tttcagaacc accaaaaagg aaaaaaagaa aagatattaa
     1681 tacaattgaa gatgctgtga aattactgca agagtgcaaa aaaattatag ttctaactgg
     1741 agctggggtg tctgtttcat gtggaatacc tgacttcagg tcaagggatg gtatttatgc
     1801 tcgccttgct gtagacttcc cagatcttcc agatcctcaa gcgatgtttg atattgaata
     1861 tttcagaaaa gatccaagac cattcttcaa gtttgcaaag gaaatatatc ctggacaatt
     1921 ccagccatct ctctgtcaca aattcatagc cttgtcagat aaggaaggaa aactacttcg
     1981 caactatacc cagaacatag acacgctgga acaggttgcg ggaatccaaa ggataattca
     2041 gtgtcatggt tcctttgcaa cagcatcttg cctgatttgt aaatacaaag ttgactgtga
     2101 agctgtacga ggagatattt ttaatcaggt agttcctcga tgtcctaggt gcccagctga
     2161 tgaaccgctt gctatcatga aaccagagat tgtgtttttt ggtgaaaatt taccagaaca
     2221 gtttcataga gccatgaagt atgacaaaga tgaagttgac ctcctcattg ttattgggtc
     2281 ttccctcaaa gtaagaccag tagcactaat tccaagttcc ataccccatg aagtgcctca
     2341 gatattaatt aatagagaac ctttgcctca tctgcatttt gatgtagagc ttcttggaga
     2401 ctgtgatgtc ataattaatg aattgtgtca taggttaggt ggtgaatatg ccaaactttg
     2461 ctgtaaccct gtaaagcttt cagaaattac tgaaaaacct ccacgaacac aaaaagaatt
     2521 ggcttatttg tcagagttgc cacccacacc tcttcatgtt tcagaagact caagttcacc
     2581 agaaagaact tcaccaccag attcttcagt gattgtcaca cttttagacc aagcagctaa
     2641 gagtaatgat gatttagatg tgtctgaatc aaaaggttgt atggaagaaa aaccacagga
     2701 agtacaaact tctaggaatg ttgaaagtat tgctgaacag atggaaaatc cggatttgaa
     2761 gaatgttggt tctagtactg gggagaaaaa tgaaagaact tcagtggctg gaacagtgag
     2821 aaaatgctgg cctaatagag tggcaaagga gcagattagt aggcggcttg atggtaatca
     2881 gtatctgttt ttgccaccaa atcgttacat tttccatggc gctgaggtat attcagactc
     2941 tgaagatgac gtcttatcct ctagttcttg tggcagtaac agtgatagtg ggacatgcca
     3001 gagtccaagt ttagaagaac ccatggagga tgaaagtgaa attgaagaat tctacaatgg
     3061 cttagaagat gagcctgatg ttccagagag agctggagga gctggatttg ggactgatgg
     3121 agatgatcaa gaggcaatta atgaagctat atctgtgaaa caggaagtaa cagacatgaa
     3181 ctatccatca aacaaatcaa agggtgggcg cgccgaccca gctttcttgt acaaagtggt
     3241 tcgatctaga atggctagca aaggagaaga acttttcact ggagttgtcc caattcttgt
     3301 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
     3361 tgctacatac ggaaagctta cccttaaatt tatttgcact actggaaaac tacctgttcc
     3421 atggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttccc gttatccgga
     3481 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg
     3541 cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg
     3601 tgataccctt gttaatcgta tcgagttaaa aggtattgat tttaaagaag atggaaacat
     3661 tctcggacac aaactcgagt acaactataa ctcacacaat gtatacatca cggcagacaa
     3721 acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggatccgt
     3781 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     3841 agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga
     3901 ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct
     3961 ctacaaataa tgaattaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4021 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4081 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4141 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4201 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4261 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4321 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4381 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4441 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4501 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4561 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4621 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4681 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4861 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4921 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4981 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5041 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5101 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5161 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5221 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5281 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5341 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5401 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5461 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5521 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5581 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5641 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5701 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5761 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5821 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5881 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5941 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6001 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6061 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6181 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8401 gtc
//
