LOCUS       Exported                7104 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 7104)
  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..7104
                     /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..1900
                     /label=SIRT4
     protein_bind    complement(1934..1941)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1952..2671
                     /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    2699..2923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2969..3397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3411..3740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3591..3726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3807..4601
                     /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    4777..4898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5349..5937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6108..6968)
                     /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(6969..7073)
                     /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 gaagatgagc tttgcgttga ctttcaggtc agcaaaaggc cgttggatcg caaaccccag
     1021 ccagccgtgc tcgaaagcct ccattgggtt atttgtgcca gcaagtcctc ctctggaccc
     1081 tgagaaggtc aaagagttac agcgcttcat caccctttcc aagagactcc ttgtgatgac
     1141 tggggcagga atctccaccg aatcggggat accagactac aggtcagaaa aagtggggct
     1201 ttatgcccgc actgaccgca ggcccatcca gcatggtgat tttgtccgga gtgccccaat
     1261 ccgccagcgg tactgggcga gaaacttcgt aggctggcct caattctcct cccaccagcc
     1321 taaccctgca cactgggctt tgagcacctg ggagaaactc ggaaagctgt actggttggt
     1381 gacccaaaat gtggatgctt tgcacaccaa ggcggggagt cggcgcctga cagagctcca
     1441 cggatgcatg gacagggtcc tgtgcttgga ttgtggggaa cagactcccc ggggggtgct
     1501 gcaagagcgt ttccaagtcc tgaaccccac ctggagtgct gaggcccatg gcctggctcc
     1561 tgatggtgac gtctttctct cagaggagca agtccggagc tttcaggtcc caacctgcgt
     1621 tcaatgtgga ggccatctga aaccagatgt cgttttcttc ggggacacag tgaaccctga
     1681 caaggttgat tttgtgcaca agcgtgtaaa agaagccgac tccctcttgg tggtgggatc
     1741 atccttgcag gtatactctg gttacaggtt tatcctcact gcctgggaga agaagctccc
     1801 gattgcaata ctgaacattg ggcccacacg gtcggatgac ttggcgtgtc tgaaactgaa
     1861 ttctcgttgt ggagagttgc tgcctttgat agacccatgc aagggtgggc gcgccgaccc
     1921 agctttcttg tacaaagtgg ttcgatctag aatggctagc aaaggagaag aacttttcac
     1981 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     2041 cagtggagag ggtgaaggtg atgctacata cggaaagctt acccttaaat ttatttgcac
     2101 tactggaaaa ctacctgttc catggccaac acttgtcact actttctctt atggtgttca
     2161 atgcttttcc cgttatccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc
     2221 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg
     2281 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga
     2341 ttttaaagaa gatggaaaca ttctcggaca caaactcgag tacaactata actcacacaa
     2401 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca
     2461 caacattgaa gatggatccg ttcaactagc agaccattat caacaaaata ctccaattgg
     2521 cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa
     2581 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat
     2641 tacacatggc atggatgagc tctacaaata atgaattaaa cccgctgatc agcctcgact
     2701 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2761 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2821 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2881 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2941 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3001 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3061 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3121 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3181 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3241 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3301 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3361 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3421 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3661 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3721 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3781 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4621 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4681 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4741 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4801 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4861 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4921 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5761 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7081 ccccgaaaag tgccacctga cgtc
//
