LOCUS       sequence-127115-        7649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7649)
  AUTHORS   Stampfel G, Kazmar T, Frank O, Wienerroither S, Reiter F, Stark A
  TITLE     Transcriptional regulators form diverse groups with 
            context-dependent regulatory functions.
  JOURNAL   Nature. 2015 Nov 9. doi: 10.1038/nature15545.
  PUBMED    26550828
REFERENCE   2  (bases 1 to 7649)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2015 Nov 9. doi: 10.1038/nature15545."
COMMENT     Sequence Label: pSGE_240_pAGW_V5
FEATURES             Location/Qualifiers
     source          1..7649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        140..2649
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2669..2710
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2720..3160
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    3182..3306
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3331..3336,3337..3354,3355..3361)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3415..4095
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(3481..3500)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4415..4720
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4599..4618
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4761..4885)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          4977..5042
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5172..5192
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5617..5751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5654..5673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5708..5727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(5848..5916,5917..6708)
                     /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"
     primer_bind     complement(6066..6085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6879..7467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7368..7387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7621..7638
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggcaact
       61 cgtgaaaggt aggcggatca gcggccgcag catgcaattc tatattctaa aaacacaaat
      121 gatacttcta aaaaaaaatc atgaatggca tcaactctga atcaaatctt tgcagatgca
      181 cctacttctc atttccactg tcacatcatt tttccagatc tcgctgcctg ttatgtggcc
      241 cacaaaccaa gacacgtttt atggccatta aagctggctg atcgtcgcca aacaccaaat
      301 acataatgaa tatgtacaca ttcgagaaag aagcgatcaa agaagcgtct tcgggcggag
      361 taggagaatg cggaggagaa ggagaacgag ctgatctagt atctctccac aatccaatgc
      421 caactgacca actggccata ttcggagcaa tttgaagcca atttccatcg cctggcgatc
      481 gctccattct tggctatatg tttttcaccg ttacccgggg ccattttcaa agactcgtcg
      541 gcaagataag attgtgtcac tcgctgtctc tcttcatttg tcgaagaatg ctgaggaatt
      601 tcgcgatgac gtcggcgagt attttgaaga atgagaataa tttgtattta tacgaaaatc
      661 agttagtgga attttctaca aaaacatgtt atctatagat aattttgttg caaaatatgt
      721 tgactatgac aaagattgta tgtatatacc tttaatgtat tctcattttc ttatgtattt
      781 ataatggcaa tgatgatact gatgatattt taagatgatg ccagaccaaa aggcttgaat
      841 ttctgcgtct tttgccgaac gcagtgcatg tgcaattgtt gttttttgga atattcaatt
      901 ttcggactgt ccgctttgat ttcagtttct tggcttattc aaaaagcaaa gtaaagccaa
      961 aaaagcgaga tggcaatacc aaatgcggca aaacggtagt ggaaggaaag gggtgcgggg
     1021 cagcggaagg aagggtgggg cggggcgtgg cggggtctgt ggctgggcgc gacgtcaccg
     1081 acgttggagc cactcctttg accatgtgtg cgtgtgtgta ttattcgtgt ctcgccactc
     1141 gccggttgtt tttttctttt tatgctgcgc tctctctagc gccatctcgc ttacgcatgc
     1201 tcaacgcacc gcatgttgcc gtttcctttt atgcgtcatt ttggctcgaa ataggcaatt
     1261 atttaaacaa agattagtca acgaaaacgc taaaataaat aagtctacaa tatggttact
     1321 tattgccatg tgtgtgcagc caacgatagc aacaaaagca acaacacagg tggctttccc
     1381 tctttcactt tttgtttgca agccgcgtgc gagcaagacg gcacgaccgg caaacgcaat
     1441 tacgctgaca aagagcagac gaagttttgg cgaaaaacat caaggcgcct gatacgaatg
     1501 catttgcaat aacaattgcg atatttaata ttgtttatga agctgtttga cttcaaaaca
     1561 cacaaaaaaa aaaataaaac aaattatttg aaagagaatt aggaatcgga cgcttatcgt
     1621 tagggtaaca acaagaaatg cttactgagt cacagcctct ggaaaactgc cgcaagccag
     1681 agagagagag aaaaagaggg agagcagctt agaccgcatg tgcttgtgtg tgaggcgtct
     1741 ctctcttcgt ctctgttgcg caaacgcata gactgcactg agaaaatcga ttacctattt
     1801 tttatgaatg aatatttgca ctattactat tcaaaactat taagatagca atcacattca
     1861 atagccaaat actataccac ctgagcgatg caacgaaatg atcaatttga gcaaaaatgc
     1921 tgcatattta ggacggcatc attatagaaa tgcttcttgc tgtgtacttt tctctcgtct
     1981 ggcagctgtt tcgccgttat tgttaaaacc ggcttaagtt aggtgtgttt tctacgacta
     2041 gtgaatgccc tactagaaga tgtgtgttgc acaaaatgtc cctggaataa ccaatttgaa
     2101 gtgcagatag cagtaaacgt aagctaatat gaatattatt taactgtaat gttttaatat
     2161 cgctggacat tactaataaa cccactataa acacatgtac atatgtatgt tttggcatac
     2221 aatgagtagt tggggaaaaa atgtgtaaaa gcaccgtgac catcacagca taaagataac
     2281 cagctgaagt atcgaatatg agtaaccccc aaattgaatc acatgccgca actgatagga
     2341 cccatggaag tacactcttc atggcgatat acaagacaca cacaagcacg aacacccagt
     2401 tgcggaggaa attctccgta aatgaaaacc caatcggcga acaattcata cccatatatg
     2461 gtaaaagttt tgaacgcgac ttgagagcgg agagcattgc ggctgataag gttttagcgc
     2521 taagcgggct ttataaaacg ggctgcggga ccagttttca tatcactacc gtttgagttc
     2581 ttgtgctgtg tggatactcc tcccgacaca aagccgctcc atcagccagc agtcgtctaa
     2641 tccagagacc ccggatccag atatcatggg gaagccaatc cctaatcccc ttctgggact
     2701 cgactctacc ggcggctcta tgaagctact gtcttctatc gaacaagcat gcgatatttg
     2761 ccgacttaaa aagctcaagt gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa
     2821 caactgggag tgtcgctact ctcccaaaac caaaaggtct ccgctgacta gggcacatct
     2881 gacagaagtg gaatcaaggc tagaaagact ggaacagcta tttctactga tttttcctcg
     2941 agaagacctt gacatgattt tgaaaatgga ttctttacag gatataaaag cattgttaac
     3001 aggattattt gtacaagata atgtgaataa agatgccgtc acagatagat tggcttcagt
     3061 ggagactgat atgcctctaa cattgagaca gcatagaata agtgcgacat catcatcgga
     3121 agagagtagt aacaaaggtc aaagacagtt gactgtatcg tcgggcggcg gccaccggta
     3181 tacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
     3241 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     3301 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     3361 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     3421 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     3481 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     3541 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     3601 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     3661 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     3721 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     3781 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     3841 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     3901 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     3961 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     4021 agaatgctta atgaattaca acagtactgc gatgagtggc aggcggggcg taatctagag
     4081 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     4141 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     4201 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     4261 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     4321 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     4381 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     4441 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     4501 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tcccccgtga
     4561 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     4621 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     4681 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     4741 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     4801 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     4861 gttcagcttt cttgtacaaa gtggtgacgt aagctagcag gatctttgtg aaggaacctt
     4921 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
     4981 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     5041 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
     5101 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
     5161 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
     5221 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
     5281 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa
     5341 cctttataag taggcataac agttataatc ataacatact gttttttctt actccacaca
     5401 ggcatagagt gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa
     5461 tttgtaaagg ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc
     5521 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     5581 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     5641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5701 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat cccgtttaaa
     5761 ctacgcgtaa ttcaaacagg gttctggcgt cgttctcgta ctgttttccc caggccagtg
     5821 ctttagcgtt attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5881 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5941 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6001 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6061 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6121 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6181 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6241 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6301 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6361 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6421 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6481 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6541 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6601 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6661 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6721 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6781 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6901 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6961 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7021 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7081 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7141 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7201 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7261 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7321 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7381 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7441 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7501 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7561 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7621 agcgagtcag tgagcgagga agcggaaga
//
