LOCUS       Exported                6272 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    pSW_1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6272)
  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 6272)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6272
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        359..738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        739..942
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     893..913
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     939..963
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1078..1210
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1193..1212
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1255..1273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1292..1333
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1343..1783
                     /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    1805..1929
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1954..1984
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2038..2718
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(2104..2123)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3038..3343
                     /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     3222..3241
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3384..3508)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          3600..3665
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3795..3815
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4240..4374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4277..4296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4331..4350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             4471..5331
                     /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(4689..4708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5502..6090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5991..6010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6244..6261
                     /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 ttcaatattg gccattagcc
      241 atattattca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
      301 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg
      361 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      421 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      481 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      541 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      601 gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      661 cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta
      721 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
      781 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      841 caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg
      901 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      961 atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct
     1021 tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac tgggcaggta
     1081 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     1141 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     1201 ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac
     1261 gactcactat aggctagcca ccgatatcat ggggaagcca atccctaatc cccttctggg
     1321 actcgactct accggcggct ctatgaagct actgtcttct atcgaacaag catgcgatat
     1381 ttgccgactt aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa
     1441 gaacaactgg gagtgtcgct actctcccaa aaccaaaagg tctccgctga ctagggcaca
     1501 tctgacagaa gtggaatcaa ggctagaaag actggaacag ctatttctac tgatttttcc
     1561 tcgagaagac cttgacatga ttttgaaaat ggattcttta caggatataa aagcattgtt
     1621 aacaggatta tttgtacaag ataatgtgaa taaagatgcc gtcacagata gattggcttc
     1681 agtggagact gatatgcctc taacattgag acagcataga ataagtgcga catcatcatc
     1741 ggaagagagt agtaacaaag gtcaaagaca gttgactgta tcgtcgggcg gcggccaccg
     1801 gtatacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
     1861 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     1921 gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1981 aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg
     2041 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     2101 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     2161 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     2221 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     2281 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     2341 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     2401 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     2461 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2521 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     2581 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2641 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcaggcggg gcgtaatcta
     2701 gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     2761 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
     2821 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     2881 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     2941 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     3001 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     3061 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     3121 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg
     3181 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     3241 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     3301 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     3361 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
     3421 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     3481 ctcgttcagc tttcttgtac aaagtggtga cgtaagctag caggatcttt gtgaaggaac
     3541 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg
     3601 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat
     3661 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg
     3721 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc
     3781 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag
     3841 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta
     3901 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg
     3961 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac
     4021 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt
     4081 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata
     4141 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4201 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4261 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4321 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcccgttt
     4381 aaactacgcg taattcaaac agggttctgg cgtcgttctc gtactgtttt ccccaggcca
     4441 gtgctttagc gttattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4501 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4561 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4621 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4681 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4741 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4801 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4861 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4921 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4981 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5041 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5101 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5161 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5221 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5281 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5341 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5401 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5461 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5521 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5581 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5641 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5701 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5761 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5821 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5881 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5941 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6001 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6061 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6121 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6181 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6241 cgcagcgagt cagtgagcga ggaagcggaa ga
//
