LOCUS       Exported                7713 bp ds-DNA     circular SYN 29-MAR-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7713)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 29, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7713
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        29..458
                     /label=CUP1 promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             486..926
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /note="color: #993366"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     937..956
                     /label=T7
                     /note="color: black; sequence: TAATACGACTCACTATAGGG; added:
                     2021-03-29"
     promoter        937..955
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             975..1004
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             1008..1718
                     /codon_start=1
                     /label=mGrape3
                     /note="color: #800080"
                     /translation="MVSKGEENNMAVIKEFMRFKTHMEGSVNGHEFEIEGKGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQMMYGSKAYVKHPPDIPDYMKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLHGTNFPSDGPVMQKKTMGWEASSERLYPEDGALK
                     GEVKMRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKLDYKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      1748..1795
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     terminator      1822..2009
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
                     /note="color: #ffffff"
     primer_bind     complement(2033..2049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(2033..2049)
                     /label=M13R
                     /note="color: black; sequence: CAGGAAACAGCTATGAC; added: 
                     2021-03-29"
     protein_bind    2057..2073
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(2081..2111)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2081 .. 2087 / #ffffff / -10
                        2: 2088 .. 2105 / #ffffff
                        3: 2106 .. 2111 / #ffffff / -35"
     protein_bind    2126..2147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(2435..3023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     polyA_signal    3352..3399
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     CDS             complement(3631..4425)
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4426..4530)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4557..5899
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
                     /note="color: #ffff00"
     protein_bind    complement(5520..5567)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /note="color: #31849b; direction: LEFT"
     promoter        6158..6439
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             6440..7114
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /note="color: #ff7f50"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7217..7672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tcccattacc gacatttggg cgctatacgt
       61 gcataagttc atgtatgtat ctgtatttaa aacacttttg tattattttt cctcatatat
      121 gtgtataggt ttatacggat gatttaatta ttacttcacc accctttatt tcaggctgat
      181 atcttagcct tgttactagt tagaaaaaga catttttgct gtcagtcact gtcaagagat
      241 tcttttgctg gcatttcttc tagaagcaaa aagagcgatg cgtcttttcc gctgaaccgt
      301 tccagcaaaa aagactacca acgcaatatg gattgtcaga atcatataaa agagaagcaa
      361 ataactcctt gtcttgtatc aattgcatta taatatcttc ttgttagtgc aatatcatat
      421 agaagtcatc gaaatagata ttaagaaaaa caaactgtcc aagcttgaag caagcctcct
      481 gaaagatgaa gctactgtct tctatcgaac aagcatgcga tatttgccga cttaaaaagc
      541 tcaagtgctc caaagaaaaa ccgaagtgcg ccaagtgtct gaagaacaac tgggagtgtc
      601 gctactctcc caaaaccaaa aggtctccgc tgactagggc acatctgaca gaagtggaat
      661 caaggctaga aagactggaa cagctatttc tactgatttt tcctcgagaa gaccttgaca
      721 tgattttgaa aatggattct ttacaggata taaaagcatt gttaacagga ttatttgtac
      781 aagataatgt gaataaagat gccgtcacag atagattggc ttcagtggag actgatatgc
      841 ctctaacatt gagacagcat agaataagtg cgacatcatc atcggaagag agtagtaaca
      901 aaggtcaaag acagttgact gtatcgccgg aatttgtaat acgactcact atagggcgag
      961 ccgccatcat ggaggagcag aagctgatct cagaggagga cctgcatatg gtgagcaagg
     1021 gcgaggagaa taacatggcc gtcatcaagg agttcatgcg cttcaagacg cacatggagg
     1081 gctccgtgaa cggccacgag ttcgagatcg agggcaaggg cgagggccgc ccctacgagg
     1141 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     1201 tcctgtcccc tcagatgatg tacggctcca aggcctacgt gaagcacccc cctgacatcc
     1261 ccgactacat gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1321 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     1381 acaaggtgaa gctgcacggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     1441 ccatgggctg ggaggcctcc tccgagcggt tgtaccccga ggacggcgcc ctgaagggcg
     1501 aggtcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca
     1561 cctacatggc caagaagccc gtgcagctgc ccggcgccta caagctcgac tacaagctgg
     1621 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc
     1681 gccactccac cggcggcatg gacgagctgt acaagtaagg atccgtcgac ctgcagcggc
     1741 cgcataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgcgcgc
     1801 ttgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat tattaaataa
     1861 gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa
     1921 ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag
     1981 gtcgctctta ttgaccacac ctctaccggc atgcaagctt ggcgtaatca tggtcatagc
     2041 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2101 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac
     3181 ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg
     3241 aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc
     3301 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
     3361 acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     3421 gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca
     3481 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3541 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     3601 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     3661 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     3721 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     3781 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     3841 attcggcaag caggcatcgc catgagtcac gacgagatcc tcgccgtcgg gcatgctcgc
     3901 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     3961 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     4021 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     4081 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     4141 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     4201 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc
     4261 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     4321 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     4381 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatactct tcctttttca
     4441 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4501 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaacg
     4561 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     4621 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     4681 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     4741 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     4801 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     4861 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     4921 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     4981 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     5041 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     5101 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     5161 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     5221 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     5281 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     5341 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     5401 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     5461 gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg
     5521 aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga
     5581 aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg
     5641 cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg
     5701 tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta
     5761 cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct
     5821 ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca
     5881 tttcctttga tattggatca tactaagaaa ccattattat catgacatta acctataaaa
     5941 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6001 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6061 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6121 catcagagca gattgtactg agagtgcacc atagatcaac gacattacta tatatataat
     6181 ataggaagca tttaatagaa cagcatcgta atatatgtgt actttgcagt tatgacgcca
     6241 gatggcagta gtggaagata ttctttattg aaaaatagct tgtcacctta cgtacaatct
     6301 tgatccggag cttttctttt tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg
     6361 agagggccaa gagggagggc attggtgact attgagcacg tgagtatacg tgattaagca
     6421 cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg gtccattggt
     6481 gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt ccgatgctga
     6541 cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag
     6601 gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt
     6661 tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat
     6721 tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt tcctcggttt
     6781 gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc
     6841 acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga caggtgaact
     6901 tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg aaagcttaca
     6961 ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta gattaaatgg
     7021 cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa
     7081 aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt atttatttaa
     7141 gtattgtttg tgcacttgcc gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag
     7201 aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     7261 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     7321 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     7381 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     7441 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     7501 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     7561 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     7621 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
     7681 cattcaggct gcgcaactgt tgggaagggc gat
//
