LOCUS       Exported                7722 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 7722)
  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..7722
                     /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..1727
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1008 .. 1010 / #05fd14
                        2: 1011 .. 1013 / #05fd14 / 1a
                        3: 1014 .. 1727 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      1757..1804
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     terminator      1831..2018
                     /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(2042..2058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(2042..2058)
                     /label=M13R
                     /note="color: black; sequence: CAGGAAACAGCTATGAC; added: 
                     2021-03-29"
     protein_bind    2066..2082
                     /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(2090..2120)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2090 .. 2096 / #ffffff / -10
                        2: 2097 .. 2114 / #ffffff
                        3: 2115 .. 2120 / #ffffff / -35"
     protein_bind    2135..2156
                     /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(2444..3032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     polyA_signal    3361..3408
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     CDS             complement(3640..4434)
                     /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(4435..4539)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4566..5908
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
                     /note="color: #ffff00"
     protein_bind    complement(5529..5576)
                     /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        6167..6448
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             6449..7123
                     /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(7226..7681)
                     /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 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1081 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1141 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1201 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     1261 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1321 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1381 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1441 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1501 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1561 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1621 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1681 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagga tccgtcgacc
     1741 tgcagcggcc gcataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1801 tttgcgcgct tgcagccaag ctaattccgg gcgaatttct tatgatttat gatttttatt
     1861 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
     1921 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
     1981 tagcatgagg tcgctcttat tgaccacacc tctaccggca tgcaagcttg gcgtaatcat
     2041 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2881 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3181 atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc
     3241 cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc
     3301 aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat
     3361 gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg
     3421 ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg tgggcgaaga
     3481 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     3541 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     3601 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
     3661 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     3721 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     3781 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     3841 caccatgata ttcggcaagc aggcatcgcc atgagtcacg acgagatcct cgccgtcggg
     3901 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     3961 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     4021 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     4081 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     4141 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     4201 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc
     4261 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     4321 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     4381 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatactctt
     4441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4561 acctgaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     4621 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     4681 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     4741 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     4801 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     4861 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     4921 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     4981 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     5041 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     5101 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     5161 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     5221 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     5281 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     5341 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     5401 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     5461 cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     5521 agcgctctga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaag
     5581 cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg
     5641 ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca
     5701 tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt
     5761 agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag
     5821 cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca
     5881 atgctatcat ttcctttgat attggatcat actaagaaac cattattatc atgacattaa
     5941 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     6001 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6061 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6121 actatgcggc atcagagcag attgtactga gagtgcacca tagatcaacg acattactat
     6181 atatataata taggaagcat ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt
     6241 atgacgccag atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac
     6301 gtacaatctt gatccggagc ttttcttttt ttgccgatta agaattaatt cggtcgaaaa
     6361 aagaaaagga gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt
     6421 gattaagcac acaaaggcag cttggagtat gtctgttatt aatttcacag gtagttctgg
     6481 tccattggtg aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg ctctagattc
     6541 cgatgctgac ttgctgggta ttatatgtgt gcccaataga aagagaacaa ttgacccggt
     6601 tattgcaagg aaaatttcaa gtcttgtaaa agcatataaa aatagttcag gcactccgaa
     6661 atacttggtt ggcgtgtttc gtaatcaacc taaggaggat gttttggctc tggtcaatga
     6721 ttacggcatt gatatcgtcc aactgcatgg agatgagtcg tggcaagaat accaagagtt
     6781 cctcggtttg ccagttatta aaagactcgt atttccaaaa gactgcaaca tactactcag
     6841 tgcagcttca cagaaacctc attcgtttat tcccttgttt gattcagaag caggtgggac
     6901 aggtgaactt ttggattgga actcgatttc tgactgggtt ggaaggcaag agagccccga
     6961 aagcttacat tttatgttag ctggtggact gacgccagaa aatgttggtg atgcgcttag
     7021 attaaatggc gttattggtg ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga
     7081 ctctaacaaa atagcaaatt tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta
     7141 tttatttaag tattgtttgt gcacttgccg atctatgcgg tgtgaaatac cgcacagatg
     7201 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     7261 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     7321 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     7381 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7441 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     7501 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7561 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7621 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7681 tccattcgcc attcaggctg cgcaactgtt gggaagggcg at
//
